The Multitasker Protein: A Look at the Multiple Capabilities of NUMB.

NUMB, a plasma membrane-associated protein originally described in Drosophila, is involved in determining cell function and fate during early stages of development. It is secreted asymmetrically in dividing cells, with one daughter cell inheriting NUMB and the other inheriting its antagonist, NOTCH. NUMB has been proposed as a polarizing agent and has multiple functions, including endocytosis and serving as an adaptor in various cellular pathways such as NOTCH, Hedgehog, and the P53-MDM2 axis. Due to its role in maintaining cellular homeostasis, it has been suggested that NUMB may be involved in various human pathologies such as cancer and Alzheimer's disease. Further research on NUMB could aid in understanding disease mechanisms and advancing the field of personalized medicine and the development of new therapies.

IL-23 signaling is not an important driver of liver inflammation and fibrosis in murine non-alcoholic steatohepatitis models.

Non-alcoholic fatty liver disease (NAFLD), represents an unmet medical need that can progress to non-alcoholic steatohepatitis (NASH), which, without intervention, can result in the development of cirrhosis and hepatocellular carcinoma (HCC). Inflammation is a pathological hallmark of NASH, and targeting key inflammatory mediators of NASH may lead to potential therapeutics for the disease. Herein, we aimed to investigate the role of IL-23 signaling in NASH progression in murine models. We showed that recombinant IL-23 can promote IL-17 producing cell expansion in the liver and that these cells are predominately γδ T cells and Mucosal Associated Invariant T cells (MAITs). Reciprocally, we found that IL-23 signaling is necessary for the expansion of γδ T cells and MAIT cells in the western diet (WD) diet induced NASH model. However, we did not observe any significant differences in liver inflammation and fibrosis between wild type and Il23r-/- mice in the same NASH model. Furthermore, we found that Il23r deletion does not impact liver inflammation and fibrosis in the choline-deficient, L-amino acid-defined and high-fat diet (CDA-HFD) induced NASH model. Based on these findings, we therefore propose that IL-23 signaling is not necessary for NASH pathogenesis in preclinical models and targeting this pathway alone may not be an effective therapeutic approach to ameliorate the disease progression in NASH patients.

Biosíntesis inducida de fengicina y surfactina en una cepa de Bacillus amyloliquefaciens con actividad oomiceticida sobre zoosporas de Phytophthora capsica / Induced biosynthesis of fengycin and surfactin in a strain of Bacillus amyloliquefaciens with oomyceticidal activity on zoospores of Phytophthora capsici

Resumen La aplicación de metabolitos antimicrobianos biosintetizados por especies de Bacillus es una alternativa potencial para controlar Phytophthora capsici (P. capsici) en hortalizas y podría evitar el uso de productos químicos con acción oomiceticida. El objetivo de este estudio fue evaluar el impacto de la adición al medio de cultivo de distintos agentes (ácido glutámico, hierro, celulosa, quitina y células inactivas de Colletotrichum spp.) sobre la biosíntesis de lipopéptidos en Bacillus amyloliquefaciens KX953161.1 y examinar la capacidad oomiceticida de dichos compuestos in vitro sobre las zoosporas de P. capsici. Los lipopéptidos identificados y cuantificados por cromatografía en capa fina de alta resolución (HPTLC) en los extractos crudos fueron fengicina y surfactina. El cultivo bacteriano adicionado con células inactivas de Colletotrichum spp. demostró la mayor biosíntesis de lipopéptidos: 1.847,02 ±11,8 pg/mL de fengicina y 2.563,45 ± 18,4 pg/mL de surfactina. Los tratamientos con menor producción de estos lipopéptidos fueron aquellos a los que se añadió hierro (608,05 ± 22,6 pg/mL de fengicina y 903,74 ±22,1 pg/mL de surfactina) o celulosa (563,31 ±11,9 y 936,96 ±41,1 pg/mL, igual orden). El extracto con los lipopéptidos presentó una inhibición del 100% en la germinación de zoosporas de P. capsici, se observó enquistamiento, malformaciones en el tubo germinal y degradación celular. Se concluye que los lipopéptidos producidos por B. amyloliquefaciens KX953161.1 podrían contribuir al control de P. capsici, sin embargo, se requieren más estudios a fin de elucidar el modo de acción biológica de estos compuestos y optimizar el perfil de producción y el rendimiento.

Abstract A potential alternative to the use of chemical products with oomyceticidal action for the control of Phytophthora capsici in vegetables is the use of antimicrobial metabolites, biosynthesized in Bacillus species. The objective of this study was to induce the biosynthesis of lipopeptides in Bacillus amyloliquefaciens KX953161.1 by using glutamic acid, iron, cellulose, chitin, or inactive Colletotrichum spp. cells. The in vitro oomyceticidal effect of the bacterial lipopeptides on zoospores of Phytophthora capsici was evaluated. The lipopeptides identified and quantified in the crude extracts by high performance thin layer chromatography (HPTLC) were fengycin and surfactin. The bacterial culture with inactive fungal cells yielded the greatest biosynthesis of lipopeptides, at 1847.02± 11.8 and 2563.45± 18.4 pg/ml of fengycin and surfactin, respectively and the treatments that obtained lower production of these lipopepti-des, were those to which iron and cellulose were added with 608.05 ± 22.6 and 903.74± 22.1; 563.31± 11.9 and 936.96± 41.1 pg/ml for fengicin and surfactin, respectively. The lipopeptide extracted showed 100% germination inhibition on zoospores of P. capsici, revealing encystment, malformations in the germ tube and cellular degradation. Lipopeptides have the potential to control P. capsici; however, the biosynthesis of these lipopeptides requires further study to determine their biological mode of action and optimize lipopeptide performance and profile.

Comparison of the Effect of Hydrostatic and Dynamic High Pressure Processing on the Enzymatic Activity and Physicochemical Quality Attributes of 'Ataulfo' Mango Nectar.

The effects of hydrostatic (HHP) and dynamic (HPH) high-pressure treatments on the activity of pectin methylesterase (PME) and polyphenol oxidase (PPO) as well as the physicochemical quality attributes of 'Ataulfo' mango nectar were assessed. HHP reduced PME relative activity by 28% at 100 MPa for 5 min but increased PPO activity almost five-fold. Contrarily, HPH did not affect PME activity, but PPO was effectively reduced to 10% of residual activity at 300 MPa and at three passes. Color parameters (CIEL*a*b*), °hue, and chroma were differently affected by each type of high-pressure processing technology. The viscosity and fluid behavior were not affected by HHP, however, HPH changed the apparent viscosity at low dynamic pressure levels (100 MPa with one and three passes). The viscosity decreased at high shear rates in nectar samples, showing a shear-thinning effect. The results highlight how different effects can be achieved with each high-pressure technology; thus, selecting the most appropriate system for processing and preserving liquid foods like fruit beverages is recommended.

Effectiveness and persistence of golimumab as a second biological drug in patients with spondyloarthritis: A retrospective study.

ABSTRACT: This observational, longitudinal retrospective, noncomparative study was designed to assess the persistence and effectiveness of golimumab as a second anti-tumor necrosis factor (TNF) drug in patients with spondyloarthritis requiring discontinuation from a first anti-TNF drug.Data were collected retrospectively for all patients with axial spondyloarthritis or psoriatic arthritis from 20 rheumatology clinics in Spain who started golimumab as a second anti-TNF drug between January 2013 and December 2015. Golimumab persistence was assessed with Kaplan-Meier survival analysis, and associated factors were assessed with Cox regression analysis.210 patients started golimumab as a second anti-TNF drug: 131 with axial spondyloarthritis and 79 with psoriatic arthritis. In axial spondyloarthritis patients, the mean (standard deviation) Bath Ankylosing Spondylitis Disease Activity Index score at baseline was 5.5 (2.1), decreasing to 3.9 (2.0) at month 3 and 3.5 (2.0) at year 1, and remaining stable thereafter. In psoriatic arthritis patients, mean (standard deviation) baseline Disease Activity Score was 4.0 (1.3), reducing to 2.5 (1.2) at month 3 and to 2.2 (1.3) at year 1. Corresponding improvements were recorded from baseline in C-reactive protein levels and erythrocyte sedimentation rates. The probability of persistence of treatment with golimumab was 80% at year 1, 70% at year 2 and 65% at years 3 and year 4, and was similar in those who had stopped the first anti-TNF due to loss of efficacy or other reasons. Cox regression analysis showed that the probability of survival with golimumab was higher in patients with higher erythrocyte sedimentation rate, in patients with axial spondyloarthritis than with psoriatic arthritis, and in those who had discontinued adalimumab as first anti-TNF. Seventy-two patients (34.3%) discontinued golimumab during follow-up, 50 of them due to lack of efficacy.In patients with spondyloarthritis requiring discontinuation from a first anti-TNF drug, treatment with golimumab was effective and showed a high probability of persistence up to 4 years of treatment.

The peptide symporter SLC15a4 is essential for the development of systemic lupus erythematosus in murine models.

Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease representing a serious unmet medical need. The disease is associated with the loss of self-tolerance and exaggerated B cell activation, resulting in autoantibody production and the formation of immune complexes that accumulate in the kidney, causing glomerulonephritis. TLR7, an important mediator of the innate immune response, drives the expression of type-1 interferon (IFN), which leads to expression of type-1 IFN induced genes and aggravates lupus pathology. Because the lysosomal peptide symporter slc15a4 is critically required for type-1 interferon production by pDC, and for certain B cell functions in response to TLR7 and TLR9 signals, we considered it as a potential target for pharmacological intervention in SLE. We deleted the slc15a4 gene in C57BL/6, NZB, and NZW mice and found that pristane-challenged slc15a4-/- mice in the C57BL/6 background and lupus prone slc15a4-/- NZB/W F1 mice were both completely protected from lupus like disease. In the NZB/W F1 model, protection persisted even when disease development was accelerated with an adenovirus encoding IFNα, emphasizing a broad role of slc15a4 in disease initiation. Our results establish a non-redundant function of slc15a4 in regulating both innate and adaptive components of the immune response in SLE pathobiology and suggest that it may be an attractive drug target.

Autoencoded DNA methylation data to predict breast cancer recurrence: Machine learning models and gene-weight significance.

Breast cancer is the most frequent cancer in women and the second most frequent overall after lung cancer. Although the 5-year survival rate of breast cancer is relatively high, recurrence is also common which often involves metastasis with its consequent threat for patients. DNA methylation-derived databases have become an interesting primary source for supervised knowledge extraction regarding breast cancer. Unfortunately, the study of DNA methylation involves the processing of hundreds of thousands of features for every patient. DNA methylation is featured by High Dimension Low Sample Size which has shown well-known issues regarding feature selection and generation. Autoencoders (AEs) appear as a specific technique for conducting nonlinear feature fusion. Our main objective in this work is to design a procedure to summarize DNA methylation by taking advantage of AEs. Our proposal is able to generate new features from the values of CpG sites of patients with and without recurrence. Then, a limited set of relevant genes to characterize breast cancer recurrence is proposed by the application of survival analysis and a pondered ranking of genes according to the distribution of their CpG sites. To test our proposal we have selected a dataset from The Cancer Genome Atlas data portal and an AE with a single-hidden layer. The literature and enrichment analysis (based on genomic context and functional annotation) conducted regarding the genes obtained with our experiment confirmed that all of these genes were related to breast cancer recurrence.

Breast tumor cells promotes the horizontal propagation of EMT, stemness, and metastasis by transferring the MAP17 protein between subsets of neoplastic cells.

MAP17 (PDZK1IP1) is a small protein regulating inflammation and tumor progression, upregulated in a broad range of carcinomas. MAP17 levels increase during tumor progression in a large percentage of advanced tumors. In the present work, we explored the role of this protein shaping tumor evolution. Here we show that in breast cancer, cells increased MAP17 levels in tumors by demethylation induced multiple changes in gene expression through specific miRNAs downregulation. These miRNA changes are dependent on Notch pathway activation. As a consequence, epithelial mesenchymal transition (EMT) and stemness are induced promoting the metastatic potential of these cells both in vitro and in vivo. Furthermore, MAP17 increased the exosomes in tumor cells, where MAP17 was released as cargo, and this horizontal propagation also increased the EMT in the recipient cells. Importantly, an antibody against MAP17 in the media reduces the EMT and stemness alterations promoted by the conditioned media from MAP17-expressing cells. Therefore, MAP17 expression promotes the horizontal propagation of EMT and metastasis by transferring the MAP17 protein between subsets of neoplastic cells. Thus, MAP17 can be used to describe a new mechanism for cell malignity at distance, without the involvement of genetic or epigenetic modifications. MAP17 can also be taken in consideration as new target for metastatic high-grade breast tumors.

Sarcoma stratification by combined pH2AX and MAP17 (PDZK1IP1) levels for a better outcome on doxorubicin plus olaparib treatment.

Sarcomas constitute a rare heterogeneous group of tumors, including a wide variety of histological subtypes. Despite advances in our understanding of the pathophysiology of the disease, first-line sarcoma treatment options are still limited and new treatment approaches are needed. Histone H2AX phosphorylation is a sensitive marker for double strand breaks and has recently emerged as biomarker of DNA damage for new drug development. In this study, we explored the role of H2AX phosphorylation at Ser139 alone or in combination with MAP17 protein, an inducer of DNA damage through ROS increase, as prognostic biomarkers in sarcoma tumors. Next, we proposed doxorubicin and olaparib combination as potential therapeutic strategies against sarcomas displaying high level of both markers. We evaluate retrospectively the levels of pH2AX (Ser139) and MAP17 in a cohort of 69 patients with different sarcoma types and its relationship with clinical and pathological features. We found that the levels of pH2AX and MAP17 were related to clinical features and poor survival. Next, we pursued PARP1 inhibition with olaparib to potentiate the antitumor effect of DNA damaging effect of the DNA damaging agent doxorubicin to achieve an optimal synergy in sarcoma. We demonstrated that the combination of olaparib and doxorubicin was synergistic in vitro, inhibiting cell proliferation and enhancing pH2AX intranuclear accumulation, as a result of DNA damage. The synergism was corroborated in patient-derived xenografts (PDX) where the combination was effective in tumors with high levels of pH2AX and MAP17, suggesting that both biomarkers might potentially identify patients who better benefit from this combined therapy.

Role of Mitochondria in Cancer Stem Cell Resistance.

Cancer stem cells (CSC) are associated with the mechanisms of chemoresistance to different cytotoxic drugs or radiotherapy, as well as with tumor relapse and a poor prognosis. Various studies have shown that mitochondria play a central role in these processes because of the ability of this organelle to modify cell metabolism, allowing survival and avoiding apoptosis clearance of cancer cells. Thus, the whole mitochondrial cycle, from its biogenesis to its death, either by mitophagy or by apoptosis, can be targeted by different drugs to reduce mitochondrial fitness, allowing for a restored or increased sensitivity to chemotherapeutic drugs. Once mitochondrial misbalance is induced by a specific drug in any of the processes of mitochondrial metabolism, two elements are commonly boosted: an increment in reactive nitrogen/oxygen species and, subsequently, activation of the intrinsic apoptotic pathway.

Historias digitales mediante Windows Movie Maker, Una herramienta para mejorar la escritura en inglés / Digital stories through Windows Movie Maker: A tool to improve writing in English / Histórias digitais usando o Windows Movie Maker: uma ferramenta para melhorar a escrita em inglês

RESUMEN La escritura es una de las cuatro habilidades comunicativas del inglés que debe ser trabajada en clases; sin embargo, por su naturaleza, y al tratarse de un proceso, no puede desarrollarse apropiadamente en algunas ocasiones. Esta investigación-acción se realizó en un establecimiento educacional público de la región del Bio Bio - Chile. Se trabajó con I9 estudiantes de séptimo año de primaria y se implementó una intervención pedagógica que contribuyera a mejorar la habilidad de escritura en inglés de los estudiantes, mediante el desarrollo de historias digitales breves creadas con el programa Windows Movie Maker Se utilizó una tarea de escritura que fue evaluada con una rúbrica analítica, tanto al inicio como al final de la intervención. Para verificar la significancia de los resultados, se utilizó el test de Wilcoxon. Se aplicaron además cuestionarios de opiniones a los estudiantes para identificar sus percepciones respecto de esta intervención. Entre los resultados obtenidos, se puede destacar que la implementación de esta intervención no sólo contribuyó de manera significativa a la mejora en el desempeño de los estudiantes, al momento de escribir en inglés, sino que también ayudó a generar una sensación de agrado hacia las actividades de escritura en inglés.

ABSTRACT Writing is one of the four communicative skills that must be practiced inside the classroom; however; due to its nature and considering that writing is a process, it cannot be properly developed on certain occasions. This action research took place in a public school in the Bio Bio region - Chile. The participants of this study were I9 seventh grade students, and an intervention sequence was conducted to improve students' writing skill in English, by producing short digital stories using Windows Movie Maker. A writing task was developed by the students and assessed with an analytic rubric at the beginning and end of the intervention. The results were compared using the Wilcoxon test in order to determine if there were statistically significant differences. In addition, a perceptions questionnaire was applied to identify students' thoughts about the intervention. Regarding the results, it can be stated that this intervention not only helped to improve the students' performance when writing in English, but also contributed to generate a positive impression when they had to face writing tasks.

RESUMO Escrever é uma das quatro habilidades comunicativas do inglês que deve ser trabalhada em sala de aula; no entanto, devido à sua natureza, e como é um processo, ele não pode se desenvolver adequadamente em algumas ocasiões. Esta pesquisa-ação foi realizada em um estabelecimento educacional público na região de Bio Bio - Chile. Trabalhamos com I9 alunos do ensino fundamental da sétima série e uma intervenção pedagógica foi implementada para ajudar a melhorar a capacidade de escrita dos alunos em inglês, através do desenvolvimento de pequenas histórias digitais criadas com o programa Windows Movie Maker Foi utilizada uma tarefa de escrita avaliada com uma rubrica analítica, tanto no início quanto no final da intervenção. Para verificar a significancia dos resultados, foi utilizado o teste de Wilcoxon. Questionários de opinião também foram aplicados aos estudantes para identificar suas percepções em relação a essa intervenção. Dentre os resultados obtidos, destaca-se que a implementação dessa intervenção não apenas contribuiu significativamente para a melhoria do desempenho do aluno, no momento da redação em inglês, mas também contribuiu para gerar uma sensação de prazer em relação às atividades de escrever em inglês.

PAI1 is a Marker of Bad Prognosis in Rectal Cancer but Predicts a Better Response to Treatment with PIM Inhibitor AZD1208.

Colorectal cancer (CRC) is the third most common cancer worldwide. The standard treatment in locally advanced rectal cancer is preoperative radiation alone or in combination with chemotherapy, followed by adjuvant chemotherapy. Rectal cancer is highly lethal, with only 20% of patients showing a complete remission (by RECIST) after standard treatment, although they commonly show local or systemic relapse likely due to its late detection and high chemotherapy resistance, among other reasons. Here, we explored the role of PAI1 (Serpin E1) in rectal cancer through the analyses of public patient databases, our own cohort of locally advanced rectal cancer patients and a panel of CRC cell lines. We showed that PAI1 expression is upregulated in rectal tumors, which is associated with decreased overall survival and increased metastasis and invasion in advanced rectal tumors. Accordingly, PAI1 expression is correlated with the expression of (Epithelial-to-Mesenchymal Transition) EMT-associated genes and genes encoding drug targets, including the tyrosine kinases PDGFRb, PDGFRa and FYN, the serine/threonine kinase PIM1 and BRAF. In addition, we demonstrate that cells expressing PAI1 protein are more sensitive to the PIM inhibitor AZD1208, suggesting that PAI1 could be used to predict response to treatment with PIM inhibitors and to complement radiotherapy in rectal tumors.

Efecto de la temperatura y salinidad en la eclosión y supervivencia de Aedes aegypti (L) (Diptera: Culicidae) procedentes del occidente de México / Effect of temperature and salinity on the eclosion and survival of Aedes aegypti (L) (Diptera: Culicidae) from Western Mexico

Introducción: Aedes aegypti (L) (Diptera: Culicidae), es una especie cosmopolita y vector de arbovirosis. Las variaciones de la temperatura y salinidad del agua influyen en la eclosión y supervivencia de fases larvales. Objetivo: Evaluar el efecto de diferentes temperaturas y salinidades en la eclosión de huevos y la supervivencia de larvas, pupas y adultos bajo condiciones de laboratorio. Métodos: Se colectaron larvas de Ae. aegypti, de reservorios artificiales en la zona periurbana de Puerto Vallarta, Jalisco, México, y se mantuvieron hasta la fase adulta. Los huevos obtenidos se sometieron a ocho temperaturas (15, 17, 20, 25, 27, 30, 32 y 35 °C). Se colocaron 15 huevos por quintuplicado y se evaluó la eclosión durante 96 h. Se colocaron 100 huevos con agua ajustada a 0.3, 2, 5, 10, 15,18 y 22 ups y se evaluó la eclosión hasta las 96 h. Adicionalmente se utilizaron larvas del estadio IV, por quintuplicado, sometiéndose a las mismas salinidades y se evaluó la supervivencia hasta las 48 h. El efecto de la salinidad en la ovoposición de las hembras se llevó a cabo introduciendo recipientes con las mismas concentraciones salinas, dentro en las jaulas entomológicas. Resultados: Se registró el 100 por ciento de eclosión a las 24 y 48 h; la temperatura de 35° C no registró eclosión. Las salinidades de 22 y 18 ups, provocaron mortalidad del 100 por ciento a las 24 h. En la salinidad de 15 ups, sobrevivió el 50 por ciento. Las concentraciones de 2, 5 y 10 ups demostraron 100 por ciento de supervivencia hasta la fase de adulto. La supervivencia de larvas del estadio IV en los tratamientos 2, 5 y 10 fue del 100 por ciento y en 15,18 y 22 ups disminuyó a 50, 80 y 100 por ciento, respectivamente (p˂ 0,05). Las diferentes concentraciones salinas no afectaron significativamente la ovoposición. La eclosión solo se presentó en las concentraciones de 0,3; 2; 5 y 10 ups. Los huevos ovopositados en concentraciones de 15, 18 y 22 ups no eclosionaron hasta que fueron transferidos a agua dulce con porcentajes de eclosión de entre el 80 y 90 por ciento. Conclusiones: Los embriones de Ae. aegypti poseen una amplia plasticidad para soportar cambios drásticos de temperatura y salinidad. El control efectivo de sus poblaciones debe incluir la revisión de charcas o reservorios que contengan aguas salobres hasta 18 ups(AU)

Introduction: Aedes aegypti (L) (Diptera: Culicidae) is a cosmopolitan species and a vector of arboviruses. Variations in the temperature and salinity of the water affect eclosion and survival during the larval stages. Objective: Evaluate the effect of different temperatures and salinities on the eclosion of eggs and the survival of larvae, pupae and adults in laboratory conditions. Methods: Ae. aegypti larvae were collected from artificial reservoirs in a peri-urban area of Puerto Vallarta, Jalisco, Mexico, and maintained until the adult stage. The eggs obtained were subjected to eight temperatures (15, 17, 20, 25, 27, 30, 32 and 35 °C). Fifteen eggs were placed in quintuplicate and eclosion was evaluated for 96 h. One hundred eggs were placed with water adjusted to 0.3, 2, 5, 10, 15, 18 and 22 psu and eclosion was evaluated until 96 h. Additionally, stage IV larvae were used in quintuplicate, subjecting them to the same salinities and evaluating survival until 48 h. The effect of salinity on oviposition by females was determined by introducing containers with the same salinity into the entomological cages. Results: 100 percent eclosion was recorded at 24 and 48 h, whereas no eclosion occurred at a temperature of 35 °C. Salinities of 22 and 18 psu caused 100 percent mortality at 24 h, whereas 50 percent survived at a salinity of 15 psu. At concentrations of 2, 5 and 10 psu 100 percent of the larvae survived until the adult stage. Survival of stage IV larvae in treatments 2, 5 and 10 was 100%, whereas in 15, 18 and 22 psu it fell to 50, 80 and 100 percent, respectively (p˂ 0.05). The different salinities did not affect oviposition significantly. Eclosion only occurred at concentrations of 0.3, 2, 5 and 10 psu. Oviposited eggs at concentrations of 15, 18 and 22 psu did not eclose until they were transferred to fresh water, where eclosion percentages ranged between 80 percent and 90 percent. Conclusions: Ae. aegypti embryos have great plasticity to endure drastic changes in temperature and salinity. Effective control of their populations should include inspection of ponds and reservoirs containing brackish water of up to 18 psu(AU)

New markers for human ovarian cancer that link platinum resistance to the cancer stem cell phenotype and define new therapeutic combinations and diagnostic tools.

BACKGROUND: Ovarian cancer is the leading cause of gynecologic cancer-related death, due in part to a late diagnosis and a high rate of recurrence. Primary and acquired platinum resistance is related to a low response probability to subsequent lines of treatment and to a poor survival. Therefore, a comprehensive understanding of the mechanisms that drive platinum resistance is urgently needed. METHODS: We used bioinformatics analysis of public databases and RT-qPCR to quantitate the relative gene expression profiles of ovarian tumors. Many of the dysregulated genes were cancer stem cell (CSC) factors, and we analyzed its relation to therapeutic resistance in human primary tumors. We also performed clustering and in vitro analyses of therapy cytotoxicity in tumorspheres. RESULTS: Using bioinformatics analysis, we identified transcriptional targets that are common endpoints of genetic alterations linked to platinum resistance in ovarian tumors. Most of these genes are grouped into 4 main clusters related to the CSC phenotype, including the DNA damage, Notch and C-KIT/MAPK/MEK pathways. The relative expression of these genes, either alone or in combination, is related to prognosis and provide a connection between platinum resistance and the CSC phenotype. However, the expression of the CSC-related markers was heterogeneous in the resistant tumors, most likely because there were different CSC pools. Furthermore, our in vitro results showed that the inhibition of the CSC-related targets lying at the intersection of the DNA damage, Notch and C-KIT/MAPK/MEK pathways sensitize CSC-enriched tumorspheres to platinum therapies, suggesting a new option for the treatment of patients with platinum-resistant ovarian cancer. CONCLUSIONS: The current study presents a new approach to target the physiology of resistant ovarian tumor cells through the identification of core biomarkers. We hypothesize that the identified mutations confer platinum resistance by converging to activate a few pathways and to induce the expression of a few common, measurable and targetable essential genes. These pathways include the DNA damage, Notch and C-KIT/MAPK/MEK pathways. Finally, the combined inhibition of one of these pathways with platinum treatment increases the sensitivity of CSC-enriched tumorspheres to low doses of platinum, suggesting a new treatment for ovarian cancer.

Cellular antioxidant activity and in vitro intestinal permeability of phenolic compounds from four varieties of mango bark (Mangifera indica L.).

BACKGROUND: Mango bark is an important agro-industrial residue from mango pruning. In traditional medicine, the aqueous extract from mango bark (MBE) has been used in ethnomedicine for the treatment of many diseases. However, there is scarce information using cellular models to evaluate the potential use of this plant material for human consumption. In this study, the phenolic content from the MBE from four varieties (Kent, Keitt, Ataulfo and Tommy Atkins) was analyzed by high-performance liquid chromatography coupled to photodiode array detector (HPLC-DAD) and liquid chromatography coupled with time-of-flight mass spectrometry (LC/MS-TOF). Additionally, the cellular antioxidant activity of the MBE from the four mango varieties were compared. Finally, the intestinal permeability of the main polyphenols found in the MBE (mangiferin and gallic acid) was evaluated. RESULTS: Mangiferin and gallic acid were the main constituents in the MBE from the four mango varieties. Furthermore, the Ataulfo variety showed the highest cellular antioxidant activity (67%) at the concentration of 100 µg mL−1 . The intestinal permeability of mangiferin present in the bark extracts was 3- to 4.8-fold higher than those of mangiferin as standard, whereas the intestinal permeability of gallic acid varied among the tested extracts. CONCLUSION: MBE has the potential to exert antioxidant activity at the cellular level and can have an impact on human health. It may also be a good source for the extraction of polyphenols mainly mangiferin.

Skeletal Muscle Tissue Trib3 Links Obesity with Insulin Resistance by Autophagic Degradation of AKT2.

BACKGROUND/AIMS: Obesity is a serious health risk factor strongly associated with insulin resistance and type 2 diabetes; however, the underlying mechanisms associating obesity with insulin resistance remain unknown. In this study, we explored the physiological role of Trib3 in regulating glucose metabolism in skeletal muscle tissues in a Trib3 transgenic mice model. METHODS: Glucose metabolism in transgenic mice overexpressing Trib3 specifically in the skeletal muscle was examined by glucose/insulin tolerance test, metabolic cage studies, and glucose uptake assay. The effect of Trib3 overexpression on AKT phosphorylation and AKT protein turnover were assessed by RT-PCR and immunoblot analysis. Subcellular distribution of Trib3 and AKT1/2 was determined by microscopic analysis, co-immunoprecipitation experiments, and limited-detergent extraction of subcellular organelles. Ubiquitin assay was performed and ATG7 deficient cell line was employed to address the mechanisms of Trib3-dependent AKT protein homeostasis. RESULTS: We found that Trib3 expression in skeletal muscle is elevated in obese conditions, and transgenic mice that overexpressed Trib3, specifically in skeletal muscle tissues, displayed impaired glucose homeostasis by suppressing insulin-stimulated glucose uptake. Disruption of insulin signaling in skeletal muscle Trib3 transgenic mice may occur due to the specific downregulation of AKT2 but not AKT1. Autophagy regulated AKT2 protein turnover, and Trib3 overexpression stimulated autophagic degradation of AKT2 by promoting AKT2 ubiquitination. CONCLUSION: Because diet-induced obesity upregulates Trib3 and downregulates AKT2 in skeletal muscle tissues, Trib3 may play a key role in establishing an association between obesity and insulin resistance by regulating AKT2 protein homeostasis.

MAP17 predicts sensitivity to platinum-based therapy, EGFR inhibitors and the proteasome inhibitor bortezomib in lung adenocarcinoma.

BACKGROUND: The high incidence and mortality of lung tumours is a major health problem. Therefore, the identification both of biomarkers predicting efficacy for therapies in use and of novel efficacious therapeutic agents is crucial to increase patient survival. MAP17 (PDZK1IP1) is a small membrane-bound protein whose upregulation is reported as a common feature in tumours from diverse histological origins. Furthermore, MAP17 is correlated with tumour progression. METHODS: We assessed the expression of MAP17 in preclinical models, including cell lines and patient-derived xenografts (PDXs), assessing its correlation with sensitivity to different standard-of-care drugs in lung adenocarcinoma, as well as novel drugs. At the clinical level, we subsequently correlated MAP17 expression in human tumours with patient response to these therapies. RESULTS: We show that MAP17 expression is induced during lung tumourigenesis, particularly in lung adenocarcinomas, and provide in vitro and in vivo evidence that MAP17 levels predict sensitivity to therapies currently under clinical use in adenocarcinoma tumours, including cisplatin, carboplatin and EGFR inhibitors. In addition, we show that MAP17 expression predicts proteasome inhibitor efficacy in this context and that bortezomib, an FDA-approved drug, may be a novel therapeutic approach for MAP17-overexpressing lung adenocarcinomas. CONCLUSIONS: Our results indicate a potential prognostic role for MAP17 in lung tumours, with particular relevance in lung adenocarcinomas, and highlight the predictive pot0065ntial of this membrane-associated protein for platinum-based therapy and EGFR inhibitor efficacy. Furthermore, we propose bortezomib treatment as a novel and efficacious therapy for lung adenocarcinomas exhibiting high MAP17 expression.

Propiedades antioxidantes e inmunoestimulantes de polifenoles en peces carnívoros de cultivo / Antioxidant and immunostimulant properties of polyphenols in carnivorous farmed fish

Resumen El cultivo intensivo de peces es una estrategia econòmicamente importante para producir alimento. Sin embargo, las prácticas de cultivo intensivo generan estrés oxidativo e inmunosupresión, lo que ocasiona pérdidas de la calidad del especimen y aumento en la mortalidad. Para contrarrestar estos efectos, se ha optado por la administración de vegetales como fuente de polifenoles con propiedades antioxidantes e inmunoestimulantes en peces carnívoros de cultivo. El objetivo de este trabajo fue describir los efectos de los polifenoles de origen vegetal como antioxidantes e inmunoestimulantes en peces carnívoros, y promover su uso como ingredientes funcionales en la acuicultura. Los vegetales como fuente de polifenoles tienen la capacidad de mejorar los sistemas de defensa inmune y antioxidante de las especies analizadas, con un tejido de mejor calidad nutricional y un mayor contenido endógeno de antioxidantes. No obstante, las propiedades biológicas de los polifenoles dependen del tipo y concentra ción en el vegetal, de la dosis y el tiempo de administración, así como de la matriz alimentaria, la cual determina la bioaccesibilidad y biodisponibilidad de los polifenoles en el organismo. Es la información generada sobre el efecto de los polifenoles en la calidad post mortem, por lo que se deben realizar más estudios.

Abstract Fish production by intensive aquaculture, is an economically important strategy to produce food. However, intensive fish farming generates oxidative stress and suppress the immune system, causing loss of product quality and increasing fish mortality rates. To diminish these effects, plants as a source of polyphenols with antioxidants and immunostimulant properties were administered to carnivorous farmed fish. The aim of this study was to describe the effects of plant polyphenols as antioxidants and immunostimulants on carnivorous fish, and to promote their use as functional ingredients in aquaculture. Plants as a source of polyphenols showed the ability to improve the immune and antioxidant defense systems of the analyzed species, resulting in a tissue of better nutritional quality and a higher endogenous antioxidant content. However, the biological properties of polyphenols are dependent on the type of plant and their concentration within it, the dose and the time of administration, as well as the food matrix, which determines their bioaccessibility and bioavailability in the organism. There is little information on the effec of polyphenols in post mortem quality; therefore, further studies should be conducted.