IMCI approach in Tertiary hospitals, India

Objective: This study was conducted to compare physicians' diagnosis with Integrated Management of Childhood Illness (IMCI) algorithm generated diagnosis in hospitalized children aged 2-59 months. Methods: Recruited were patients aged 2-59 months admitted with one or more IMCI danger signs. IMCI and physician's diagnosis were noted and compared. Results: In 222 included subjects, mean duration of illness was 9.4 (SD: 16.5) days. Among those with cough or difficult breathing, 44 (19.8%) and 66 (29.7%) were diagnosed as either severe pneumonia or mild to moderate pneumonia by physicians and IMCI algorithm, respectively (p= 0.015). Among 146 presenting as fever, 140 (95.9%) were diagnosed as very severe febrile disease by the IMCI algorithm, whereas physicians diagnosed these as either malaria in 10/146 (6.7%), pyogenic meningitis in 47/146 (32.2%), sepsis in 31/146 (21.3%), tuberculous meningitis in 17/146 (11.6%), encephalitis in 5/146 (3.4%), measles in 3/146 (2.1%) or others in 24/146 (16.4%). Conclusion: As there was a low concordance between physician and IMCI algorithmic diagnosis of pneumonia (Kappa value= 0.74, 95% CI: (0.64-0.84)) and since very severe febrile disease is not a diagnosis made by the physicians, the IMCI algorithms have to be refined for appropriate management of these conditions

Programa de resolución de conflictos FHaCE up! Para la mejora de la convivencia y de la percepción de violencia en los centros de Educación Secundaria

La violencia escolar representa una de las problemáticas más graves a las que se enfrenta la comunidad educativa, debido a su alta prevalencia y las graves consecuencias asociadas, que pueden repercutir en el desempeño académico, la salud mental y la participación en actos delictivos y/o criminales, entre otras (Estévez et al., 2019; Gaffney et al., 2021; Polanin et al., 2021). Según el último informe proporcionado por Fundación ANAR, en España se archivaron un total de 2868 consultas en relación con episodios de violencia escolar, de las cuales 1911 casos fueron diagnosticados como acoso escolar (Ministerio de Educación y Formación Profesional, 2022b). Debido a la alta preocupación que genera abordar este fenómeno, existe una gran variedad de estudios que tienen como objetivo tratar y/o prevenir la violencia escolar, en concreto el acoso escolar tradicional y el ciberacoso. Diversos estudios defienden el abordaje del clima escolar para la prevención de la violencia (Bottiani et al., 2020; Bradshaw et al., 2022; Coyle et al., 2022), dado que juega un papel clave para el mantenimiento de relaciones de calidad entre los miembros de la comunidad educativa. En este sentido, la gestión pacífica de los conflictos, que surgen inevitablemente en el día a día de las escuelas, resulta una aproximación necesaria en materia de prevención de la violencia y la mejora del clima escolar (Chacon & Raj, 2022; Kafel, 2020; Turk, 2017). Dado que, evitar su afrontamiento o, lo que es peor, haciendo uso de la violencia, sea esta verbal, psicológica o física, puede agravar la situación, dando como resultado escuelas menos seguras (Wang & Degol, 2016). En materia de prevención de la violencia y gestión del clima escolar, muchos de los programas, que persiguen tales fines (Bonell et al., 2018; Ferrer-Cascales et al., 2019; Levi-Keren et al., 2022), desarrollan estrategias tanto de gestión de conflictos como de formación en competencias socioemocionales, incluidas también en el presente programa. Sin embargo, aun existiendo un consenso en cuanto a qué competencias predicen mejor la reducción de la violencia y la mejora del clima, como es el caso de la empatía (Garandeau et al., 2014; Montero-Carretero et al., 2021), no existe un consenso que defina cómo entrenar los estilos de afrontamiento del conflicto más adecuados para abordar la resolución pacífica de los mismos (Burger, 2022b; Rahim & Katz, 2019). Por ello, en el presente estudio, se decidió diseñar un programa de intervención educativa en resolución de conflictos, que desarrollara las competencias socioemocionales y comunicativas básicas, enfocadas a la configuración del estilo cooperativo, entendido como el estilo más adecuado, ya que muestra una gran preocupación tanto por la defensa de los propios intereses como de los intereses de los demás (DeVoogd et al., 2016; Kimsey & Fuller, 2003; Sánchez-Herrera et al., 2022). Es por todo ello, que el objetivo general del que parte el presente trabajo es diseñar y testear la efectividad de un programa de intervención para reducir la violencia y mejorar el clima escolar, a través de estrategias enfocadas en la resolución de conflictos de forma cooperativa. Para dicho objetivo se plantearon los siguientes objetivos específicos: 1) diseñar un programa de resolución de conflictos para la reducción de la violencia y la mejora del clima escolar; 2) validar una escala de escucha activa en población adolescente; 3) identificar los niveles de las variables objeto de estudio: empatía, habilidades sociales, escucha activa, estilos de afrontamiento del conflicto, clima y violencia escolar; 4) analizar la efectividad del programa de intervención; 5) analizar la relación entre las variables objeto de estudio: empatía, habilidades sociales, escucha activa, estilos de afrontamiento del conflicto, clima y violencia escolar. La investigación siguió un método cuasiexperimental pretest-postest y cumplió con todos los criterios éticos. El programa, llamado FHaCE up! está diseñado para trabajar con todo el alumnado de 1º a 4º de la ESO, durante el horario de tutoría, y está basado en diversos enfoques, comenzando con los principios de la teoría del aprendizaje social de Bandura (1998), el modelo de resolución de conflictos de Davidson y Woods (2004), el modelo de mediación entre iguales de Torrego (2013) y el modelo de inteligencia emocional de Salovey y Mayer (1990). Para poder llevar a cabo la propuesta, se contactó con dos centros públicos de la provincia de Valencia similares en sus características, los cuales se asignaron aleatoriamente a grupo control y grupo experimental. La muestra final estuvo compuesta por 264 estudiantes asignados a grupo experimental (55.3% mujeres) y 294 a grupo control (52.9% hombres), con edades comprendidas entre los 11 y los 17 años, grupo experimental (M=13.75, DT= 1.32) y grupo control (M= 13.84, DT=1.12). El programa se compuso de una doble formación: 1) competencias socioemocionales y comunicativas enfocadas a la resolución cooperativa de los conflictos; 2) implementación de un programa de mediación escolar entre iguales. Esta última con la finalidad de internalizar el estilo cooperativo. La duración total del programa se realizó a lo largo de todo el año académico 2018-2019. Tras los análisis llevados a cabo, se observaron resultados favorables para todos los objetivos planteados. Con respecto al primer objetivo, el programa en resolución de conflictos se llevó a cabo en un centro de educación secundaria de la provincia de Valencia, tras haberse recogido los consentimientos informados de las familias y del centro. En relación con al segundo objetivo, la validación de la escala de escucha activa cumplió con los criterios de bondad de ajuste y fiabilidad (α=.84). Los niveles de las variables de estudio delimitadas en el tercer objetivo presentaron valores medios o valores por encima del punto medio en el caso de las dimensiones de empatía, habilidades sociales, escucha activa, estilo cooperativo y clima escolar, y por debajo del punto medio en el caso de las dimensiones de violencia escolar, estilo agresivo y estilo evitativo. En cuanto al cuarto objetivo, las dimensiones que presentaron mayor efectividad del programa, con valores de η² que oscilaron entre .11 y .26, fueron empatía cognitiva y afectiva, escucha activa, expresar sentimientos positivos, rechazar peticiones, vínculos, violencia verbal de alumnado hacia alumnado, violencia física directa y amenazas entre estudiantes, y violencia a través de las NTIC. Por último, con respecto al quinto objetivo, las competencias socioemocionales desarrolladas en la intervención predijeron un 30% de la varianza del estilo cooperativo para el afrontamiento del conflicto en un sentido positivo, así como también esas variables predijeron entre un 12 y 14% las dimensiones claridad de normas y valores y vínculos de la variable clima escolar en sentido positivo, y entre un 6 y un 11% las dimensiones de violencia escolar en sentido negativo. Del estudio se concluyó que, el programa en resolución de conflictos FHaCE up! puede resultar una herramienta plausible para trabajar la mejora del clima y la reducción de la violencia escolar, aspectos esenciales para considerar una convivencia positiva En este sentido, la formación en competencias socioemocionales enfocadas a la gestión cooperativa de los conflictos, propuesta en la primera fase de formación, puede potenciar los beneficios de la implementación de un programa de mediación escolar entre iguales (Mas-Expósito et al., 2022; Seo & Kruis, 2022). Dado que, el estilo cooperativo requiere de una gran preocupación tanto por los propios intereses como por los intereses de los demás (DeVoogd et al., 2016; Kimsey & Fuller, 2003; Sánchez-Herrera et al., 2022), resulta el estilo ideal para tomar conciencia del daño ocasionado y aportar soluciones para repararlo durante el proceso de mediación (Burger, 2022b; Kafel, 2020; Rahim y Katz, 2019). Por otro lado, la mediación escolar entre iguales, que alude a los principios de autonomía y autoeficacia en la toma de decisiones durante la resolución de conflictos (Santamaría et al., 2021), fomenta la participación democrática (González et al., 2018; Cohen, 2008) y, esto a su vez, aumenta la seguridad, el apoyo y el compromiso del alumnado con su escuela, generando dinámicas de interacción más saludables, que mantengan un clima escolar positivo, un factor preventivo fundamental en la prevención de la violencia escolar (Abraczinskas et al., 2022; Crichlow-Ball & Cornell, 2021; Más-Expósito et al., 2022).School violence represents one of the most serious problems faced by the educational community, due to its high prevalence and the seriousness of its consequences, which can affect academic performance, mental health and participation in criminal acts and/or criminals among others (Estévez et al., 2019; Gaffney et al., 2021; Polanin et al., 2021). According to the latest report provided by Fundación ANAR, in Spain a total of 2,868 consultations were archived in relation to episodes of school violence, of which 1,911 cases were diagnosed as cases of bullying (Ministerio de Educación y Formación Profesional, 2022b). Due to the great concern generated by addressing this phenomenon, there is a wide variety of studies that aim to treat and/or prevent school violence, specifically traditional school bullying and cyberbullying. Various studies indicate that the school climate approach for violence prevention (Bottiani et al., 2020; Bradshaw et al., 2022; Coyle et al., 2022) since it emphasizes in the maintenance of quality relationships between members of the educational community as support and protection systems against violence. In this sense, the peaceful management of conflicts, which inevitably arise in the day-to-day life of schools, is a necessary approach in terms of violence prevention and improvement of the school climate (Chacon & Raj, 2022; Kafel, 2020; Turk, 2017). Given that avoiding confrontation or, what is worse, using violence, be it verbal, psychological or physical, can aggravate the situation, resulting in less safe schools (Wang & Degol, 2016). In terms of violence prevention and school climate management, many of the programs that pursue such purposes (Bonell et al., 2018; Ferrer-Cascales et al., 2019; Levi-Keren et al., 2022), develop strategies for both conflict management and training in socio-emotional skills, also included in this program. However, even though there is a consensus as to which competencies best predict the reduction of violence and the improvement of the climate, as is the case of empathy (Garandeau et al., 2014; Montero-Carretero et al., 2021), there is no consensus that defines how and which conflict coping styles to train in order to approach the peaceful resolution of conflicts (Burger, 2022b; Kimsey & Fuller, 2003; Rahim & Katz, 2019). For this reason, in the present study, it was decided to design an educational intervention program in conflict resolution, which would develop the basic socio-emotional and communicative skills focused on the configuration of the cooperative style, understood as the style that shows a great concern both for the defense both their own interests and the interests of others (DeVoogd et al., 2016; Sánchez-Herrera et al., 2022). This study aims to reduce violence and improve school climate through strategies focused on collaborative conflict management. For this purpose, the following specific objectives were proposed: 1) design a conflict resolution program; 2) to validate an active listening scale in the adolescent population; 3) identify the levels of the variables that are the object of the program; 4) to measure its effectiveness; and 5) analyze the relationship between variables, as well as their prediction. The variables developed in the program were: empathy, social skills, active listening, conflict management styles, climate and school violence. The research followed a quasi-experimental pretest-posttest method and met all the ethical criteria. The program called FHaCE up! it is designed to work with all students from 1st to 4th ESO, during tutoring hours, and is based on various approaches, beginning with the principles of Bandura's (1998) social learning theory, the problem-solving model conflicts of Davidson and Woods (2004), the peer mediation model of Torrego (2013) and the emotional intelligence model of Salovey and Mayer (1990). In order to carry out the proposal, two public secondary schools in the province of Valencia were contacted, which were randomly assigned to a control group and an experimental group. The final sample consisted of 264 students assigned to the experimental group (55.3% women) and 294 to the control group (52.9% men), aged between 11 and 17, experimental group (M=13.75, SD= 1.32) and control group (M= 13.84, SD=1.12). The program consisted of a double training: 1) socio-emotional and communication skills focused on the collaborative management style; 2) implementation of a peer-to-peer school mediation program. The latter in order to internalize the collaborative style. The total duration of the program was carried out throughout the entire 2018-2019 academic year. After the analysis carried out, favorable results were observed for all the proposed objectives. Regarding the first objective, the conflict resolution program was carried out in a secondary education center in the province of Valencia, after having obtained the informed consents of the families and the center. Regarding the second objective, the validation of the active listening scale met the criteria of goodness of fit and reliability (α=.84). The levels of the study variables delimited in the third objective presented mean values or values above the midpoint in the case of the dimensions of empathy, social skills, active listening, cooperative style, and school climate, and below the midpoint in the case of the dimensions of school violence, aggressive style and avoidant style. Regarding the fourth objective, the dimensions that presented the greatest effectiveness of the program, with values of η² that oscillated between .11 and .26, were cognitive and affective empathy, active listening, expressing positive feelings, rejecting requests, ties, verbal violence of students towards students, direct physical violence and threats between students, and violence through NICTs. Finally, with respect to the fifth objective, the socioemotional competencies developed in the intervention predicted 30% of the variance of the cooperative style for coping with conflict in a positive sense, as well as these variables predicted between 12 and 14% of the dimensions. clarity of norms and values and links of the school climate variable in a positive sense, and between 6 and 11% the dimensions of school violence in a negative sense. From the study it was concluded that the conflict resolution program FHaCE up! It can be a plausible tool to work on improving the climate and reducing school violence, essential aspects to consider a positive coexistence. In this sense, training in socio-emotional skills focused on the cooperative management of conflicts, proposed in the first phase of Training can enhance the benefits of implementing a school mediation program among peers (Mas-Expósito et al., 2022; Seo & Kruis, 2022). Given that the cooperative style requires great concern for both one's own interests and the interests of others (DeVoogd et al., 2016; Kimsey & Fuller, 2003; Sánchez-Herrera et al., 2022), the style is ideal for becoming aware of the damage caused and providing solutions to repair it during the mediation process (Burger, 2022b; Kafel, 2020; Rahim and Katz, 2019). On the other hand, school mediation between peers, which alludes to the principles of autonomy and self-efficacy in decision-making during conflict resolution (Santamaría et al., 2021), encourages democratic participation (González et al., 2018; Cohen, 2008) and this, in turn, increases the security, support and commitment of students to their school, generating healthier interaction dynamics that maintain a positive school climate, a fundamental preventive factor in the prevention of violence. school (Abraczinskas et al., 2022; Crichlow-Ball & Cornell, 2021; Más-Expósito et al., 2022)

Short communication: Laboratory evaluation of nine highbush blueberry cultivars susceptibility to Drosophila suzukii (Matsumura, 1931) in the Southwestern Spain

Aim of study:  To determine how susceptible the most used Southern highbush blueberry (SHB) cultivars were to the spotted wing Drosophila (SWD), Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae) as well as those recently introduced to Southwestern Spain.Area of study: Southwestern Spain (Huelva province).Material and methods: Nine of the SHB cultivars which were recently introduced in Southwestern Spain and the most used ones were selected: ‘Arana’, ‘Camellia’, ‘Kirra’, ‘Mayra’, ‘Misty’, ‘O'Neal’, ‘Sharpblue’, ‘Star’ and ‘Ventura’. In order to determine how susceptible the cultivars were to SWD, no-choice tests were performed under laboratory conditions. In addition, berry size, berry firmness, ºBrix, and pH were recorded in order to assess what influence these variables had on oviposition preference by SWD.Main results: Mean clutch size and mean number of emerged adults in ‘Star’ were significantly higher than in the other tested cultivars. ‘Mayra’, ‘Camellia’ and ‘Ventura’ received the lower clutch sizes and mean number of emerged adults. Mean developmental time (egg to adult) differed significantly among tested cultivars and were highest in ‘Camellia’ than in the other tested cultivars.  Only firmness and pH were correlated with SWD infestation as females tend to oviposit more eggs in softer fruits than in firmer fruits. Results also showed that a higher pH increased the emergence of adults and shortened the egg to adult developmental time.Research highlights: Our results showed significant differences in the susceptibility of SHB to SWD. This information may help design IPM programs and in making recommendations for blueberry crops as planting of low-chill cultivars expands

Sistema Automatizado de Lectura Visual de Datos de Medida

Adquisición de imágenes de un instrumento de medida con una webcam. Procesado y segmentación de esas imágenes para realizar una clasificación y obtener el dato medido.Image acquisition of a measuring instrument with a webcam. Processing and segmentation of these images for classifying and obtaining the measured data.Esteban Avivar, ÁE. (2013). Sistema Automatizado de Lectura Visual de Datos de Medida. http://hdl.handle.net/10251/33592.Archivo delegad

Inhibition of eIF2α dephosphorylation inhibits ErbB2-induced deregulation of mammary acinar morphogenesis

<p>Abstract</p> <p>Background</p> <p>The ErbB2/Her2/Neu receptor tyrosine kinase is amplified in ~30% of human breast cancers. Phosphorylation of the translation initiation factor, eIF2α inhibits global protein synthesis and activates a stress signaling and growth suppressive program. We have shown that forced phosphorylation of eIF2α can suppress head and neck, colorectal carcinoma and multiple myeloma tumor growth and/or survival. Here we explore whether ErbB2 modulates eIF2α phosphorylation and whether forced phosphorylation of the latter can antagonize ErbB2 deregulation of mammary acinar morphogenesis.</p> <p>Results</p> <p>We tested whether ErbB2 signaling influenced eIF2α signaling and whether enhanced phosphorylation of the latter affected ErbB2-deregulated mammary acinar development. We obtained stable MCF10A cells overexpressing wild-type (Wt) Neu/ErbB2 or a constitutively active (CA) variant via retroviral delivery or mammary tumor cells from MMTV-Neu tumors. Western blotting, RT-PCR and confocal microscopy were used to analyze the effects of ErbB2 activation on eIF2α signaling and the effect of the GADD34-PP1C inhibitor salubrinal. Wt- and MMTV-Neu cells formed aberrant acini structures resembling DCIS, while CA-ErbB2 overexpression induced invasive lesions. In these structures we found that CA-ErbB2 but not the Wt variant significantly down-regulated the pro-apoptotic gene CHOP. This occurred without apparent modulation of basal phosphorylation of PERK and eIF2α or induction of its downstream target ATF4. However, inhibition of eIF2α dephosphorylation with salubrinal was sufficient to inhibit Wt- and CA-ErbB2- as well as MMTV-Neu-induced deregulation of acinar growth. This was linked to enhanced CHOP expression, inhibition of proliferation, induction of apoptosis and luminal clearing in Wt-ErbB2 and to inhibition of cyclin D1 levels and subsequent proliferation in CA-ErbB2 cells.</p> <p>Conclusion</p> <p>Depending on the strength of ErbB2 signaling there is a differential regulation of CHOP and eIF2α phosphorylation. ErbB2 uncouples in basal conditions eIF2α phosphorylation from CHOP induction. However, this signal was restored by salubrinal treatment in Wt-ErbB2 expressing MCF10A cells as these DCIS-like structures underwent luminal clearing. In CA-ErbB2 structures apoptosis is not induced by salubrinal and instead a state of quiescence with reduced proliferation was achieved. Treatments that stabilize P-eIF2α levels may be effective in treating ErbB2 positive cancers without severely disrupting normal tissue function and structure.</p

Psychosocial Impairment as a Possible Cause of Sexual Dysfunction among Young Men with Mild Androgenetic Alopecia: A Cross-sectional Crowdsourcing Web-based Study

Finasteride 1 mg, one of the main treatments for male androgenetic alopecia (MAGA), may produce sexual dysfunction, but young men with MAGA could experience high psychosocial impairment because of changes in body image. Dissatisfaction with body image has been linked to an increase in problems with sexual function. To date no study has considered the possible effect of psychological impairment on sexual function of men with MAGA. Aim of our study was to explore the effect of psychosocial impairment produced by hair loss on the sexual function of men with MAGA. Patients and methods: Cross-sectional design. In total, 190 men with MAGA ranging 18-40 years old were recruited from an Internet online community. Participants completed an online survey comprising the SKINDEX-29 and the Massachusetts General Hospital Sex Functioning Questionnaire. Individuals with MAGA and moderate to severe psychosocial impairment had a higher risk of sexual dysfunction – adjusted odds ratio 2.1 (1.2-4.0; P=0.02) – compared with subjects with mild to absent impairment. Sexual desire and sexual arousal were the most affected elements of sexual response, but an increase in erectile dysfunction and reduced global satisfaction were also reported. We present the first study exploring the influence of MAGA psychosocial impairment on sexual function. Men between 18 and 40 years of age with moderate to severe MAGA psychosocial morbidity were found to have an increased risk of sexual dysfunction. Assessment of psychological morbidity and sexual function could be critical in patients with MAGA, particularly when considering treatment with finasteride.  </p

Dormancy Signatures and Metastasis in Estrogen Receptor Positive and Negative Breast Cancer

Breast cancers can recur after removal of the primary tumor and treatment to eliminate remaining tumor cells. Recurrence may occur after long periods of time during which there are no clinical symptoms. Tumor cell dormancy may explain these prolonged periods of asymptomatic residual disease and treatment resistance. We generated a dormancy gene signature from published experimental models and applied it to both breast cancer cell line expression data as well as four published clinical studies of primary breast cancers. We found that estrogen receptor (ER) positive breast cell lines and primary tumors have significantly higher dormancy signature scores (P<0.0000001) than ER- cell lines and tumors. In addition, a stratified analysis combining all ER+ tumors in four studies indicated 2.1 times higher hazard of recurrence among patients whose tumors had low dormancy scores (LDS) compared to those whose tumors had high dormancy scores (HDS) (p<0.000005). The trend was shown in all four individual studies. Suppression of two dormancy genes, BHLHE41 and NR2F1, resulted in increased in vivo growth of ER positive MCF7 cells. The patient data analysis suggests that disseminated ER positive tumor cells carrying a dormancy signature are more likely to undergo prolonged dormancy before resuming metastatic growth. Furthermore, genes identified with this approach might provide insight into the mechanisms of dormancy onset and maintenance as well as dormancy models using human breast cancer cell lines

A human tRNA methyltransferase 9-like protein prevents tumour growth by regulating LIN9 and HIF1-α

Emerging evidence points to aberrant regulation of translation as a driver of cell transformation in cancer. Given the direct control of translation by tRNA modifications, tRNA modifying enzymes may function as regulators of cancer progression. Here, we show that a tRNA methyltransferase 9‐like (hTRM9L/KIAA1456) mRNA is down‐regulated in breast, bladder, colorectal, cervix and testicular carcinomas. In the aggressive SW620 and HCT116 colon carcinoma cell lines, hTRM9L is silenced and its re‐expression and methyltransferase activity dramatically suppressed tumour growth in vivo. This growth inhibition was linked to decreased proliferation, senescence‐like G0/G1‐arrest and up‐regulation of the RB interacting protein LIN9. Additionally, SW620 cells re‐expressing hTRM9L did not respond to hypoxia via HIF1‐α‐dependent induction of GLUT1. Importantly, hTRM9L‐negative tumours were highly sensitive to aminoglycoside antibiotics and this was associated with altered tRNA modification levels compared to antibiotic resistant hTRM9L‐expressing SW620 cells. Our study links hTRM9L and tRNA modifications to inhibition of tumour growth via LIN9 and HIF1‐α‐dependent mechanisms. It also suggests that aminoglycoside antibiotics may be useful to treat hTRM9L‐deficient tumours.National Institute of Environmental Health Sciences (R01 ES015037)National Institute of Environmental Health Sciences (R01 ES017010)National Institute of Environmental Health Sciences (R21 ES017146)National Institute of Environmental Health Sciences (P30 ES002109)National Cancer Institute (U.S.) (R01 CA109182)National Cancer Institute (U.S.) (U54 CA163131)National Science Foundation (U.S.) (NSF 0922830)NYSTARWestaway Research FundSingapore-MIT Alliance for Research and TechnologySamuel Waxman Cancer Research Foundation Tumour Dormancy ProgramNYSTE

Phosphorylation of AIB1 at Mitosis Is Regulated by CDK1/CYCLIN B

Although the AIB1 oncogene has an important role during the early phase of the cell cycle as a coactivator of E2F1, little is known about its function during mitosis.Mitotic cells isolated by nocodazole treatment as well as by shake-off revealed a post-translational modification occurring in AIB1 specifically during mitosis. This modification was sensitive to the treatment with phosphatase, suggesting its modification by phosphorylation. Using specific inhibitors and in vitro kinase assays we demonstrate that AIB1 is phosphorylated on Ser728 and Ser867 by Cdk1/cyclin B at the onset of mitosis and remains phosphorylated until exit from M phase. Differences in the sensitivity to phosphatase inhibitors suggest that PP1 mediates dephosphorylation of AIB1 at the end of mitosis. The phosphorylation of AIB1 during mitosis was not associated with ubiquitylation or degradation, as confirmed by western blotting and flow cytometry analysis. In addition, luciferase reporter assays showed that this phosphorylation did not alter the transcriptional properties of AIB1. Importantly, fluorescence microscopy and sub-cellular fractionation showed that AIB1 phosphorylation correlated with the exclusion from the condensed chromatin, thus preventing access to the promoters of AIB1-dependent genes. Phospho-specific antibodies developed against Ser728 further demonstrated the presence of phosphorylated AIB1 only in mitotic cells where it was localized preferentially in the periphery of the cell.Collectively, our results describe a new mechanism for the regulation of AIB1 during mitosis, whereby phosphorylation of AIB1 by Cdk1 correlates with the subcellular redistribution of AIB1 from a chromatin-associated state in interphase to a more peripheral localization during mitosis. At the exit of mitosis, AIB1 is dephosphorylated, presumably by PP1. This exclusion from chromatin during mitosis may represent a mechanism for governing the transcriptional activity of AIB1

Analysis of Marker-Defined HNSCC Subpopulations Reveals a Dynamic Regulation of Tumor Initiating Properties

Head and neck squamous carcinoma (HNSCC) tumors carry dismal long-term prognosis and the role of tumor initiating cells (TICs) in this cancer is unclear. We investigated in HNSCC xenografts whether specific tumor subpopulations contributed to tumor growth. We used a CFSE-based label retentions assay, CD49f (α6-integrin) surface levels and aldehyde dehydrogenase (ALDH) activity to profile HNSCC subpopulations. The tumorigenic potential of marker-positive and -negative subpopulations was tested in nude (Balb/c nu/nu) and NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice and chicken embryo chorioallantoic membrane (CAM) assays. Here we identified in HEp3, SQ20b and FaDu HNSCC xenografts a subpopulation of G0/G1-arrested slow-cycling CD49fhigh/ALDH1A1high/H3K4/K27me3low subpopulation (CD49f+) of tumor cells. A strikingly similar CD49fhigh/H3K27me3low subpopulation is also present in primary human HNSCC tumors and metastases. While only sorted CD49fhigh/ALDHhigh, label retaining cells (LRC) proliferated immediately in vivo, with time the CD49flow/ALDHlow, non-LRC (NLRC) tumor cell subpopulations were also able to regain tumorigenic capacity; this was linked to restoration of CD49fhigh/ALDHhigh, label retaining cells. In addition, CD49f is required for HEp3 cell tumorigenicity and to maintain low levels of H3K4/K27me3. CD49f+ cells also displayed reduced expression of the histone-lysine N-methyltransferase EZH2 and ERK1/2phosphorylation. This suggests that although transiently quiescent, their unique chromatin structure is poised for rapid transcriptional activation. CD49f− cells can “reprogram” and also achieve this state eventually. We propose that in HNSCC tumors, epigenetic mechanisms likely driven by CD49f signaling dynamically regulate HNSCC xenograft phenotypic heterogeneity. This allows multiple tumor cell subpopulations to drive tumor growth suggesting that their dynamic nature renders them a “moving target” and their eradication might require more persistent strategies