Effect of climate change on the water quality of Mediterranean rivers and alternatives to improve its status.

Surface water (SW) quality is particularly vulnerable to increased concentrations of nutrients, and this issue may be exacerbated by climate change. Knowledge of the effects of temperature and rainfall on SW quality is required to take the necessary measures to achieve good SW status in the future. To address this, the aims of this study were threefold: (1) to assess how a changing climate may alter the nitrate, ammonium, phosphorus and biological oxygen demand status (BOD5) of SW; (2) assess the relationship between water quality and flow; and (3) simulate diffuse and point source pollution reduction scenarios in the Júcar River Basin District in the Mediterranean region. A regionalised long-term climate scenario was used following one Representative Concentration Pathway (RCP8.5) with the data incorporated into the coupling of hydrological and water quality models. According to these climate change scenarios, SW with poor nitrate, ammonium, phosphorus and BOD5 status are expected to increase in the future by factors of 1.3, 1.9, 4 and 4, respectively. Furthermore, median ammonium and phosphorus concentration may be doubled in months with low flows. Additional measures are required to maintain current status in the water bodies, and it is necessary to reduce at least 25% of diffuse nitrate pollution, and 50% of point loads of ammonium, phosphorus, and BOD5.

Serotypes and antimicrobial resistance of Streptococcus pneumoniae in children under 5 years of age pre- and post-pneumococcal conjugate vaccine introduction in Paraguay.

Introduction. Streptococcus pneumoniae remains a major cause of mortality and morbidity worldwide in children <5 years of age, even with advances in vaccination programmes.Hypothesis/Gap Statement. Reviewing and reporting trends in the distribution of pneumococcal serotypes and antimicrobial resistance in Paraguay will be useful for decision-making in public health.Aim. This study analysed the serotype distribution and antimicrobial resistance of S. pneumoniae and the characteristics of pneumococcal disease in children <5 years old before and after the introduction of pneumococcal conjugate vaccines (PCVs).Methodology. A total of 885 isolates and 278 S. pneumoniae PCR-positive clinical specimens were referred to the Central Laboratory of Public Health (LCSP) within the meningitis and pneumonia laboratory based-surveillance network in the period 2006-2020. Conventional and molecular microbiological techniques were used for confirmation and characterization.Results. We identified 563 cases of pneumococcal disease in the pre-vaccination period, 325 cases in the post-PCV10 period and 275 cases in the post-PCV13 period. The serotypes covered by PCV10 decreased from 78.6-6.5 %. However, additional serotypes covered by PCV13 increased from 6.6-57.5% and non-PCV13 serotypes increased from 14.8-36.0 % (P<0.001) in the post-PCV13 period. In cases of meningitis, the rate of resistance to penicillin decreased after the introduction of conjugate vaccines. No resistance to ceftriaxone was found in any period. In cases without meningitis, the rate of resistance to penicillin and ceftriaxone decreased slightly. However, the rate of resistance to erythromycin and tetracycline increased and that to trimethoprim-sulfamethoxazole (SXT) decreased in the post-PCV13 period compared to the pre-PCV period. The multidrug resistance rate was 8.5 %.Conclusion. A change in the circulating serotypes and antimicrobial resistance to certain antibiotics was observed. Non-vaccine serotype circulation and multidrug resistance may compromise the success of the conjugate vaccines.

Enhancement of the CO2 Sensing/Capture through High Cationic Charge in M-ZrO2 (Li+, Mg2+, or Co3+): Experimental and Theoretical Studies.

The capture and storage of CO2 are of growing interest in atmospheric science since greenhouse gas emission has to be reduced considerably in the near future. The present paper deals with the doping of cations on ZrO2, i.e., M-ZrO2 (M = Li+, Mg2+, or Co3+), defecting the crystalline planes for the adsorption of carbon dioxide. The samples were prepared by the sol-gel method and characterized completely by different analytical methods. The deposition of metal ions on ZrO2 (whose crystalline phases: monoclinic and tetragonal are transformed into a single-phase such as tetragonal for LiZrO2 and cubic for MgZrO2 or CoZrO2) shows a complete disappearance of the XRD monoclinic signal, and it is consistent with HRTEM lattice fringes: 2.957 nm for ZrO2 (101, tetragonal/monoclinic), 3.018 nm for tetragonal LiZrO2, 2.940 nm for cubic MgZrO2, and 1.526 nm for cubic CoZrO2. The samples are thermally stable, resulting an average size of ∼5.0-15 nm. The surface of LiZrO2 creates the oxygen deficiency, while for Mg2+ (0.089 nm), since the size of the atom is relatively greater than that of Zr4+ (0.084 nm), the replacement of Zr4+ by Mg2+ in sublattice is difficult; thus, a decrease of the lattice constant was noticed. Since the high band gap energy (ΔE > 5.0 eV) is suitable for CO2 adsorption, the samples were employed for the selective detection/capture of CO2 by using electrochemical impedance spectroscopy (EIS) and direct current resistance (DCR), showing that CoZrO2 is capable of CO2 capture about 75%. If M+ ions are deposited within the ZrO2 matrix, then the charge imbalance allows CO2 to interact with the oxygen species to form CO32- which produces a high resistance (21.04 × 106 (Ω, Ohm)). The adsorption of CO2 with the samples was also theoretically studied showing that the interaction of CO2 with MgZrO2 and CoZrO2 is more feasible than with LiZrO2, subscribing to the experimental data. The temperature effect (273 to 573 K) for the interaction of CO2 with CoZrO2 was also studied by the docking method and observed the cubic structure is more stable at high temperatures as compared to the monoclinic geometry. Thus, CO2 would preferably interact with ZrO2c (ERS = -19.29 kJ/mol) than for ZrO2m (22.4 J/mmol (ZrO2c = cubic; ZrO2m = monoclinic).

Serogrupos y resistencia antimicrobiana de Neisseria meningitidis causantes de enfermedad invasiva en Paraguay

RESUMEN La enfermedad meningocócica representa un problema de salud pública y una de las principales causas de morbilidad y mortalidad en todo el mundo. Los serogrupos que causan la mayor carga de enfermedad a nivel global son A, B, C, W e Y. El objetivo del estudio fue describir los serogrupos y la resistencia antimicrobiana de Neisseria meningitidis aisladas de enfermedad invasiva en Paraguay durante el periodo 2010-2020. Se estudiaron todas las muestras de líquido cefalorraquídeo y sangre con aislamientos o detección de ADN por PCR de N. meningitidis de pacientes de diversas edades, que fueron remitidas al Laboratorio Central de Salud Pública durante el periodo 2010-2020, dentro del marco de la vigilancia epidemiológica de meningitis (n=163) en Paraguay. La mayor frecuencia de hallazgos de N. meningitidis se observó en el grupo de edad de < 1 año. El 25,7% de los casos correspondió al serogrupo B, el 52,1% al serogrupo C, 18,4% al serogrupo W y 3,7% al serogrupo Y. En el 2018, se evidenció la mayor cantidad de casos por serogrupo C (n=27). La menor frecuencia de sensibilidad disminuida a penicilina G fue en el 2010 (12,5%) y la mayor en el 2014 (100,0%). Se registró un aumento de casos por serogrupo C a partir del 2017, posicionándose como serogrupo prevalente hasta el 2020, y además, un aumento de la sensibilidad disminuida a la penicilina. La vigilancia es de importancia en el control de la enfermedad meningocócica para detección de brotes, estimación de la carga de enfermedad, análisis de resistencia antimicrobiana, distribución de serogrupos y evaluaciones de estrategias de control.

ABSTRACT Meningococcal disease represents a public health problem and one of the main causes of morbidity and mortality worldwide. The serogroups that cause the highest burden of disease globally are A, B, C, W, and Y. The aim of the study was to describe serogroups and antimicrobial resistance of Neisseria meningitidis isolated from invasive disease in Paraguay during the 2010-2020 period. Samples of cerebrospinal fluid and blood with isolates or detection of DNA by PCR of N. meningitidis from patients of different ages referred to the Central Public Health Laboratory during the period 2010-2020 within the framework of the epidemiological surveillance of meningitis (n = 163) in Paraguay were studied. The highest frequency of N. meningitidis findings was observed in the <1 year age group, 25.7% of the cases corresponded to serogroup B, 52.1% to serogroup C, 18.4% to serogroup W and 3.7% to serogroup Y. In 2018, the highest number of cases by serogroup C (n = 27) was found. The lowest frequency of decreased sensitivity to penicillin G was in 2010 (12.5%) and the highest in 2014 (100.0%). There was an increase in cases due to serogroup C in 2017, positioning itself as the prevalent serogroup until 2020, in addition, there was an increase in decreased sensitivity to penicillin. Surveillance is important in the control of meningococcal disease for outbreak detection, estimation of the burden of disease, analysis of antimicrobial resistance, serogroup distribution, and evaluations of control strategies.

Identifying resilient industries in Mexico's automotive cluster: Policy lessons from the great recession to surmount the crisis caused by COVID 19.

This study investigates regional employment and production resilience to adverse crisis shocks. We extend the concept of resilience beyond the notion of resistance and fast recovery to include the potential of positive spillovers to related industries in neighboring regions. The industrial analysis of the automotive cluster in Mexico relies on a spatial shift-share approach to characterize the resilience ability of regional industries. The study employs the Sub-Prime crisis as a benchmark to identify the type of employment and production recovery after an adverse shock of similar magnitude. Four industries within the automotive cluster present various degrees of spatial dependence-those industries with the most significant spillover effects on neighboring regions. The type of resilience shown by these automotive industries in the regions during the Sub Prime Crisis serves as a benchmark to base policy government and corporate decisions after the economic crisis caused by the outbreak of SARS-CoV-2.

Transposon mutagenesis identifies cooperating genetic drivers during keratinocyte transformation and cutaneous squamous cell carcinoma progression.

The systematic identification of genetic events driving cellular transformation and tumor progression in the absence of a highly recurrent oncogenic driver mutation is a challenge in cutaneous oncology. In cutaneous squamous cell carcinoma (cuSCC), the high UV-induced mutational burden poses a hurdle to achieve a complete molecular landscape of this disease. Here, we utilized the Sleeping Beauty transposon mutagenesis system to statistically define drivers of keratinocyte transformation and cuSCC progression in vivo in the absence of UV-IR, and identified both known tumor suppressor genes and novel oncogenic drivers of cuSCC. Functional analysis confirms an oncogenic role for the ZMIZ genes, and tumor suppressive roles for KMT2C, CREBBP and NCOA2, in the initiation or progression of human cuSCC. Taken together, our in vivo screen demonstrates an extremely heterogeneous genetic landscape of cuSCC initiation and progression, which can be harnessed to better understand skin oncogenic etiology and prioritize therapeutic candidates.

Estudio epidemiológico de Haemophilus influenzae causante de enfermedad invasiva y no invasiva en Paraguay (1999-2017) / Epidemiologic study of Haemophilus influenzae causing invasive and non-invasive disease in Paraguay (1999-2017)

Introducción: Haemophilus influenzae (Hi) causa infecciones leves e invasivas graves, especialmente en niños menores de 5 años. El serotipo b (Hib) era muy frecuente antes de la introducción de la vacuna, que en Paraguay se introdujo en 2004. Métodos: Se estudiaron por métodos microbiológicos convencionales y técnicas moleculares 523 aislados de H. influenzae obtenidos desde 1999 a 2017 y remitidos al Laboratorio Nacional de Referencia de Paraguay. Resultados: El serotipo más frecuente fue el no-tipable (HiNT), en un 51,8% (271/523), seguido de Hib (43%; 225/523), Hia e Hif (1,5%; 8/523, respectivamente), Hic (1%; 5/523), Hie (0,8%; 4/523) y Hid (0,4%; 2/523). El 48,4% de las infecciones invasivas estaban causadas por HiNT y el 46,1% por Hib. El 88,6% de las cepas procedían de casos de meningitis, el 70,8% de sepsis y el 50,9% de neumonías en niños menores de 5 años. El 84% (181/217) de las cepas correspondían a enfermedades invasivas causadas por Hib en menores de 5 años, registrándose la mayor proporción entre 2001-2003. Los biotipos más prevalentes fueron el biotipo I (29%), biotipo II (12%), biotipo III (24%) y biotipo IV (13%). El 13% (68/523) de los aislados fueron resistentes a la ampicilina. Conclusión: Tras de la introducción de la vacuna frente a Hib en Paraguay, los casos de enfermedad invasiva por Hib disminuyeron en niños menores de 5 años y hubo un aumento de HiNT en los mayores de 5 años. Es necesaria una vigilancia continua para monitorizar la eficacia de la vacuna y desarrollar intervenciones preventivas.(AU)

Introduction: Haemophilus influenzae is a cause of mild and severe invasive infections, especially among children under 5 years old. Serotype b (Hib) was very frequent before the introduction of the vaccine, which was introduced in Paraguay in 2004. Methods: A total of 523 isolates of H. influenzae obtained from 1999 to 2017 and referred to the National Reference Laboratory in Paraguay were studied by conventional microbiological methods and molecular techniques. Results: The most frequent serotype was non-typeable (HiNT) (51.8%; 271/523), followed by Hib (43%; 225/523), Hia and Hif (1.5%; 8/523, respectively), Hic (1%; 5/523), Hie (0.8%; 4/523), and Hid (0.4%; 2/523). A total of 48.4% invasive infections were caused by HiNT, and 46.1% by Hib; 88.6% of isolates corresponded to meningitis, 70.8% to sepsis and 50.9% to pneumonia in children under 5 years. A total of 84% (181/217) of isolates corresponded to invasive infections caused by Hib in children under 5 years, with the highest proportion observed between 2001 and 2003. The most prevalent biotypes were biotype I (29%), biotype II (12%), biotype III (24%), and biotype IV (13%). Among the total of isolates, 13% (68/523) of isolates were resistant to ampicillin. Conclusion: After the introduction of the Hib vaccine in Paraguay, the number of invasive Hib cases decreased in children under 5 years old, although we observed an increase of HiNT in children over 5 years. Continuous surveillance is necessary in order to monitor the effectiveness of the vaccine and for the development of preventive interventions.(AU)

Epidemiologic study of Haemophilus influenzae causing invasive and non-invasive disease in Paraguay (1999-2017). / Estudio epidemiológico de Haemophilus influenzae causante de enfermedad invasiva y no invasiva en Paraguay (1999-2017).

INTRODUCTION: Haemophilus influenzae is a cause of mild and severe invasive infections, especially among children under 5 years old. Serotype b (Hib) was very frequent before the introduction of the vaccine, which was introduced in Paraguay in 2004. METHODS: A total of 523 isolates of H. influenzae obtained from 1999 to 2017 and referred to the National Reference Laboratory in Paraguay were studied by conventional microbiological methods and molecular techniques. RESULTS: The most frequent serotype was non-typeable (HiNT) (51.8%; 271/523), followed by Hib (43%; 225/523), Hia and Hif (1.5%; 8/523, respectively), Hic (1%; 5/523), Hie (0.8%; 4/523), and Hid (0.4%; 2/523). A total of 48.4% invasive infections were caused by HiNT, and 46.1% by Hib; 88.6% of isolates corresponded to meningitis, 70.8% to sepsis and 50.9% to pneumonia in children under 5 years. A total of 84% (181/217) of isolates corresponded to invasive infections caused by Hib in children under 5 years, with the highest proportion observed between 2001 and 2003. The most prevalent biotypes were biotype I (29%), biotype II (12%), biotype III (24%), and biotype IV (13%). Among the total of isolates, 13% (68/523) of isolates were resistant to ampicillin. CONCLUSION: After the introduction of the Hib vaccine in Paraguay, the number of invasive Hib cases decreased in children under 5 years old, although we observed an increase of HiNT in children over 5 years. Continuous surveillance is necessary in order to monitor the effectiveness of the vaccine and for the development of preventive interventions.

Frecuencia de Streptococcus pneumoniae aislados de enfermedad invasiva en Paraguay, serotipos y perfil de sensibilidad (2010-2018) / Frequency of Streptococcus pneumoniae isolated from invasive disease in Paraguay, serotypes and sensitivity profile (2010-2018)

Streptococcus pneumoniae sigue siendo una de las causas más importantes de morbilidad y mortalidad en niños y adultos alrededor del mundo. El objetivo del estudio fue describir la frecuencia de aislamiento de S. pneumoniae en enfermedad invasiva, distribución de serotipos y sensibilidad antimicrobiana en Paraguay (2010-2018). Se estudiaron 793 cepas de S. pneumoniae aisladas de pacientes de todas las edades con enfermedad invasiva en Paraguay, provenientes de los diferentes centros centinelas y colaboradores en el marco de la vigilancia de meningitis y neumonías, durante el periodo 2010-2018. La frecuencia general según diagnóstico resultó 74.9% de neumonías (n=594), 18.4% de meningitis (n=146) y 6.7% de sepsis (n=53). El serotipo 14 fue más frecuente con 174 aislamientos (22.0%), seguido del serotipo 19A con 84 aislamientos (10.6%), el serotipo 3 con 66 aislamientos (8.3%) y el 6A con 37 aislamientos (4.7%). En meningitis se registró una frecuencia general de resistencia a penicilina del 32,2% y de ceftriaxona del 1,4%. En los casos de no meningitis la resistencia a penicilina fue del 0,8% y ceftriaxona del 0,3%. Los resultados de serotipos y sensibilidad antimicrobiana proporcionarán información necesaria para la implementación de estrategias de prevención y tratamiento de la enfermedad neumocócica en nuestro país, por lo que es necesaria una vigilancia continua para evaluar la carga de enfermedad, los serotipos circulantes y el aumento de la resistencia a los antibióticos

Streptococcus pneumoniae remains one of the most important causes of morbidity and mortality in children and adults worldwide. The objective of the study was to describe the frequency of isolation of S. pneumoniae in invasive disease, serotype distribution and antimicrobial susceptibility in Paraguay (2010-2018). We studied 793 strains of S. pneumoniae isolated from patients of all ages with invasive disease in Paraguay, from different sentinel centers and collaborators in the framework of meningitis and pneumonia surveillance during the period 2010-2018. The general frequency according to diagnosis was 74.9% of pneumonia (n = 594), 18.4% of meningitis (n = 146) and 6.7% of sepsis (n = 53). Serotype 14 was more frequent with 174 isolates (22.0%), followed by serotype 19A with 84 isolates (10.6%), serotype 3 with 66 isolates (8.3%) and 6A with 37 isolates (4.70%). In meningitis, there was a general frequency of penicillin resistance of 32.2% and ceftriaxone of 1.4%. In cases of non-meningitis, penicillin resistance was 0.8% and ceftriaxone 0.3%. The results of serotypes and antimicrobial sensitivity will provide necessary information for the implementation of prevention strategies and treatment of pneumococcal disease in our country, therefore it is necessary to continue monitoring in order to assess the burden of the disease, circulating serotypes and increased antibiotic resistance

Análisis de tareas para las funciones espaciales en niños escolares con diagnóstico de TDAH / Analysis of Tasks Used for Assessment of Spatial Function in Schoolchildren with ADHD / Análise de tarefas para as funções espaciais em crianças escolares com diagnóstico de TDAH

Resumen Objetivo. Identificar y precisar las dificultades en funciones espaciales que presentan los niños con TDAH en edad escolar. Método. Se evaluaron 24 niños, entre los 6 y los 12 años de edad, con diagnóstico de TDAH. Todos los niños fueron evaluados con tareas gráficas, tareas verbales y tareas constructivas, incluidas en las pruebas Evaluación Neuropsicológica Infantil, Evaluación Neuropsicológica de la Integración Espacial y Evaluación Neuropsicológica del Éxito Escolar. Resultados. El análisis clínico realizado al desempeño de los niños mostró dificultades no solo en el manejo de la proporción, la integración y al dibujar ángulos, sino en el uso adecuado del espacio e inversión. Las dificultades espaciales estuvieron acompañadas de otras de índole regulatorio, tono de activación, motoras y cinestésicas. Los participantes con mayor número de errores fueron aquellos de menor edad. Conclusión. Los niños con diagnóstico de TDAH pueden presentar dificultades en el desarrollo de las funciones espaciales, por lo cual se hace necesario que la evaluación neuropsicológica incluya un análisis profundo de esta dimensión cognitiva, que permita indagar la naturaleza de estas dificultades y su posible relación con el desarrollo de otras funciones.

Abstract Objective. Identify and detail the difficulties in children with attention deficit disorder in spatial function tasks. Method. 24 school children between 6 and 12 years old with diagnosis of attention deficit disorder were included in the study. Neuropsychological tasks used in the study were divided in graphic tasks, verbal tasks and a constructive task, all of them included in the Neuropsychological Evaluation for Children, Neuropsychological Evaluation of the Spatial Integration and Neuropsychological Evaluation of School Success assessments. Results. Analysis of the results has pointed out specific errors in graphic tasks such as disproportion, disintegration, difficulties with angles, problems with usage of space and inversions. Younger participants with complex neuropsychological syndromes committed more mistakes. The spatial difficulties were accompanied by regulatory, activation, motor and kinesthetic difficulties. The participants with higher frequencies of error were those of lower age. Conclusion. The results show that children with attention deficit disorder diagnosis may have difficulties in the development of spatial function; therefore, it is necessary to include a more profound neuropsychological analysis of the domain that allows the investigation of the nature of the difficulties and its plausible relation to the development of other functions.

Resumo Escopo. Identificar e reconhecer as dificuldades em funções espaciais que apresentam as crianças com TDAH em idade escolar. Metodologia. Foram avaliadas 20 crianças masculinas e 4 femininas, com idades compreendidas entre os 6 e 12 anos de idade, com diagnóstico de TDAH. Todas as crianças foram avaliadas com tarefas gráficas, tarefas verbais e tarefas construtivas, incluídas nas provas Avaliação Neuropsicológica da Integração espacial e Avaliação Neuropsicológica do Sucesso Escolar. Resultados. A análise clínica realizada aos desempenhos das crianças, mostraram dificuldades de desproporção, desintegração, dificuldade para desenhar ângulos, uso inadequado do espaço e inversão. As dificuldades espaciais estiveram acompanhadas de outras de natureza regulatória, tono de ativação, motoras e cinestésicas. Os participantes com maior número de erros foram aqueles de menor idade. Conclusão. As crianças com diagnóstico de déficit de atenção podem apresentar dificuldades no desenvolvimento das funções espaciais, pelo qual é preciso que a avaliação neuropsicológica possa incluir uma análise profunda de esta dimensão cognitiva, que permita indagar a natureza de estas dificuldades e sua possível relação com o desenvolvimento de outras funções.

[Genotypes of Neisseria meningitidis isolates in patients with meningococcal meningitis in Paraguay, 1996-2015Genótipos de Neisseria meningitidis isolados de pacientes com doença meningocócica no Paraguai, 1996-2015]. / Genotipos de Neisseria meningitidis aislados de pacientes con enfermedad meningocócica en Paraguay, 1996-2015.

OBJECTIVE: Describe the phenotypical and genotypical characteristics of Neisseria meningitidis isolates from cases of meningococcal disease in Paraguay between 1996 and 2015. METHODS: Conventional microbiological methods and molecular techniques were used to study 114 isolates of N. meningitidis and 12 clinical samples without isolation (confirmed by polymerase chain reaction), provided by various sentinel centers and collaborating centers in Paraguay. RESULTS: The most affected age group was children under 1 year (19.0%), followed by 1-5-year-olds (17.5%). The highest percentage of strains was isolated in meningitis cases (81.7%) and serogroup B was found in 60.3% of cases. The most frequent phenotypes were B:4:p1.14 (16.0%), B:15:p1.5, C:nt:nst, and W:nt:p1.2 (10.5%), respectively. The prevalent clonal complexes were: ST-11/ET37 complex, 29.6% (8/27), predominantly serogroup W (6/8); ST-35 complex, 18.5% (5/27), in serogroup B (4/4); and ST-32/ET5 complex, 14.8% (4/16), in serogroup B (5/5). CONCLUSIONS: Meningococcal meningitis is relatively uncommon in Paraguay. Distribution analysis showed that serogroup B is the most common and that the number of cases of meningococcal disease caused by serogroups C and W increased in the last two years. The identified clonal complexes were correlated with those found in the Southern Cone region. Due to the high virulence of N. meningitidis, its surveillance should be a strategic priority of national and regional public health systems to prevent epidemic outbreaks and support public health decision-making.

OBJETIVO: Descrever as características fenotípicas e genotípicas de cepas de Neisseria meningitidis isoladas de casos de doença meningocócica no Paraguai entre 1996 e 2015. MÉTODOS: Foram estudados por métodos microbiológicos convencionais e técnicas moleculares 114 isolados de N. meningitidis e 12 amostras clínicas sem isolamento confirmadas por reação em cadeia da polimerase (PCR) enviados por diferentes centros-sentinela e centros colaboradores do Paraguai. RESULTADOS: A faixa etária mais afetada foi a de crianças menores de 1 ano (19,0%) e crianças de 1 a 5 anos (17,5%). Uma maior porcentagem de cepas foi isolada de casos de meningite (81,7%) e o sorogrupo B foi identificado em 60,3% dos casos. Os fenótipos mais comuns foram B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST e W:NT:P1.2 (10,5%), respectivamente. Os complexos clonais mais prevalentes foram o complexo ST-11/ET37 (29,6%, 8/27) com predomínio no sorogrupo W (6/8), complexo ST-35 (18,5%, 5/27) no sorogrupo B (4/4) e complexo ST-32/ET5 (14,8%, 4/16) no sorogrupo B (5/5). CONCLUSÕES: A doença meningocócica é relativamente pouco comum no Paraguai. A análise da distribuição dos sorogrupos demonstrou que o sorogrupo B é o mais prevalente e, nos últimos dois anos, ouve um aumento nos casos de doença meningocócica pelos sorogrupos C e W. Os complexos clonais encontrados se correlacionam com os achados na região do Cone Sul. Devido à alta virulência da N. meningitidis, a vigilância deste agente deve ser uma prioridade estratégica dos sistemas de saúde pública nacionais e regionais para prevenir surtos epidêmicos e subsidiar a tomada de decisão em saúde pública.

Mean platelet volume as a prognostic factor for venous thromboembolic disease.

BACKGROUND: Mean platelet volume (MPV) is a risk factor for cardiovascular and inflammatory diseases. AIM: To evaluate the association between high MPV and 90-day mortality after an episode of venous thromboembolism (VTE). MATERIAL AND METHODS: Retrospective cohort of 594 patients with a median age of 73 years (58% women) with a first episode VTE, included in an institutional Thromboembolic Disease registry between 2014 and 2015. MPV values were obtained from the automated blood cell count measured at the moment of VTE diagnosis. Volumes ≥ 11 fL were classified as high. All patients were followed for 90 days to assess survival. RESULTS: The main comorbidities were cancer in 221 patients (37%), sepsis in 172 (29%) and coronary artery disease in 107 (18%). Median MPV was 8 fl (8-9), brain natriuretic peptide 2,000 pg/ml (1,025-3,900) and troponin 40 pg/ml (19.5-75). Overall mortality was 20% (121/594) during the 90 days of follow-up. Thirty three deaths occurred within 7 days and 43 within the first month. The loss of patients from follow-up was 5% (28/594) at 90 days. Mortality among patients with high MP was 36% (23/63). The crude mortality hazard ratio (HR) for high MPV was 2.2 (95% confidence intervals (CI) 1.4-3.5). When adjusted for sepsis, oncological disease, heart disease, kidney failure and surgery, the mortality HR of high MPV was 2.4 (CI95% 1.5-3.9) in the VTE group, 2.3 (CI95% 1.5-4.4) in the deep venous thrombosis group, and 2.9 (CI95% 1.6 -5.6) in the pulmonary embolism group. CONCLUSIONS: High MPV is an independent risk factor for mortality following an episode of VTE.

Mean platelet volume as a prognostic factor for venous thromboembolic disease / Volumen plaquetario medio como factor pronóstico de la enfermedad tromboembólica venosa

ABSTRACT Background: Mean platelet volume (MPV) is a risk factor for cardiovascular and inflammatory diseases. Aim: To evaluate the association between high MPV and 90-day mortality after an episode of venous thromboembolism (VTE). Material and Methods: Retrospective cohort of 594 patients with a median age of 73 years (58% women) with a first episode VTE, included in an institutional Thromboembolic Disease registry between 2014 and 2015. MPV values were obtained from the automated blood cell count measured at the moment of VTE diagnosis. Volumes ≥ 11 fL were classified as high. All patients were followed for 90 days to assess survival. Results: The main comorbidities were cancer in 221 patients (37%), sepsis in 172 (29%) and coronary artery disease in 107 (18%). Median MPV was 8 fl (8-9), brain natriuretic peptide 2,000 pg/ml (1,025-3,900) and troponin 40 pg/ml (19.5-75). Overall mortality was 20% (121/594) during the 90 days of follow-up. Thirty three deaths occurred within 7 days and 43 within the first month. The loss of patients from follow-up was 5% (28/594) at 90 days. Mortality among patients with high MP was 36% (23/63). The crude mortality hazard ratio (HR) for high MPV was 2.2 (95% confidence intervals (CI) 1.4-3.5). When adjusted for sepsis, oncological disease, heart disease, kidney failure and surgery, the mortality HR of high MPV was 2.4 (CI95% 1.5-3.9) in the VTE group, 2.3 (CI95% 1.5-4.4) in the deep venous thrombosis group, and 2.9 (CI95% 1.6 −5.6) in the pulmonary embolism group. Conclusions: High MPV is an independent risk factor for mortality following an episode of VTE.

Antecedentes: El volumen plaquetario medio (VPM) es un factor de riesgo de complicaciones cardiovasculares y enfermedades inflamatorias. Objetivo: Evaluar la asociación entre VPM alto y la mortalidad a los 90 días después de un episodio de tromboembolismo venoso (ETV). Material y Métodos: Cohorte retrospectiva de 594 pacientes adultos con una edad media de 73 años (58% mujeres) con un primer episodio de ETV incluidos en un registro de enfermedad tromboembólica institucional entre 2014 y 2015. Se obtuvieron valores de VPM desde el hemograma tomado en el momento del diagnóstico de ETV y un volumen ≥ 11 fL fue clasificado como alto. Todos los pacientes fueron seguidos durante 90 días para determinar sobrevida. Resultados: Las comorbilidades fueron cáncer en 221 pacientes (37%), sepsis en 172 (29%) y enfermedad coronaria en 107 (18%). La mediana de VPM fue 8 fl (89), el péptido natriurético cerebral fue de 2.000 pg/ml (1.025-3.900) y la troponina fue de 40 pg/ml (19,5-75). La mortalidad global a 90 días fue 20% (121/594). Treinta y tres muertes ocurrieron dentro de los 7 días y 43 en el primer mes. La pérdida de seguimiento de pacientes fue de 5% (28/594) a los 90 días. La mortalidad en el grupo con VPM alto fue 36% (23/63). La razón de riesgo (HR) cruda de la mortalidad para un VPM alto fue de 2,2 (intervalos de confianza (IC) de 95% 1,4-3,5). Cuando se ajustó por sepsis, enfermedad oncológica, enfermedad cardíaca, insuficiencia renal y cirugía, la HR de muerte para un VPM alto fue de 2,4 (IC95% 1,5-3,9) en el grupo de ETV; 2,3 (IC95% 1,5-4,4) en el grupo de trombosis venosa profunda; y 2,9 (CI95% 1,6 −5,6) en el grupo de embolia pulmonar. Conclusiones: Un VPM alto es un factor de riesgo independiente de mortalidad después de un episodio de ETV.

Genotipos de Neisseria meningitidis aislados de pacientes con enfermedad meningocócica en Paraguay, 1996–2015

[RESUMEN]. Objetivo. Describir las características fenotípicas y genotípicas de cepas de Neisseria meningitidis aisladas de enfermedad meningocócica en Paraguay entre 1996 y 2015. Métodos. Se estudiaron por métodos microbiológicos convencionales y técnicas moleculares 114 aislamientos de N. meningitidis y 12 muestras clínicas sin aislamiento confirmadas por reacción en cadena de la polimerasa (PCR) que fueron remitidas por los diferentes centros centinelas y centros colaboradores de Paraguay. Resultados. El grupo de edad más afectado fue el de menores de 1 año (19,0%), seguido por el de 1 a 5 años (17,5%). Un mayor porcentaje de las cepas se aisló de casos de meningitis (81,7%) y el serogrupo B se encontró en 60,3% de los casos. Los fenotipos más frecuentes fueron B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST y W:NT:P1.2 (10,5%), respectivamente. Los complejos clonales prevalentes fueron ST-11/ET37 complex 29,6% (8/27) con predominio del serogrupo W (6/8), ST-35 complex 18,5% (5/27) en el serogrupo B (4/4), y ST-32/ET5 complex 14,8% (4/16) en el serogrupo B (5/5). Conclusiones. En Paraguay la enfermedad meningocócica es relativamente infrecuente. Los análisis de distribución de serogrupo muestran que el más frecuente es el B y en los últimos dos años aumentaron los casos de enfermedad meningocócica por C y W. Los complejos clonales encontrados se correlacionan con los hallados en la región del Cono Sur. Debido al alto nivel de virulencia de N. meningitidis, su vigilancia debe constituir una prioridad estratégica de los sistemas de salud pública nacionales y regionales para prevenir brotes epidémicos y apoyar la toma de decisiones en salud pública.

[ABSTRACT]. Objective. Describe the phenotypical and genotypical characteristics of Neisseria meningitidis isolates from cases of meningococcal disease in Paraguay between 1996 and 2015. Methods. Conventional microbiological methods and molecular techniques were used to study 114 isolates of N. meningitidis and 12 clinical samples without isolation (confirmed by polymerase chain reaction), provided by various sentinel centers and collaborating centers in Paraguay. Results. The most affected age group was children under 1 year (19.0%), followed by 1-5-year-olds (17.5%). The highest percentage of strains was isolated in meningitis cases (81.7%) and serogroup B was found in 60.3% of cases. The most frequent phenotypes were B:4:p1.14 (16.0%), B:15:p1.5, C:nt:nst, and W:nt:p1.2 (10.5%), respectively. The prevalent clonal complexes were: ST-11/ET37 complex, 29.6% (8/27), predominantly serogroup W (6/8); ST-35 complex, 18.5% (5/27), in serogroup B (4/4); and ST-32/ET5 complex, 14.8% (4/16), in serogroup B (5/5). Conclusions. Meningococcal meningitis is relatively uncommon in Paraguay. Distribution analysis showed that serogroup B is the most common and that the number of cases of meningococcal disease caused by serogroups C and W increased in the last two years. The identified clonal complexes were correlated with those found in the Southern Cone region. Due to the high virulence of N. meningitidis, its surveillance should be a strategic priority of national and regional public health systems to prevent epidemic outbreaks and support public health decision-making.

[RESUMO]. Objetivo. Descrever as características fenotípicas e genotípicas de cepas de Neisseria meningitidis isoladas de casos de doença meningocócica no Paraguai entre 1996 e 2015. Métodos. Foram estudados por métodos microbiológicos convencionais e técnicas moleculares 114 isolados de N. meningitidis e 12 amostras clínicas sem isolamento confirmadas por reação em cadeia da polimerase (PCR) enviados por diferentes centrossentinela e centros colaboradores do Paraguai. Resultados. A faixa etária mais afetada foi a de crianças menores de 1 ano (19,0%) e crianças de 1 a 5 anos (17,5%). Uma maior porcentagem de cepas foi isolada de casos de meningite (81,7%) e o sorogrupo B foi identificado em 60,3% dos casos. Os fenótipos mais comuns foram B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST e W:NT:P1.2 (10,5%), respectivamente. Os complexos clonais mais prevalentes foram o complexo ST-11/ET37 (29,6%, 8/27) com predomínio no sorogrupo W (6/8), complexo ST-35 (18,5%, 5/27) no sorogrupo B (4/4) e complexo ST-32/ET5 (14,8%, 4/16) no sorogrupo B (5/5). Conclusões. A doença meningocócica é relativamente pouco comum no Paraguai. A análise da distribuição dos sorogrupos demonstrou que o sorogrupo B é o mais prevalente e, nos últimos dois anos, ouve um aumento nos casos de doença meningocócica pelos sorogrupos C e W. Os complexos clonais encontrados se correlacionam com os achados na região do Cone Sul. Devido à alta virulência da N. meningitidis, a vigilância deste agente deve ser uma prioridade estratégica dos sistemas de saúde pública nacionais e regionais para prevenir surtos epidêmicos e subsidiar a tomada de decisão em saúde pública.

Genotipos de Neisseria meningitidis aislados de pacientes con enfermedad meningocócica en Paraguay, 1996-2015 / Genotypes of Neisseria meningitidis isolates in patients with meningococcal meningitis in Paraguay, 1996-2015 / Genótipos de Neisseria meningitidis isolados de pacientes com doença meningocócica no Paraguai, 1996-2015

RESUMEN Objetivo Describir las características fenotípicas y genotípicas de cepas de Neisseria meningitidis aisladas de enfermedad meningocócica en Paraguay entre 1996 y 2015. Métodos Se estudiaron por métodos microbiológicos convencionales y técnicas moleculares 114 aislamientos de N. meningitidis y 12 muestras clínicas sin aislamiento confirmadas por reacción en cadena de la polimerasa (PCR) que fueron remitidas por los diferentes centros centinelas y centros colaboradores de Paraguay. Resultados El grupo de edad más afectado fue el de menores de 1 año (19,0%), seguido por el de 1 a 5 años (17,5%). Un mayor porcentaje de las cepas se aisló de casos de meningitis (81,7%) y el serogrupo B se encontró en 60,3% de los casos. Los fenotipos más frecuentes fueron B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST y W:NT:P1.2 (10,5%), respectivamente. Los complejos clonales prevalentes fueron ST-11/ET37 complex 29,6% (8/27) con predominio del serogrupo W (6/8), ST-35 complex 18,5% (5/27) en el serogrupo B (4/4), y ST-32/ET5 complex 14,8% (4/16) en el serogrupo B (5/5). Conclusiones En Paraguay la enfermedad meningocócica es relativamente infrecuente. Los análisis de distribución de serogrupo muestran que el más frecuente es el B y en los últimos dos años aumentaron los casos de enfermedad meningocócica por C y W. Los complejos clonales encontrados se correlacionan con los hallados en la región del Cono Sur. Debido al alto nivel de virulencia de N. meningitidis, su vigilancia debe constituir una prioridad estratégica de los sistemas de salud pública nacionales y regionales para prevenir brotes epidémicos y apoyar la toma de decisiones en salud pública.

ABSTRACT Objective Describe the phenotypical and genotypical characteristics of Neisseria meningitidis isolates from cases of meningococcal disease in Paraguay between 1996 and 2015. Methods Conventional microbiological methods and molecular techniques were used to study 114 isolates of N. meningitidis and 12 clinical samples without isolation (confirmed by polymerase chain reaction), provided by various sentinel centers and collaborating centers in Paraguay. Results The most affected age group was children under 1 year (19.0%), followed by 1-5-year-olds (17.5%). The highest percentage of strains was isolated in meningitis cases (81.7%) and serogroup B was found in 60.3% of cases. The most frequent phenotypes were B:4:p1.14 (16.0%), B:15:p1.5, C:nt:nst, and W:nt:p1.2 (10.5%), respectively. The prevalent clonal complexes were: ST-11/ET37 complex, 29.6% (8/27), predominantly serogroup W (6/8); ST-35 complex, 18.5% (5/27), in serogroup B (4/4); and ST-32/ET5 complex, 14.8% (4/16), in serogroup B (5/5). Conclusions Meningococcal meningitis is relatively uncommon in Paraguay. Distribution analysis showed that serogroup B is the most common and that the number of cases of meningococcal disease caused by serogroups C and W increased in the last two years. The identified clonal complexes were correlated with those found in the Southern Cone region. Due to the high virulence of N. meningitidis, its surveillance should be a strategic priority of national and regional public health systems to prevent epidemic outbreaks and support public health decision-making.

RESUMO Objetivo Descrever as características fenotípicas e genotípicas de cepas de Neisseria meningitidis isoladas de casos de doença meningocócica no Paraguai entre 1996 e 2015. Métodos Foram estudados por métodos microbiológicos convencionais e técnicas moleculares 114 isolados de N. meningitidis e 12 amostras clínicas sem isolamento confirmadas por reação em cadeia da polimerase (PCR) enviados por diferentes centros-sentinela e centros colaboradores do Paraguai. Resultados A faixa etária mais afetada foi a de crianças menores de 1 ano (19,0%) e crianças de 1 a 5 anos (17,5%). Uma maior porcentagem de cepas foi isolada de casos de meningite (81,7%) e o sorogrupo B foi identificado em 60,3% dos casos. Os fenótipos mais comuns foram B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST e W:NT:P1.2 (10,5%), respectivamente. Os complexos clonais mais prevalentes foram o complexo ST-11/ET37 (29,6%, 8/27) com predomínio no sorogrupo W (6/8), complexo ST-35 (18,5%, 5/27) no sorogrupo B (4/4) e complexo ST-32/ET5 (14,8%, 4/16) no sorogrupo B (5/5). Conclusões A doença meningocócica é relativamente pouco comum no Paraguai. A análise da distribuição dos sorogrupos demonstrou que o sorogrupo B é o mais prevalente e, nos últimos dois anos, ouve um aumento nos casos de doença meningocócica pelos sorogrupos C e W. Os complexos clonais encontrados se correlacionam com os achados na região do Cone Sul. Devido à alta virulência da N. meningitidis, a vigilância deste agente deve ser uma prioridade estratégica dos sistemas de saúde pública nacionais e regionais para prevenir surtos epidêmicos e subsidiar a tomada de decisão em saúde pública.

Are there any risk factors for developing complications with the use of retrievable vena cava filters in orthopaedic surgery?

BACKGROUND: In high-risk patients, common prophylaxis may be insufficient to prevent thromboembolic events after orthopaedic procedures. In this scenario, a retrievable vena cava filter (VCF) could be considered as an alternative, although it's use remains controversial. Therefore, we asked: (1) what is the overall mechanical complication rate associated with the use of retrievable VCFs in orthopaedic surgery?, (2) what is the association with thromboembolic disease (TED) recurrence, post-thrombotic syndrome and/or major bleeding according to different surgical characteristics?, (3) What is the overall mortality rate attributed to VCF use? METHODS: We retrospectively analyzed a cohort of 68 patients who underwent orthopaedic surgery with a previous diagnosis of TED, in whom a retrievable VCF was placed. Permanent filters were excluded. We studied the filter's mechanical complications and considered as possible outcomes death and 3 hematologic complications: TED recurrence, post-thrombotic syndrome and major bleeding. To estimate association with risk factors, we subclassified surgeries into 5 groups: 1, arthroplasty/non-arthroplasty; 2, primary/revision; 3, elective/urgent; 4, oncologic/non-oncologic; 5, preoperative/postoperative filter. RESULTS: Mechanical complications were 16% and required a filter revision. Sixty-four percent of the revised VCFs developed a mechanical failure and could not be retrieved. Overall prevalence of TED recurrence, post-thrombotic syndrome and hemorrhage was 33%, 15% and 4.5%, respectively. Spinal surgeries were a risk factor for developing TED recurrences.  Only 4% of patients died of a TED recurrence. CONCLUSIONS: Orthopaedic procedures had a high risk of mechanical and hematologic complications after using a retrievable VCF. However, mortality was low due to these complications.

A Dynamic Definition of Acute Kidney Injury Does not Improve Prognosis Assessment in Acutely Decompensated Patients with Cirrhosis.

BACKGROUND/OBJECTIVES: to compare the prognostic accuracy for 28 and 90-day transplant-free mortality of a modified CLIF-SOFA score (including a dynamic definition of acute kidney injury) with that of the classic CLIF-SOFA score and KDIGO score for acute kidney injury in patients with acute decompensation of cirrhosis. METHODS: A retrospective analysis of all admissions of acutely decompensated patients with cirrhosis was carried out from January 2012 to December 2014. Classic and modified CLIF-SOFA scores were analyzed, as well as acute kidney injury diagnosis using the KDIGO score regarding their accuracy for 28- and 90-day transplant free mortality prediction. RESULTS: 108 admissions were analyzed. Acute kidney injury diagnosis was met in 37 (34%) patients. Acute-on-chronic liver failure was diagnosed in 59 (55%) patients using the classic CLIF-SOFA score; and in 64 (59%) patients using the modified CLIF-SOFA score. Both CLIF-SOFA scores were highly effective in predicting 28-day transplant-free mortality (AUCROC 0.93 and 0.92, p = 0.34) as well as 90-day transplant-free mortality (AUCROC 0.79 and 0.78, p = 0.78). Acute kidney injury diagnosis had significantly lower accuracy in mortality assessment (28 and 90-day transplant free mortality AUCROC 0.67 [p = 0.002] and 0.63 [p = 0.02]). CONCLUSIONS: To our knowledge, this is the first evidence of the limited impact of modifying the fixed kidney injury definition currently used for acute-on-chronic liver failure.

Identification of New Tumor Suppressor Genes in Triple-Negative Breast Cancer.

Although genomic sequencing has provided a better understating of the genetic landmarks in triple-negative breast cancer (TNBC), functional validation of candidate cancer genes (CCG) remains unsolved. In this study, we used a transposon mutagenesis strategy based on a two-step sleeping beauty (SB) forward genetic screen to identify and validate new tumor suppressors (TS) in this disease. We generated 120 siRNAs targeting 40 SB-identified candidate breast cancer TS genes and used them to downregulate expression of these genes in four human TNBC cell lines. Among CCG, whose SB-mediated genetic mutation resulted in increased cellular proliferation in all cell lines tested, the genes ADNP, AP2B1, TOMM70A, and ZNF326 showed TS activity in tumor xenograft studies. Subsequent studies showed that ZNF326 regulated expression of multiple epithelial-mesenchymal transition and cancer stem cell (CSC) pathway genes. It also modulated expression of TS genes involved in the regulation of migration and cellular invasion and was a direct transcriptional activator of genes that regulate CSC self-renewal. ZNF326 expression associated with TNBC patient survival, with ZNF326 protein levels showing a marked reduction in TNBC. Our validation of several new TS genes in TNBC demonstrate the utility of two-step forward genetic screens in mice and offer an invaluable tool to identify novel candidate therapeutic pathways and targets. Cancer Res; 77(15); 4089-101. ©2017 AACR.

Calibración y validación de la Razón Internacional Normatizada (RIN) en pacientes cirróticos / Calibration and Validation of the International Normalized Ratio (INR) in Cirrhotic Patients

El MELD es un modelo pronóstico de score matemático usado para priorizar los pacientes en lista de espera para trasplante hepático, incluye resultados de creatinina, bilirrubina y tiempo de protrombina expresado como RIN. La disparidad en el score MELD como resultado de la variabilidad interlaboratorio de los componentes de la formula nos lleva a cuestionar la validez del mismo como herramienta de medición objetiva para la locación del órgano. El motivo de utilizar el MELD se basa en la presunción que el score debería ser igual en distintos lugares, si los métodos utilizados para medir las distintas variables llegaran al mismo resultado numérico. La evidencia muestra que la metodología utilizada para las mediciones puede influenciar en el cálculo del MELD identificando al RIN como la variable más importante. Esta variabilidad está dada por la distinta procedencia biológica de las tromboplastinas y de su ISI el cual refleja la capacidad de respuesta de la tromboplastina a la disminución de los factores de coagulación dependientes de la vitamina k. El RIN estandariza el tiempo de protrombina durante la anticoagulación oral, su uso se extendió para estandarizar el tiempo de protrombina en la enfermedad hepática y se incluyó en los modelos pronósticos como el MELD. Sin embrago los mecanismos de prolongación del tiempo de protrombina en la enfermedad hepática difieren de aquellos implicados en la anticoagulación oral y las tromboplastinas difieren en su sensibilidad para detectar las variaciones en los diferentes mecanismos. Tripodi y Velez han propuesto que los valores de ISI para las distintas tromboplastinas deberían ser calculados con plasmas de pacientes cirróticos y así calcular el RIN hepático lo que resolvería la variabilidad relacionada al RIN en el cálculo del MELD.

MELD is a prognostic model from amathematical score used to prioritize patients on a waiting list for liver transplantation and includes creatinine, bilirubin and prothrombin time expressed as an INR. The disparity in the MELD score, as a result of interlaboratory variability of the formula components, has lead us to question its validity as an objective measuring tool to localize the organ. The reason to use the MELD is based on the assumption that the score should be the same through different places, if methods used to measure the different variables reached the same numerical results. The evidence shows that the methods used in measuring can affect the MELD assessment by identifying the INR as the most important variable. This variability is caused by the different biological origin of the thromboplastins and their ISI, which reflects the thromboplastin's capacity of response to the decrease of vitamin K-dependent coagulation factors. The INR standardizes the prothrombin time during oral coagulation; its use was extended to standardize the prothrombin time in liver disease and was included in prognostic models like MELD. However, the mechanisms to extend the prothrombin time in liver disease are different from those involved in oral anticoagulation and the sensitivity of thromboplastins differ when detecting the variations in the different mechanisms. Tripodi and Velez have proposed that the ISI values for the different thromboplastins should be calculated on the basis of plasma from cirrhotic patients and thus the liver IRN should be calculated as well, which would resolve the variability associated to the IRN in calculating the MELD.

Intracellular targeting of annexin A2 inhibits tumor cell adhesion, migration, and in vivo grafting.

Cytoskeletal-associated proteins play an active role in coordinating the adhesion and migration machinery in cancer progression. To identify functional protein networks and potential inhibitors, we screened an internalizing phage (iPhage) display library in tumor cells, and selected LGRFYAASG as a cytosol-targeting peptide. By affinity purification and mass spectrometry, intracellular annexin A2 was identified as the corresponding binding protein. Consistently, annexin A2 and a cell-internalizing, penetratin-fused version of the selected peptide (LGRFYAASG-pen) co-localized and specifically accumulated in the cytoplasm at the cell edges and cell-cell contacts. Functionally, tumor cells incubated with LGRFYAASG-pen showed disruption of filamentous actin, focal adhesions and caveolae-mediated membrane trafficking, resulting in impaired cell adhesion and migration in vitro. These effects were paralleled by a decrease in the phosphorylation of both focal adhesion kinase (Fak) and protein kinase B (Akt). Likewise, tumor cells pretreated with LGRFYAASG-pen exhibited an impaired capacity to colonize the lungs in vivo in several mouse models. Together, our findings demonstrate an unrecognized functional link between intracellular annexin A2 and tumor cell adhesion, migration and in vivo grafting. Moreover, this work uncovers a new peptide motif that binds to and inhibits intracellular annexin A2 as a candidate therapeutic lead for potential translation into clinical applications.