An Aptamer against MNK1 for Non-Small Cell Lung Cancer Treatment.

Lung cancer is the leading cause of cancer-related death worldwide. Its late diagnosis and consequently poor survival make necessary the search for new therapeutic targets. The mitogen-activated protein kinase (MAPK)-interacting kinase 1 (MNK1) is overexpressed in lung cancer and correlates with poor overall survival in non-small cell lung cancer (NSCLC) patients. The previously identified and optimized aptamer from our laboratory against MNK1, apMNKQ2, showed promising results as an antitumor drug in breast cancer in vitro and in vivo. Thus, the present study shows the antitumor potential of apMNKQ2 in another type of cancer where MNK1 plays a significant role, such as NSCLC. The effect of apMNKQ2 in lung cancer was studied with viability, toxicity, clonogenic, migration, invasion, and in vivo efficacy assays. Our results show that apMNKQ2 arrests the cell cycle and reduces viability, colony formation, migration, invasion, and epithelial-mesenchymal transition (EMT) processes in NSCLC cells. In addition, apMNKQ2 reduces tumor growth in an A549-cell line NSCLC xenograft model. In summary, targeting MNK1 with a specific aptamer may provide an innovative strategy for lung cancer treatment.

An optimized MNK1b aptamer, apMNKQ2, and its potential use as a therapeutic agent in breast cancer.

Breast cancer is the most commonly diagnosed and leading cause of cancer death among women worldwide. Mitogen-activated protein kinase-interacting kinases (MNKs) promote the expression of several oncogenic proteins and are overexpressed in several types of cancer. In human cells, there are four isoforms of MNKs. The truncated isoform MNK1b, first described in our laboratory, has a higher basal activity and is constitutively active. Aptamers are emerging in recent years as potential therapeutic agents that show significant advantages over drugs of other nature. We have previously obtained and characterized a highly specific aptamer against MNK1b, named apMNK2F, with a dissociation constant in the nanomolar range, which produces significant inhibition of proliferation, migration, and colony formation in breast cancer cells. Furthermore, its sequence analysis predicted two G-quadruplex structures. In this work, we show the optimization process of the aptamer to reduce its size, improving its stability. The obtained aptamer, named apMNKQ2, is able to inhibit proliferation, colony formation, migration, and invasion in breast cancer cells. In murine models of breast cancer, apMNKQ2 has demonstrated its efficacy in reducing tumor volume and the number of metastases. In conclusion, apMNKQ2 could be used as an anti-tumor drug in the future.

Direct seeding as a recruitment alternative for the threatened tropical palm Syagrus coronata (Mart.) Beccari in Brazilian dry forest.

Habitat loss reduces biodiversity and threatens ecological services. The use of techniques for vegetation restoration such as direct seeding seems promising, mainly because it reduces costs and labor. The aim of this study was to investigate which are the main ecological filters that can inhibit the success of direct seeding, using Syagrus coronata (Mart.) Beccari (Arecaceae), a species that has great ecological and extractive importance for the Brazilian semi-arid region, and is found in habitats with different management intensities. The following hypotheses were tested: (1) managed habitat (pasture) and natural habitat (caatinga vegetation) show differences in seed fate; and (2) defleshed fruits will have greater seed germination success compared to not defleshed. Seed fate was evaluated for two consecutive years, year I (2016-2017) and year II (2017-2018). Within these years, 800 mature fruits were buried along 300 m transects, half with defleshing treatment, and monitored during 240 days. The highest seed survival occurred in year II, this result may be associated with higher rainfall recorded during the experiment, considering that there was no difference in seed germination between habitats. Defleshing facilitated germination and reduced insect predation, likely by removing invertebrate larvae before they have reached the seed. Desiccation was the major cause of mortality in the entire study and was higher in the managed habitat. Water insufficiency may have caused the deactivation of antioxidant mechanisms and affected the embryo. Although defleshing may favor germination, this procedure would not be indicated for pastures as it favors desiccation. The study showed how habitat management and fruit defleshing can affect seed fate. More studies on ecological relationships need to be carried out to increase understanding of how habitat modification affects the functioning of arid ecosystems.

The impact of vaccination on the evolution of COVID-19 in Portugal.

In this work we use simple mathematical models to study the impact of vaccination against COVID-19 in Portugal. First, we fit a SEIR type model without vaccination to the Portuguese data on confirmed cases of COVID-19 by the date of symptom onset, from the beginning of the epidemic until the 23rd January of 2021, to estimate changes in the transmission intensity. Then, by including vaccination in the model we develop different scenarios for the fade-out of the non pharmacological intervention (NPIs) as vaccine coverage increases in the population according to Portuguese vaccination goals. We include a feedback function to mimic the implementation and relaxation of NPIs, according to some disease incidence thresholds defined by the Portuguese health authorities.

DNA Aptamers against Vaccinia-Related Kinase (VRK) 1 Block Proliferation in MCF7 Breast Cancer Cells.

Vaccinia-related kinase (VRK) 1 is a serin/threonine kinase that plays an important role in DNA damage response (DDR), phosphorylating some proteins involved in this process such as 53BP1, NBS1 or H2AX, and in the cell cycle progression. In addition, VRK1 is overexpressed in many cancer types and its correlation with poor prognosis has been determined, showing VRK1 as a new therapeutic target in oncology. Using in vitro selection, high-affinity DNA aptamers to VRK1 were selected from a library of ssDNA. Selection was monitored using the enzyme-linked oligonucleotide assay (ELONA), and the selected aptamer population was cloned and sequenced. Three aptamers were selected and characterized. These aptamers recognized the protein kinase VRK1 with an affinity in the nanomolar range and showed a high sensibility. Moreover, the treatment of the MCF7 breast cell line with these aptamers resulted in a decrease in cyclin D1 levels, and an inhibition of cell cycle progression by G1 phase arrest, which induced apoptosis in cells. These results suggest that these aptamers are specific inhibitors of VRK1 that might be developed as potential drugs for the treatment of cancer.

Epidemic growth and Griffiths effects on an emergent network of excited atoms.

Whether it be physical, biological or social processes, complex systems exhibit dynamics that are exceedingly difficult to understand or predict from underlying principles. Here we report a striking correspondence between the excitation dynamics of a laser driven gas of Rydberg atoms and the spreading of diseases, which in turn opens up a controllable platform for studying non-equilibrium dynamics on complex networks. The competition between facilitated excitation and spontaneous decay results in sub-exponential growth of the excitation number, which is empirically observed in real epidemics. Based on this we develop a quantitative microscopic susceptible-infected-susceptible model which links the growth and final excitation density to the dynamics of an emergent heterogeneous network and rare active region effects associated to an extended Griffiths phase. This provides physical insights into the nature of non-equilibrium criticality in driven many-body systems and the mechanisms leading to non-universal power-laws in the dynamics of complex systems.

Analysis of potentially inappropriate medications prescribed to older patients in a hospital setting / Análisis de medicamentos potencialmente inapropiados recetados a personas mayores pacientes en un entorno hospitalario

INTRODUCTION: An ever increasing number of elderly live with multiple chronic diseases and take several drugs concomitantly. The elderly are subjected to a variety of pharmacodynamic and pharmacokinetic alterations that change the responsiveness of drugs, which makes them potentially inappropriate to use in older people. The aim of this study is to quantitatively assess the prescription pattern of potentially inappropriate medications (PIMs) in different inward clinical services of University Hospital Center of Cova da Beira, Covilhã, Portugal. METHODS: We searched the hospital's computerized system database for every patient admitted in previously selected clinical services, from January 1st to June 30th 2018. Patients less than 65 years old were excluded. Data regarding the patients' hospital ID number, their age, prescribed drugs during admission, prescribed dose, frequency, medications' start date and their respective end date were anonymously collected. The patients' medications prescribed during the admission period were assessed, and the PIMs were identified according to Beers Criteria 2015. RESULTS: The benzodiazepines were the most frequently prescribed potentially inappropriate therapeutic drug class in the four clinical services studied, accounting for 29.97% in Medicine 2, 39.96% in Cardiology, 30.21% in Medicine 1 and 41.10% in Pneumology of the total of prescribed PIMs. The antipsychotics were the next most prescribed potentially inappropriate therapeutic drug classes. Intestinal motility modifiers, namely metoclopramide, also had a significant expression. CONCLUSIONS: Reconciliation tools such as the Beers Criteria are useful to identify inappropriate prescribing during the pharmaceutical validation of prescription. Further studies will provide more insight into the impact of the pharmacist's intervention

INTRODUCCIÓN: Un número cada vez mayor de ancianos vive con múltiples enfermedades crónicas y toma varios medicamentos concomitantementes. Los ancianos presentan una variedad de alteraciones farmacodinámicas y farmacocinéticas que alteran la respuesta de los fármacos, lo que los hace potencialmente inapropiados para su uso en estos pacientes. El objetivo de este estudio es evaluar cuantitativamente el patrón de prescripción de medicamentos potencialmente inapropiados (PIMs) en diferentes servicios clínicos de hospitalización en el Centro Hospitalario Universitario de Cova da Beira, Covilha, Portugal. MÉTODOS: Se realizó una investigación en la base de datos del sistema informático del hospital para todos los pacientes internados en servicios clínicos previamente seleccionados, del 1 de enero al 30 de junio de 2018. Se excluyeron a los pacientes menores de 65 años. Los datos referentes al número del proceso clínico del paciente, edad, medicamentos prescritos en el momento de la admisión en el hospital, dosis prescrita, frecuencia, fecha de inicio de los medicamentos y sus respectivas fechas finales fueron recolectados anónimamente. Se evaluaron los medicamentos prescritos de los pacientes durante el período de internamiento, y los PIMs se identificaron de acuerdo con los criterios de Beers de 2015. RESULTADOS: Las benzodiazepinas fueron la clase terapéutica medicamentosa potencialmente inapropiada prescrita con mayor frecuencia en los cuatro servicios clínicos estudiados, siendo el 29,97% en la Medicina 2, el 39,96% en la Cardiología, el 30,21% en la Medicina 1 y el 41,10% en la Neumología del total de PIMs prescritos. Los antipsicóticos fueron la segunda clase más prescrita de PIMs. Los modificadores de la motilidad intestinal, en particular la metoclopramida, también tuvieron una expresión significativa. CONCLUSIONES: Las herramientas de reconciliación, como los criterios de Beers, son útiles para identificar prescripciones inadecuadas durante la validación farmacéutica de la prescripción. Otros estudios proporcionarán más información sobre el impacto de la intervención del farmacéutico

The prevention of doping and the improper use of drugs and food supplements in sports and physical activities: a survey on the activity of the prevention departments of Italian local health authorities.

INTRODUCTION: Doping is an important public health problem widespread not only among elite athletes, but also among amateur and recreational athletes and the general population. In Italy the introduction of doping prevention within the Essential Levels of Care (LEA) with the DPCM 12/1/2017 represents a crucial step towards the implementation of education and health promotion interventions. In this context, the Departments of Prevention (DP) of the Local Health Authorities (LHA) have to play a fundamental role, becoming the cultural and operational reference on this issue. As part of the "Doping prevention: development of a permanent educational tool coordinated by the National Health Service Prevention Departments" project, funded by the Italian Ministry of Health, a survey was conducted on the activities carried out by the DP regarding doping prevention and improper use and abuse of drugs and food supplements in sports and physical activities, as a basis for the harmonization of organizational structures and prevention programs and the creation of a collaboration network at a regional and national level. METHODS: A semi-structured questionnaire consisting of 11 questions, prepared on an electronic platform, was sent to the DP of all the Italian LHA. RESULTS: A total of 38 DP out of 131 (29%) completed the questionnaire, with representation from all regions. 42.1% of DP carried out or are still running programs for the prevention of doping, a percentage that decreases to 27% considering the programs for the prevention of misuse and abuse of drugs and food supplements in sports and in physical activities; in less than half of the DP, 37.5% and 41.7%, respectively, dedicated funds have been allocated. The professionals most involved in prevention of doping are the Specialists in Sport Medicine (81.3%) followed by Specialists in Hygiene (43.8%) and Psychologists (37.5%), while Health Care Assistants (50%) are the professionals most involved in the prevention of the improper use of drugs and food supplements, followed by Specialists in Hygiene and Specialists in Sport Medicine (40%). Most of the DP (71.9%) believe that the introduction of programs to prevent and counteract doping in the LEA will have repercussions on their approach against doping. CONCLUSIONS: The survey, although conducted on a limited sample, has provided an important framework relating to programs for the prevention of doping and the misuse and abuse of drugs and food supplements in sports and in the physical activities carried out by DP. A remarkable heterogeneity has been highlighted, both at national and regional level. It is urgent to provide DP with homogeneous and effective organizational models and adequate operational tools, paying particular attention to the training of all the professionals involved. It is also essential to implement permanent monitoring tools.

Penile lengthening with porcine small intestinal submucosa grafting in Peyronie's disease treatment: long-term surgical outcomes, patients' satisfaction and dissatisfaction predictors.

BACKGROUND: Peyronie's Disease (PD) is an acquired connective tissue disorder that often leads to penile curvature (PC) and sexual dysfunction. Penile lengthening procedures (PLP) with four-layered porcine small intestinal submucosa graft (Surgisis® ES; Cook) have been widely used in patients with severe PC when erectile function is preserved. However, complications such as erectile dysfunction (ED) may limit treatment satisfaction. OBJECTIVES: Focusing on patients perspective, our study aims to evaluate longterm patient-reported outcomes, satisfaction, and dissatisfaction predictors after PLP. MATERIAL AND METHODS: This prospective study included 32 patients affected by PD with severe PC submitted to PLP with Surgisis® ES between 2011 and 2014. All patients were submitted to a standardized protocol with regular clinical evaluation at 3, 6 and 12 months, and yearly thereafter. After the third year follow-up, IIEF-5, modified EDITS and an additional non-validated questionnaire were completed. RESULTS: Concerning the surgical procedure, the mean tunical defect area (TDA) was 15.9 ± 6.9 cm2 . The mean follow-up time were 49.6 ± 12.7 months and there was a significant increase in stretched penile length (p = 0.01). Postoperative erectile function as assessed by IIEF-5 was positively correlated with overall treatment satisfaction evaluated through EDITS (p = 0.01). TDA was negatively correlated with postoperative IIEF-5 (R = -0.56, p < 0.001). TDA ≥ 14.375 cm2 can predict ED with 76.9% sensibility and 58.3% specificity. Moreover, TDA ≥ 21.875 cm2 can predict clinically significant ED (IIEF-5 score ≤17) with 80% sensibility and 95.2% specificity. Patient-reported longterm complications were 65.6% decreased penile length, 56.5% diminished rigidity and 25% curvature recurrence. CONCLUSION: PLP using a Surgisis® ES is a valuable surgical option for the treatment of PD with severe PC. Although it results in high rates of long-term patient-reported overall satisfaction, possible outcomes such as postoperative ED must be acknowledged. TDA is a strong predictor for postoperative ED and should be considered in clinical practice to classify patients in low- or high-risk for postoperative ED.

Immunization and media coverage in Italy: an eleven-year analysis (2007-17).

Immunization polices in Italy has recently reached important milestones, including the approval of the National Immunization Prevention Plan and of a new law mandatory immunization; this stimulating a lively debate at the scientific, political and societal-level, reflected on the media. We applied a model previously published to quantitatively and qualitatively assess media coverage on vaccines and immunization-related topics on the most read Italian newspaper, "Corriere della Sera", over an 11-year study period (2007-2017). We retrieved relevant key words and articles, reported on included articles' topic, position, approach to immunization and on other selected indicators' summary statistics, temporal trends and correspondence with key epidemiological and policy events. Over the study period the quote "vaccin*" was mentioned on average 325 times per year; with an increase of 150% after the approval of the new law on mandatory immunization in 2017. In the same year, on average, two first-page articles per week were published on the topic. We report a clear association between key events (i.e the H1N1 influenza pandemic, the "Fluad case", the approval of the new law on mandatory immunization) and their media coverage. Overall, 84% of articles had a positive attitude towards immunization, this share decreasing to 79% when only considering articles published after the approval of the law on mandatory vaccination. Media play a crucial role in channelling health-related information and significantly influence health behaviours. We urge public institutions, health authorities and the scientific community not to underestimate the opportunity to monitor media coverage on key healthcare topics and to convey evidence-based health education messages through the media.

Increased expression of MNK1b, the spliced isoform of MNK1, predicts poor prognosis and is associated with triple-negative breast cancer.

MAP kinase interacting kinases (MNKs) modulate the function of oncogene eukaryotic initiation factor 4E (eIF4E) through phosphorylation, which is necessary for oncogenic transformation. MNK1 gives rise to two mRNAs and thus two MNK1 isoforms, named MNK1a and MNK1b. MNK1b, the splice variant of human MNK1a, is constitutively active and independent of upstream MAP kinases. In this study, we have analyzed the expression of both MNK1 isoforms in 69 breast tumor samples and its association with clinicopathologic/prognostic characteristics of breast cancer. MNK1a and MNK1b expression was significantly increased in tumors relative to the corresponding adjacent normal tissue (p < 0.001). In addition, MNK1b overexpression was found in most of the triple-negative tumors and was associated with a shorter overall and disease-free survival time. Overexpression of MNK1b in MDA-MB-231 cells induced an increase in the expression of the MCL1 antiapoptotic protein and promoted proliferation, invasion and colony formation. In conclusion, a high expression level of MNK1b protein could be used as a marker of poor prognosis in breast cancer patients and it could be a therapeutic target in triple-negative tumors.

Changes in serum interleukin-8 (IL-8) levels reflect and predict response to anti-PD-1 treatment in melanoma and non-small-cell lung cancer patients.

BACKGROUND: Surrogate biomarkers of efficacy are needed for anti-PD1/PD-L1 therapy, given the existence of delayed responses and pseudo-progressions. We evaluated changes in serum IL-8 levels as a biomarker of response to anti-PD-1 blockade in melanoma and non-small-cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: Metastatic melanoma and NSCLC patients treated with nivolumab or pembrolizumab alone or nivolumab plus ipilimumab were studied. Serum was collected at baseline; at 2-4 weeks after the first dose; and at the time-points of response evaluation. Serum IL-8 levels were determined by sandwich ELISA. Changes in serum IL-8 levels were compared with the Wilcoxon test and their strength of association with response was assessed with the Mann-Whitney test. Accuracy of changes in IL-8 levels to predict response was estimated using receiver operation characteristics curves. RESULTS: Twenty-nine melanoma patients treated with nivolumab or pembrolizumab were studied. In responding patients, serum IL-8 levels significantly decreased between baseline and best response (P <0.001), and significantly increased upon progression (P =  0.004). In non-responders, IL-8 levels significantly increased between baseline and progression (P =  0.013). Early changes in serum IL-8 levels (2-4 weeks after treatment initiation) were strongly associated with response (P <0.001). These observations were validated in 19 NSCLC patients treated with nivolumab or pembrolizumab (P =  0.001), and in 15 melanoma patients treated with nivolumab plus ipilimumab (P <0.001). Early decreases in serum IL-8 levels were associated with longer overall survival in melanoma (P =  0.001) and NSCLC (P =  0.015) patients. Serum IL-8 levels also correctly reflected true response in three cancer patients presenting pseudoprogression. CONCLUSIONS: Changes in serum IL-8 levels could be used to monitor and predict clinical benefit from immune checkpoint blockade in melanoma and NSCLC patients.

Inhibition of Influenza Virus Replication by DNA Aptamers Targeting a Cellular Component of Translation Initiation.

The genetic diversity of the influenza virus hinders the use of broad spectrum antiviral drugs and favors the appearance of resistant strains. Single-stranded DNA aptamers represent an innovative approach with potential application as antiviral compounds. The mRNAs of influenza virus possess a 5'cap structure and a 3'poly(A) tail that makes them structurally indistinguishable from cellular mRNAs. However, selective translation of viral mRNAs occurs in infected cells through a discriminatory mechanism, whereby viral polymerase and NS1 interact with components of the translation initiation complex, such as the eIF4GI and PABP1 proteins. We have studied the potential of two specific aptamers that recognize PABP1 (ApPABP7 and ApPABP11) to act as anti-influenza drugs. Both aptamers reduce viral genome expression and the production of infective influenza virus particles. The interaction of viral polymerase with the eIF4GI translation initiation factor is hindered by transfection of infected cells with both PABP1 aptamers, and ApPABP11 also inhibits the association of NS1 with PABP1 and eIF4GI. These results indicate that aptamers targeting the host factors that interact with viral proteins may potentially have a broad therapeutic spectrum, reducing the appearance of escape mutants and resistant subtypes.

Characterization of MNK1b DNA Aptamers That Inhibit Proliferation in MDA-MB231 Breast Cancer Cells.

Elevated expression levels of eukaryotic initiation factor 4E (eIF4E) promote cancer development and progression. MAP kinase interacting kinases (MNKs) modulate the function of eIF4E through the phosphorylation that is necessary for oncogenic transformation. Therefore, pharmacologic MNK inhibitors may provide a nontoxic and effective anticancer strategy. MNK1b is a truncated isoform of MNK1a that is active in the absence of stimuli. Using in vitro selection, high-affinity DNA aptamers to MNK1b were selected from a library of ssDNA. Selection was monitored using the enzyme-linked oligonucleotide assay (ELONA), and the selected aptamer population was cloned and sequenced. Four groups of aptamers were identified, and the affinities of one representative for rMNK1b were determined using ELONA and quantitative polymerase chain reaction. Two aptamers, named apMNK2F and apMNK3R, had a lower Kd in the nmol/l range. The secondary structure of the selected aptamers was predicted using mFold, and the QGRS Mapper indicated the presence of potential G-quadruplex structures in both aptamers. The selected aptamers were highly specific against MNK1, showing higher affinity to MNK1b than to MNK1a. Interestingly, both aptamers were able to produce significant translation inhibition and prevent tumor cell proliferation and migration and colony formation in breast cancer cells. These results indicate that MNK1 aptamers have an attractive therapeutic potential.

Microbiological quality control of non-sterile compounded medicines prepared in a Portuguese hospital centre.

OBJECTIVES: This work aimed to evaluate the quality of non-sterile formulations compounded at Centro Hospitalar Cova da Beira (Covilhã, Portugal) immediately after preparation and up to the defined 'beyond-use date'. METHODS: Microbiological quality control tests were performed in accordance with monograph 5.1.4 of the European Pharmacopoeia 8.0. Samples of compounded products were collected from January to December 2014 after preparation and were analysed immediately and reanalysed after storage under the established conditions, for each preparation. RESULTS: In the test period, 392 preparations were analysed, corresponding to 24 different formulations (8 intermediate preparations, 11 oral solutions/suspensions and 5 topical preparations). All preparations were in accordance with the pharmacopoeia specifications immediately after preparation. However, for the formulations 'prednisolone oral solution (5 mg/mL)' and 'nitroglycerine and cinchocaine ointment (0.25%/0.5%)', the microbial counts of some batches exceeded the defined limits after storage up to the beyond-use date. CONCLUSIONS: These results show that the compounding practices implemented at this pharmacy department are able to ensure the microbiological quality of compounded products. This microbiological quality control methodology also allowed identification of the need to replace formulations shown not to be stable throughout the storage period. On the basis of these results, a monthly routine of microbiological control of a random sample of compounded medicines was established in order to ensure their quality and safety for use.

IL-1ß induces p62/SQSTM1 and represses androgen receptor expression in prostate cancer cells.

Chronic inflammation is associated with advanced prostate cancer (PCa), although the mechanisms governing inflammation-mediated PCa progression are not fully understood. PCa progresses to an androgen independent phenotype that is incurable. We previously showed that androgen independent, androgen receptor negative (AR(-) ) PCa cell lines have high p62/SQSTM1 levels required for cell survival. We also showed that factors in the HS-5 bone marrow stromal cell (BMSC) conditioned medium can upregulate p62 in AR(+) PCa cell lines, leading us to investigate AR expression under those growth conditions. In this paper, mRNA, protein, and subcellular analyses reveal that HS-5 BMSC conditioned medium represses AR mRNA, protein, and nuclear accumulation in the C4-2 PCa cell line. Using published gene expression data, we identify the inflammatory cytokine, IL-1ß, as a candidate BMSC paracrine factor to regulate AR expression and find that IL-1ß is sufficient to both repress AR and upregulate p62 in multiple PCa cell lines. Immunostaining demonstrates that, while the C4-2 population shows a primarily homogeneous response to factors in HS-5 BMSC conditioned medium, IL-1ß elicits a strikingly heterogeneous response; suggesting that there are other regulatory factors in the conditioned medium. Finally, while we observe concomitant AR loss and p62 upregulation in IL-1ß-treated C4-2 cells, silencing of AR or p62 suggests that IL-1ß regulates their protein accumulation through independent pathways. Taken together, these in vitro results suggest that IL-1ß can drive PCa progression in an inflammatory microenvironment through AR repression and p62 induction to promote the development and survival of androgen independent PCa.

Expression of the transmembrane mucins, MUC1, MUC4 and MUC16, in normal endometrium and in endometriosis.

STUDY QUESTION: Are the transmembrane mucins, MUC1, MUC4 and MUC16, differentially expressed in endometriosis compared with normal endometrium? SUMMARY ANSWER: This study revealed that transmembrane mucin expression does not vary significantly in normal endometrium during the menstrual cycle and is not altered in endometriosis relative to the epithelial marker, cytokeratin-18 (KRT18). WHAT IS KNOWN ALREADY: Increased serum levels of the transmembrane mucin fragments MUC1, MUC4 and MUC16 that normally dominate the apical surface of simple epithelia are found in several pathological conditions, including endometriosis. Altered mucin expression in gynecologic diseases may promote infertility or endometrial pathologies. STUDY DESIGN, SIZE, DURATION: This was a laboratory-based study of samples from 12 endometriosis patients as well as non-endometriosis control samples obtained from 31 patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: Total RNA was isolated from endometrial biopsies of ectopic and eutopic endometrium from women with endometriosis and control patients from different stages of the menstrual cycle. Quantitative (q)-RT-PCR analyses were performed for the mucins, MUC1, MUC4 and MUC16, relative to the epithelial marker, cytokeratin-18 (KRT18), or ß-actin (ACTB). Frozen sections from endometrial biopsies of proliferative and mid-secretory stage women with endometriosis were immunostained for MUC1, MUC4 and MUC16. MAIN RESULTS AND THE ROLE OF CHANCE: qRT-PCR analyses of MUC1 and MUC16 mRNA revealed that these mucins do not vary significantly during the menstrual cycle nor are they altered in women with endometriosis relative to the epithelial marker, KRT18. MUC4 mRNA is expressed at very low levels relative to MUC1 and MUC16 under all conditions. There was little difference in MUC1 and MUC16 expression between eutopic endometrial and ectopic endometriotic tissues. MUC4 expression also was not significantly higher in the ectopic endometriotic tissues. Immunostaining for all three mucins reveals robust expression of MUC1 and MUC16 at the apical surfaces of endometrial epithelia, but little to no staining for MUC4. LIMITATIONS, REASONS FOR CAUTION: qRT-PCR analysis was the main method used for mucin detection. Additional studies with stage III-IV endometriotic tissue would be useful to determine if changes in MUC1 and MUC16 expression occur, or if MUC4 expression increases, at later stages of endometriosis. WIDER IMPLICATIONS OF THE FINDINGS: We report a comprehensive comparative profile of the major transmembrane mucins, MUC1, MUC4 and MUC16, relative to the epithelial marker, KRT18, in normal cycling endometrium and in endometriosis, and indicate constitutive expression. Previous studies have profiled the expression of individual mucins relative to ß-actin and indicate accumulation in the luteal phase. Thus, these differences in interpretation appear to reflect the increased epithelial content of endometrium during the luteal phase. STUDY FUNDING: This study was supported by: NIH R01HD29963 to D.D.C.; NIH U54HD007495 to S.M.H.; and NIH R01HD067721 to S.L.Y. and B.A.L. The authors have no competing interests to declare.

Aplicación del modelo de la recuperación a la psicosis en la práctica clínica cotidiana. Una experiencia grupal de 2 años / Recovery model in psychosis. Use in dayly clinical practice. A two years group experience

Introducción: Las premisas teóricas de la recuperación cuestionan el concepto kraepeliniano de la psicosis. Asumir la reversibilidad de esta condición favorece la incorporación de tratamientos integrados con el fin de abordar de forma eficiente las distintas dimensiones que la conforman. Objetivos del estudio: Describir la puesta en marcha de intervenciones grupales desde el punto de vista de la recuperación en personas con psicosis en un dispositivo comunitario de la red pública. Confirmar la eficacia de dichas intervenciones identificando los factores clínicos mejorados. Analizar el efecto de las intervenciones terapéuticas prolongadas en la evolución clínica. Pacientes: 28 personas con diagnósticos incluidos en los códigos CIE-10 (F20-25, F31, F60.0 y F60.1), tradicionalmente relacionados con la psicosis. Método: Se describe la estructura y el contenido del grupo, que han sido adaptados al contexto de trabajo: Grupo abierto en cuanto a pacientes y profesionales, sesiones quincenales, incorporación de técnicas corporales, evolución hacia la ayuda mutua etc... Se evalúa la situación clínica mediante las escalas PANSS y GAF. Resultados: Se objetiva una mejoría significativa a nivel de los síntomas negativos y sociales que se mantiene e incrementa entre los pacientes que continúan en seguimiento a lo largo del tiempo. Discusión: Dichos hallazgos resultan especialmente valiosos dada la resistencia de las dimensiones emocionales y sociales a los abordajes exclusivamente farmacológicos. Se plantean líneas de trabajo futuras, las dificultades en la generalización de los logros obtenidos en el contexto terapéutico a las actividades de la vida cotidiana y los ensayos que se han puesto en marcha para vencer dichas trabas. Asimismo se considera la posibilidad de incluir otros instrumentos de evaluación de naturaleza cualitativa que incluyan la visión subjetiva de los pacientes en su propio proceso (AU)

Introduction: Theoretical background in recovery confronts the kraepelian concept of psychosis. Considering this condition as reversible in volves looking for more efficient therapies for some of the dimensions. Objectives: Description of the process to start a group with people with psychosis under the recovery paradigm in a public health team. Confirmation of the efficacy of group intervention and treated symptoms. Analysis of effect of long-term group intervention. Patients: This work is based on a recovery group therapy experience in a community mental health team from a public health service in Spain. 28 subjects were diagnosed of disorders included in ICD- 10: F20-25, F31, F60.0 and F60.1 related to psychosis. Method: The study describes structure and contents of the group and clinical assessment trough PANSS and GAF scales. Results: Negative and social symptoms remarkably improved in a progressive way trough the two years course. Discussion: These facts are considered especially worthy given their traditionally resistant condition to an isolated pharmacological assessment. Future lines of study are drawn. Difficulties in generalization of results in real framework are discussed. We are considering new measurement tools including subjective and qualitative instruments (AU)

Association of the HLA-B*52 allele with non-progression to AIDS in Brazilian HIV-1-infected individuals.

Several human leukocyte antigen (HLA) class I alleles are associated with the susceptibility to human immunodeficiency virus-1 (HIV-1) infection and/or AIDS progression. Of these, the HLA-B alleles are considered the strongest genetic determinant of disease outcome. We evaluated the influence of the HLA-B alleles on AIDS progression among HIV-1-positive individuals from Rio de Janeiro, Brazil, who were categorized as rapid progressors (RPs), typical progressors (TPs) or long-term non-progressors (LTNPs). In this study, significant differences in HLA-B allele frequencies were observed among the three progression groups for the B*48, B*49 and B*52 alleles. After controlling for other factors associated with AIDS progression, the presence of the B*52 allele was shown to be a significant protective factor (hazard ratio (HR) 0.49 (95% confidence interval (CI) 0.27-0.90) P<0.03). Although no direct association was observed between the presence of the B*27 or B*57 allele and the LTNP profile compared with the TP or RP groups, the adjusted model confirmed that these alleles are protective factors against AIDS progression (HR 0.62 (95% CI 0.38-0.99) P<0.05), as previously described. These data corroborate the existence of significant differences in HLA-B allele frequencies among the distinct AIDS progression profiles and further elucidate the role of HLA alleles in the outcome of HIV infections in diverse populations.

eIF4F complex disruption causes protein synthesis inhibition during hypoxia in nerve growth factor (NGF)-differentiated PC12 cells.

Poor oxygenation (hypoxia) influences important physiological and pathological situations, including development, ischemia, stroke and cancer. Hypoxia induces protein synthesis inhibition that is primarily regulated at the level of initiation step. This regulation generally takes place at two stages, the phosphorylation of the subunit α of the eukaryotic initiation factor (eIF) 2 and the inhibition of the eIF4F complex availability by dephosphorylation of the inhibitory protein 4E-BP1 (eukaryotic initiation factor 4E-binding protein 1). The contribution of each of them is mainly dependent of the extent of the oxygen deprivation. We have evaluated the regulation of hypoxia-induced translation inhibition in nerve growth factor (NGF)-differentiated PC12 cells subjected to a low oxygen concentration (0.1%) at several times. Our findings indicate that protein synthesis inhibition occurs primarily by the disruption of eIF4F complex through 4E-BP1 dephosphorylation, which is produced by the inhibition of the mammalian target of rapamycin (mTOR) activity via the activation of REDD1 (regulated in development and DNA damage 1) protein in a hypoxia-inducible factor 1 (HIF1)-dependent manner, as well as the translocation of eIF4E to the nucleus. In addition, this mechanism is reinforced by the increase in 4E-BP1 levels, mainly at prolonged times of hypoxia.