Search

Focused electron beam induced deposition (FEBID) allows the creation of nanoscale structures through dissociation of an organo-metallic precursor by electrons at the beam impact point. The deposition of Pd can be interesting for its catalytic behavior and ability to contact carbon based materials. Two precursors were investigated—Pd(hfac)2 and (Cp)Pd(allyl)—and two deposition methods: with and without an in situ oxygen purification process. The deposition parameters can be tuned for the Pd(hfac)2 precursor to provide a deposition with 23 ± 2 at.% of Pd and a main component of C at 51 ± 3 at.% and minor components of O and F. An in situ purification process using O2 was much faster than expected and improved the Pd content to up to >65 at.% while reducing the C to ∼20 at.%, and avoiding the oxidation of Pd. The resistivity was ∼100 μOhm · cm and compares favorably with a bulk value of 10 μOhm · cm. The (Cp)Pd(allyl) precursor is interesting because it does not release fluorine during the deposition and hence it does not etch a possible substrate. Its FEBID deposition had a composition of 26 ± 5 at.% of Pd with 74 ± 5 at.% of C. The O2 purification process can improve the Pd content up to ∼60 at.% while reducing C to <20 at.%, but also increasing the O content to 18 at%, which was released afterwards. The best resistivity was measured at ∼1000 μOhm · cm, although better values can be anticipated for longer post treatment times. [ABSTRACT FROM AUTHOR]

Highly pure metallic structures can be deposited by electron beam induced deposition and they have many important applications in different fields. The organo-metallic precursor is decomposed and deposited under the electron beam, and typically it is purified with post-irradiation in presence of O2. However, this approach limits the purification to the surface of the deposit. Therefore, ‘in situ’ purification during deposition using simultaneous flows of both O2 and precursor in parallel with two gas injector needles has been tested and verified. To simplify the practical arrangements, a special concentric nozzle has been designed allowing deposition and purification performed together in a single step. With this new device metallic structures with high purity can be obtained more easily, while there is no limit on the height of the structures within a practical time frame. In this work, we summarize the first results obtained for ‘in situ’ Au purification using this concentric nozzle, which is described in more detail, including flow simulations. The operational parameter space is explored in order to optimize the shape as well as the purity of the deposits, which are evaluated through scanning electron microscope and energy dispersive x-ray spectroscopy measurements, respectively. The observed variations are interpreted in relation to other variables, such as the deposition yield. The resistivity of purified lines is also measured, and the influence of additional post treatments as a last purification step is studied. [ABSTRACT FROM AUTHOR]

Objectives: Cushing's disease (CD) in the context of chronic kidney disease (CKD) is very rare. CKD causes physiological hypercortisolism making the diagnosis of CD extremely difficult. To report 3 females with CKD and CD and to outline the principles that may guide the diagnosis of CD in this context., Case Presentation: P1. A 12.3-year-old patient with CKD secondary to steroid-resistant nephrotic syndrome on hemodialysis (HD) and a medical history of aseptic meningitis. She was referred due to the incidental finding of pituitary macroadenoma. P2. A patient with CKD secondary to bilateral renal hypodysplasia in conservative treatment. At age 16.4 years, she had significant weight gain, purple-red stretch marks, galactorrhea, and menstrual irregularities. P3. A 15.3-year-old patient with CKD secondary to steroid resistant nephrotic syndrome in conservative treatment was referred for weight gain, secondary amenorrhea, and hypertension. In all patients, diagnosis of CD was confirmed by clinical and biochemical findings. P1 and P3 underwent transsphenoidal surgery, and in P2, transcranial surgery resection was performed. Histopathological examination revealed a corticotroph adenoma in P1 and P2, and in P3, immunohistochemistry demonstrated ACTH predominance. All patients achieved remission. P1 and P2 developed pituitary deficiencies., Conclusions: To the best of our knowledge, these are the first three reported cases of the diagnostic association of CD and CKD in children. In all cases, CS was clinically suspected and CD was confirmed through complementary exams. Given the current lack of clear diagnostic criteria for CD in CKD patients, a thorough clinical evaluation remains essential for guiding the diagnosis., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

Background: The COVID-19 pandemic has had an unprecedented impact in the global research production and has also increased research waste. Living evidence syntheses (LESs) seek to regularly update a body of evidence addressing a specific question. During the COVID-19 pandemic, the production and dissemination of LESs emerged as a cornerstone of the evidence infrastructure. This critical interpretive synthesis answers the questions: What constitutes an LES to support decision-making?; when should one be produced, updated, and discontinued?; and how should one be disseminated?, Methods: Searches included the Cochrane Library, EMBASE (Ovid), Health Systems Evidence, MEDLINE (Ovid), PubMed, and Web of Science up to 23 April 2024 and included articles that provide any insights on addressing the compass questions on LESs. Articles were selected and appraised, and their insights extracted. An interpretive and iterative coding process was used to identify relevant thematic categories and create a conceptual framework., Results: Among the 16,630 non-duplicate records identified, 208 publications proved eligible. Most were non-empirical articles, followed by actual LESs. Approximately one in three articles were published in response to the COVID-19 pandemic. The conceptual framework addresses six thematic categories: (1) what is an LES; (2) what methodological approaches facilitate LESs production; (3) when to produce an LES; (4) when to update an LES; (5) how to make available the findings of an LES; and (6) when to discontinue LES updates., Conclusion: LESs can play a critical role in reducing research waste and ensuring alignment with advisory and decision-making processes. This critical interpretive synthesis provides relevant insights on how to better organize the global evidence architecture to support their production., Trial Registration: PROSPERO registration: CRD42021241875., (© 2024. The Author(s).)

Background: Globally, a growing number of calls to formalize and strengthen evidence-support systems have been released, all of which emphasize the importance of evidence-informed decision making. To achieve this, it is critical that evidence producers and decision-makers interact, and that decision-makers' evidence needs can be efficiently translated into questions to which evidence producers can respond. This paper aims to create a taxonomy of demand-driven questions for use by evidence producers, intermediaries (i.e., people working in between researchers and decision-makers) and decision-makers., Methods: We conducted a global cross-sectional survey of units providing some type of evidence support at the explicit request of decision-makers. Unit representatives were invited to answer an online questionnaire where they were asked to provide a list of the questions that they have addressed through their evidence-support mechanism. Descriptive analyses were used to analyze the survey responses, while the questions collected from each unit were iteratively analyzed to create a mutually exclusive and collectively exhaustive list of types of questions that can be answered with some form of evidence., Results: Twenty-nine individuals completed the questionnaire, and more than 250 submitted questions were analysed to create a taxonomy of 41 different types of demand-driven questions. These 41 questions were organized by the goal to be achieved, and the goals were grouped in the four decision-making stages (i) clarifying a societal problem, its causes and potential impacts; (ii) finding and selecting options to address a problem; (iii) implementing or scaling-up an option; and (iv) monitoring implementation and evaluating impacts., Conclusion: The mutually exclusive and collectively exhaustive list of demand-driven questions will help decision-makers (to ask and prioritize questions), evidence producers (to organize and present their work), and evidence-intermediaries (to connect evidence needs with evidence supply)., (© 2024. The Author(s).)

Research evidence can play an important role in each stage of decision-making, evidence-support systems play a key role in aligning the demand for and supply of evidence. This paper provides guidance on what type of study designs most suitably address questions asked by decision-makers. This study used a two-round online Delphi approach, including methodological experts in different areas, disciplines, and geographic locations. Participants prioritized study designs for each of 40 different types of question, with a Kendall's W greater than 0.6 and reaching statistical significance (p<0.05) considered as a consensus. For each type of question, we sorted the final rankings based on their median ranks and interquartile ranges, and listed the four study designs with the highest median ranks. Participants provided 29 answers in the two rounds of the Delphi, and reached a consensus for 28 (out of the 40) questions (eight in the first round and 20 in the second). Participants achieved a consensus for 8 of 15 questions in stage I (clarifying a societal problem, its causes, and potential impacts), 12 of 13 in stage II (finding options to address a problem) and four of six in each of stages III (implementing or scaling-up an option) and IV (monitoring implementation and evaluating impact). This paper provides guidance on what study designs are more suitable to give insights on 28 different types of questions. Decision-makers, evidence intermediaries (, researchers and funders can use this guidance to make better decisions on what type of study design to commission, use or fund when answering specific needs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mansilla et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Evidence networks facilitate the exchange of information and foster international relationships among researchers and stakeholders. These networks are instrumental in enabling the integration of scientific evidence into decision-making processes. While there is a global emphasis on evidence-based decision-making at policy and organisational levels, there exists a significant gap in our understanding of the most effective activities to exchange scientific knowledge and use it in practice. The objective of this rapid review was to explore the strategies employed by evidence networks to facilitate the translation of evidence into decision-making processes. This review makes a contribution to global health policymaking by mapping the landscape of knowledge translation in this context and identifying the evidence translation activities that evidence networks have found effective., Methods: The review was guided by standardised techniques for conducting rapid evidence reviews. Document searching was based on a phased approach, commencing with a comprehensive initial search strategy and progressively refining it with each subsequent search iterations. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) statement was followed., Results: The review identified 143 articles, after screening 1135 articles. Out of these, 35 articles were included in the review. The studies encompassed a diverse range of countries, with the majority originating from the United States (n = 14), followed by Canada (n = 5), Sweden (n = 2), and various other single locations (n = 14). These studies presented a varied set of implementation strategies such as research-related activities, the creation of teams/task forces/partnerships, meetings/consultations, mobilising/working with communities, influencing policy, activity evaluation, training, trust-building, and regular meetings, as well as community-academic-policymaker engagement., Conclusions: Evidence networks play a crucial role in developing, sharing, and implementing high-quality research for policy. These networks face challenges like coordinating diverse stakeholders, international collaboration, language barriers, research consistency, knowledge dissemination, capacity building, evaluation, and funding. To enhance their impact, sharing network efforts with wider audiences, including local, national, and international agencies, is essential for evidence-based decision-making to shape evidence-informed policies and programmes effectively., (© 2024. The Author(s).)

Rapid review methodology aims to facilitate faster conduct of systematic reviews to meet the needs of the decision-maker, while also maintaining quality and credibility. This systematic review aimed to determine the impact of different methodological shortcuts for undertaking rapid reviews on the risk of bias (RoB) of the results of the review. Review stages for which reviews and primary studies were sought included the preparation of a protocol, question formulation, inclusion criteria, searching, selection, data extraction, RoB assessment, synthesis, and reporting. We searched 11 electronic databases in April 2022, and conducted some supplementary searching. Reviewers worked in pairs to screen, select, extract data, and assess the RoB of included reviews and studies. We included 15 systematic reviews, 7 scoping reviews, and 65 primary studies. We found that several commonly used shortcuts in rapid reviews are likely to increase the RoB in the results. These include restrictions based on publication date, use of a single electronic database as a source of studies, and use of a single reviewer for screening titles and abstracts, selecting studies based on the full-text, and for extracting data. Authors of rapid reviews should be transparent in reporting their use of these shortcuts and acknowledge the possibility of them causing bias in the results. This review also highlights shortcuts that can save time without increasing the risk of bias. Further research is needed for both systematic and rapid reviews on faster methods for accurate data extraction and RoB assessment, and on development of more precise search strategies., (© 2023 Pan American Health Organization (PAHO/WHO); licensed by Research Synthesis Methods. Research Synthesis Methods published by John Wiley & Sons Ltd.)

Introduction: What are the different ways in which health equity can be sought through policy and programs? Although there is a central focus on health equity in global and public health, we recognize that stakeholders can understand health equity as taking different approaches and that there is not a single conceptual approach. However, information on conceptual categories of actions to improve health equity and/or reduce health inequity is scarce. Therefore, this study asks the research question: "what conceptual approaches exist in striving for health equity and/or reducing health inequity?" with the aim of presenting a comprehensive overview of approaches., Methods: A scoping review will be undertaken following the PRISMA guidelines for Scoping Reviews (PRISMA-ScR) and in consultation with a research librarian. Both the peer-reviewed and grey literatures will be searched using: Ovid MEDLINE, Scopus, PAIS Index (ProQuest), JSTOR, Canadian Public Documents Collection, the World Health Organization IRIS (Institutional Repository for Information Sharing), and supplemented by a Google Advanced Search. Screening will be conducted by two independent reviewers and data will be charted, coded, and narratively synthesized., Discussion: We anticipate developing a foundational document compiling categories of approaches and discussing the nuances inherent in each conceptualization to promote clarified and united action., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Amri et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Competing Interests: The authors have declared that no competing interests exist. Authors hold sole responsibility for the views expressed in the manuscript, which may not necessarily reflect the opinion or policy of the Pan American Health Organization.

Background: Evidence briefs for policy (EBP) draw on best-available data and research evidence (e.g., systematic reviews) to help clarify policy problems, frame options for addressing them, and identify implementation considerations for policymakers in a given context. An increasing number of governments, non-governmental organizations and research groups have been developing EBP on a wide variety of topics. However, the reporting characteristics of EBP vary across organizations due to a lack of internationally accepted standard reporting guidelines. This project aims to develop a STandard reporting guideline of Evidence briefs for Policy (STEP), which will encompass a reporting checklist and a STEP statement and a user manual., Methods: We will refer to and adapt the methods recommended by the EQUATOR (Enhancing the QUAlity and Transparency Of health Research) network. The key actions include: (1) developing a protocol; (2) establishing an international multidisciplinary STEP working group (consisting of a Coordination Team and a Delphi Panel); (3) generating an initial draft of the potential items for the STEP reporting checklist through a comprehensive review of EBP-related literature and documents; (4) conducting a modified Delphi process to select and refine the reporting checklist; (5) using the STEP to evaluate published policy briefs in different countries; (6) finalizing the checklist; (7) developing the STEP statement and the user manual (8) translating the STEP into different languages; and (9) testing the reliability through real world use., Discussion: Our protocol describes the development process for STEP. It will directly address what and how information should be reported in EBP and contribute to improving their quality. The decision-makers, researchers, journal editors, evaluators, and other stakeholders who support evidence-informed policymaking through the use of mechanisms like EBP will benefit from the STEP. Registration We registered the protocol on the EQUATOR network. ( https://www.equator-network.org/library/reporting-guidelines-under-development/#84 )., (© 2022. The Author(s).)

Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: TK, MK, LR, MR, JO, SiS, AR, MH, JR, AM, SoS are employed full-time with WHO, working in the fields of research, evidence and policy. SaS and CM had consultancy contracts with WHO, supporting the organization of the WHO Global Evidence-to-Policy Summit 2021. JL chaired the group of WHO Scientific Advisors of the WHO Global Evidence-to-Policy Summit and is co-lead of the Global Commission on Evidence to Address Societal Challenges. JB and EC were serving on the Executive Committee of the WHO Global Evidence-to-Policy Summit. The authors alone are responsible for the views expressed in this paper and they do not necessarily represent the views, decisions or policies of the institutions with which they are affiliated.

Introduction: The Coronavirus Disease 2019 (COVID19) pandemic has struck Latin America and the Caribbean (LAC) particularly hard. One of the crucial areas in the international community's response relates to accelerating research and knowledge sharing. The aim of this article is to map and characterise the existing empirical research related to COVID-19 in LAC countries and contribute to identify opportunities for strengthening future research., Methods: In this scoping review, articles published between December 2019 and 11 November 2020 were selected if they included an empirical component (explicit scientific methods to collect and analyse primary data), LAC population was researched, and the research was about the COVID-19 pandemic, regardless of publication status or language. MEDLINE, EMBASE, LILACS, Scielo, CENTRAL and Epistemonikos were searched. All titles and abstracts, and full texts were screened by two independent reviewers. Data from included studies was extracted by one reviewer and checked by a second independent reviewer., Results: 14,406 records were found. After removing duplicates, 5,458 titles and abstracts were screened, of which 2,323 full texts were revised to finally include 1,626 empirical studies. The largest portion of research came from people/population of Brazil (54.6%), Mexico (19.1%), Colombia (11.2%), Argentina (10.4%), Peru (10.3%) and Chile (10%), while Caribbean countries concentrated 15.3%. The methodologies most used were cross-sectional studies (34.7%), simulation models (17.5%) and randomized controlled trials (RCTs) (13.6%). Using a modified version of WHO's COVID-19 Coordinated Global Research Roadmap classification, 54.2% were epidemiological studies, followed by clinical management (22.3%) and candidate therapeutics (12.2%). Government and public funds support were reported in 19.2% of studies, followed by universities or research centres (9%), but 47.5% did not include any funding statement., Conclusion: During the first part of the COVID-19 pandemic, LAC countries have contributed to the global research effort primarily with epidemiological studies, with little participation on vaccines research, meaning that this type of knowledge would be imported from elsewhere. Research agendas could be further coordinated aiming to enhance shared self-sufficiency regarding knowledge needs in the region., Competing Interests: The authors have declared that no competing interests exist. Authors hold sole responsibility for the views expressed in the manuscript, which may not necessarily reflect the opinion or policy of the Pan American Health Organization nor the World Bank Group.

Background: Knowledge translation (KT) platforms are organisations, initiatives and networks that focus on supporting evidence-informed policy-making at least in part about the health-system arrangements that determine whether the right programmes, services and products get to those who need them. Many descriptions and evaluations of KT platforms in low- and middle-income countries have been produced but, to date, they have not been systematically reviewed., Methods: We identified potentially relevant studies through a search of five electronic databases and a variety of approaches to identify grey literature. We used four criteria to select eligible empirical studies. We extracted data about seven characteristics of included studies and about key findings. We used explicit criteria to assess study quality. In synthesising the findings, we gave greater attention to themes that emerged from multiple studies, higher-quality studies and different contexts., Results: Country was the most common jurisdictional focus of KT platforms, EVIPNet the most common name and high turnover among staff a common infrastructural feature. Evidence briefs and deliberative dialogues were the activities/outputs that were the most extensively studied and viewed as helpful, while rapid evidence services were the next most studied but only in a single jurisdiction. None of the summative evaluations used a pre-post design or a control group and, with the exception of the evaluations of the influence of briefs and dialogues on intentions to act, none of the evaluations achieved a high quality score., Conclusions: A large and growing volume of research evidence suggests that KT platforms offer promise in supporting evidence-informed policy-making in low- and middle-income countries. KT platforms should consider as next steps expanding their current, relatively limited portfolio of activities and outputs, building bridges to complementary groups, and planning for evaluations that examine 'what works' for 'what types of issues' in 'what types of contexts'.

The effects of housing instability and homelessness on child and adult health are well documented. However, few studies have explored health and housing interventions for families with children with the objective of health improvement. Housing Prescriptions as Health Care is a randomized controlled trial that is investigating the impact on physical and mental health of integrating priority placement in affordable housing and the provision of services (case management, financial, and legal), compared to the standard of care (providing resource guides and hospital-based social work or care navigation services). In 2016-19 seventy-eight homeless or housing-unstable families defined as "medically complex"-with a child or adult member who used more health services than usual or had a chronic disease or disability-were enrolled in the trial, and sixty-seven completed a six-month follow-up. A difference-in-differences analysis at six months showed decreases in the share of children in fair or poor health and in average anxiety and depression scores among parents in the intervention group, relative to the control group. Findings suggest that a population-specific model that integrates health, housing, legal, and social services can improve health-related outcomes at the household level.

In South America Andes hantavirus (ANDV) is hosted by the rodent Oligoryzomys longicaudatus (also known as pygmy rice rat). In humans, ANDV causes Hantavirus Pulmonary Syndrome (HPS), with a fatality rate of about 40%. Epidemiologic and molecular evidence has shown that ANDV can be transmitted from person to person. Sin Nombre hantavirus, occurring in North America, and ANDV are genetically related, and both cause HPS with similar clinical evolution and mortality rate. However, only ANDV is transmitted from person to person. A recent hantavirus outbreak in a small village in Southern Argentine, with 29 HPS cases and 11 deaths has brought to mind that person-to-person transmission continues to be a public health emergency. The present investigation was aimed to understand how does ANDV actually spread between persons. Tissue samples of lung and salivary glands from infected Oligoryzomys longicaudatus and lethal cases of human HPS were investigated by bright field immunocytochemistry, multichannel immunofluorescence, and transmission electron microscopy. The findings are consistent with ANDV infection and replication in the lung alveolar epithelium and macrophages, and in the secretory cells of the submandibular salivary glands. In the lung of infected Oligoryzomys longicaudatus and human cases HPS, the bulk of immunoreactive hantavirus antigens was localized in epithelial cells of the alveolar walls and macrophages. The ultrastructural study supports that in the lung of HPS patients the virus replicates in the alveolar epithelial cells with virus particles being discharged into the alveolar lumen. Virus-like particles were seen within vacuoles of the lung macrophages. Considering that these macrophages can reach the conductive segments of the airways, their expectoration becomes a deadly bullet for ANDV transmission. In the submandibular glands of infected rodents and HPS cases, ANDV antigens were in capillary endothelium, the secretory cells and filling the lumen of the excretory pathway. It is proposed that in patients with HPS caused by ANDV the alveolar epithelium and macrophages would be the gate for the airway spreading of the virus, while the salivary glands are a target for virus replication and an exit pathway through saliva., (Copyright © 2020 Pizarro, Navarrete, Mendez, Zaror, Mansilla, Tapia, Carrasco, Salazar, Murua, Padula, Otth and Rodríguez.)

Background: Affordability is a key barrier to access to medicines. Generic medicines policies can address this barrier and promote access. Successful uptake of generic medicines depends, in part, on ensuring that these products are interchangeable with reference products. Typically, bioequivalence certification is established in order to demonstrate such interchangeability., Objective: To study the implementation of the bioequivalence certification policy in Chile., Methods: We used Chilean Market Regulatory Authority data for analysis to study the number of products that obtained bioequivalence certification, the time until bioequivalence certification and associated factors to obtain bioequivalence., Results: As of January 2017, out of 2,336 products with a valid market authorization containing at least one of the 167 APIs that required BE certification, 1,026 products actually have BE certification (1,026/2,336, 43.9% compliance). Where data were available, the time between submission of the market authorization as a bioequivalent product to final authorization by the national medicine regulatory authority for most products varied between 4-6 months. The fraction of all BE products containing a given API out of the total marketed products containing that API varies considerably, e.g. for the API olmesartan there was only a single BE product marketed, the API diclofenac had none., Conclusions: Although the implementation of Chile's bioequivalence policy increased the number of bioequivalent products, over 50% of generic products requiring bioequivalence that did not obtain this certification. Also for some of the API none or very few BE products are marketed which limits the success of a substitution policy. Further studies are required to identify the apparent lack of incentives to obtain bioequivalence certification. Studies of sales volumes and prices of the products are needed to identify whether generic products without bioequivalence certification either become bioequivalent or eventually exit the market., Competing Interests: The authors have declared that no competing interests exist.

The potential human health risks following the exposure to inorganic nanoparticles (NPs) is a very important issue for their application in leather finishing industry. The aim of our study was to investigate the cytotoxic effect of silver (Ag)/titanium dioxide (TiO 2 ) NPs on human cells. Photocatalytic NPs were prepared by electrochemical deposition of Ag on the surface of TiO 2 and nitrogen (N)-TiO 2 NPs and, subsequently, physico-chemical characterized. Then, a set of experiments have been performed to study the cytotoxicity and cell death mechanisms involved, the changes in cell morphology and the production of ROS induced in human keratinocytes (HaCaT) and human lung epithelial cells (A549) by exposure to NPs. Moreover, the changes in major signaling pathways and the inflammatory response induced by Ag/N-TiO 2 NPs in A549 cells were investigated. The data showed that cell death by late apoptosis/necrosis is induced in cells as function of the dose and the type of NPs and is characterized by morphological changes and cytoskeletal disorganization and an increase in reactive oxygen species (ROS) production. The exposure of A549 cells to Ag/N-TiO 2 NPs determine the activation of ERK1/2 MAP-kinase pathway and the release of pro-inflammatory mediators CXCL1, GM-CSF and MIF, known to be involved in the recruitment of circulating neutrophils and monocytes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Introduction: Chile implemented a generic substitution policy in 2014 to improve access to medicines. This study aims to measure if the generic substitution policy had an effect on the sales volume and prices of referent and the branded generic products with demonstrated bioequivalence (BEQ) in the private pharmaceutical market., Methods: The volume and total private sales of medicines sold at private sector retail outlets between November 2011 and October 2016 were considered in the analysis. We calculated the total number of daily defined doses (DDD) by adding up the number of DDDs of different presentations with the active pharmaceutical ingredient (API). We determined the ratio of the median prices of all BEQ per DDD presentations compared with the median price of the corresponding referent presentations per DDD in 2011 and 2016. Sixteen APIs representing 231 different conventional-release presentations were included in the analysis., Results: Overall, the volume of sales of the referent products decreased over time after the intervention. However, this reduction was not mirrored by an increase in the corresponding branded generic BEQ volumes overall. In all cases, the median price per DDD of the referent was higher than its BEQ counterpart in 2011 and 2016., Conclusion: Since referent products are more costly than branded BEQ generic products, reducing their consumption-and increasing the BEQ availability-should improve access to medicines in Chile. However, this must be accompanied by promotion of BEQ products to ensure savings for consumers in the long term. Future research should focus on identifying facilitating and inhibiting factors of generic substitution.

Introduction: The ROP Argentina Group was created in 2003 and is responsible for the National Program for the Prevention of Blindness in Childhood by Retinopathy of Prematurity (ROP) in Argentina., Objetives: To describe the program implementation and results achieved in relation to ROP care in terms of effectiveness, access, and quality (2004-2016)., Population and Methods: Descriptive, retrospective study with a dynamic cohort carried out in facilities that are part of the registry. Eligible population: All preterm newborn infants with risk factors for ROP., Results: Participating health care services increased from 14 to 98 and covered the 23 provinces and the Autonomous City of Buenos Aires. A total of 956 infants were born with < 1500 g in 2004 and 2739, in 2016. Of these, 22.7 % had some degree of ROP and 7.8 % required treatment (severe ROP). Vision screening exceeded 90 %, and treatments at the place of origin increased (57 %-92 %). The incidence of unusual cases is still high (17.3 % of treated cases), and missed opportunities are still recorded. The use of anti-angiogenic drugs trebled since 2011, when they started to be used., Conclusions: Significant achievements were observed in terms of program representativeness, scope, and adherence, and also in relation to screening access and treatment at the place of origin; however, the incidence of ROP is still high. The persistence of unusual cases and missed opportunities evidences deficiencies in the quality of health care and outpatient followup and underlines the need to strengthen the program actions in relation to services., (Sociedad Argentina de Pediatría.)

We present a complete analysis of the photoinjection process in 3 nm spherical ZnO nanoparticles with different dyes attached to their surface. The study was carried using a TD-DFTB model to describe the quantum dynamics of charge transfer in the presence of an external time dependent electric field simulating resonant illumination. The ZnO + dye systems have been classified according to the injection process (type I and type II) that they present. In addition, the calculated charge transfer shows that many dyes present hole injection into the nanoparticle. We have classified the charge transfer into holes or electron and compare these results with the TiO2 + dye system obtained from the literature.

Multiple myeloma (MM) is a very heterogeneous disease, characterized by multiple cytogenetic aberrations on plasma cells (PC) that have been traditionally used to predict the outcome of the disease. A mayor issue on the analysis of PC is the sometimes low infiltration of these cells in the bone marrow that hampers cytogenetic studies. To solve this problem we have optimized a selection strategy based on PC immunomagnetic isolation that has allowed us to lower to 1% the minimal PC infiltration requirement without loss of purity, enabling to perform genetic analysis. In this study, we have analyzed 153 bone marrow samples of patients suspected of MM, collected from February 2015 to May 2017 by the Genetics service of the Complejo Hospitalario de Navarra. Clinical characteristics of the patients and PC immunophenotyping, conventional cytogenetics and interphase fluorescence in situ hybridization (iFISH) analyses have been assessed on these samples. In our cohort 90% of the samples had cytogenetic abnormalities, among them 50% presented immunoglobulin rearrangements, 41.9% showed 1q gains, 29.7% showed 1p deletions and 33% presented TP53 deletion., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

Liquid biopsy represents an alternative to conventional biopsies for the evaluation of tumors mainly due to its easy sampling. One of the main applications is the enumeration of Circulating Tumor Cells (CTCs) to evaluate tumor progression or response to treatment. The analysis of the functional characteristics of CTCs could give us much more information about their role in order to establish a more personalized treatment for the patients. The major issue that has to be solved is the isolation of the CTC population. Multiple protocols have been developed, however none of them has demonstrated to be the definitive one. In fact, a combination of these techniques has often been performed in order to obtain a purer and viable population of CTCs. In this review we have summarized for the first time the different combinatorial approaches used in the last years to optimize the isolation of CTCs and their limitations., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Epstein-Barr (EBV) and Adenovirus (AdV) viral infections represent a significant cause of morbi-mortality in allogeneic hematopoietic stem cell transplantation recipients due to the use of immunomyelosuppressive treatments and the prolonged period of immunodeficiency that they generate. To date, the post-transplant protective role of EBV and AdV specific CD8+ T lymphocytes (CTLs) has been demonstrated. However, other factors are increasingly important in regulating the reconstitution and activity of CTLs specific to these viruses such as different cell subpopulations (CD4 + T lymphocytes, regulatory T lymphocytes, dendritic cells, Natural Killer cells, etc.), molecular mechanisms of immunoregulation and the drugs administered to the patient as prophylaxis for a possible graft-versus-host disease. The aim of this review is to analyze the importance of monitoring the functional EBV and AdV-specific cellular response in the management of post-transplant recipients.

Overweight and obesity are a global epidemic with rates having risen to alarming levels in both developed and developing countries. Chile has been no exemption, with sharp increases in obesity prevalence, especially among school-age children. This paper describes the policy actions and strategies implemented to tackle this major public health concern in Chile over the last 10 years, and highlights the main challenges and nuances of the process. Chile has taken policy action that includes front-of-package labelling, advertising regulations, and school-food restrictions. New policies focus on the social determinants of health as they relate to food environments and people's behavior. These actions are not only suitable to the current context in Chile, but are also supported by the best available scientific evidence. Moreover, the implementation of these policies has produced a broad debate involving public institutions and the food industry, with discussions issues ranging from property rights to trade barriers. Despite some differences among stakeholders, a valuable political consensus has been achieved, and several international organizations are eager to evaluate the impact of these pioneer initiatives in Latin America ., Competing Interests: Conflict of interests: None declared.

The use of preemptive antiviral therapy to prevent cytomegalovirus (CMV) disease in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients might result in over-treatment, inducing drug-related toxicity and viral resistance. A search for predictive markers is needed to determine requirement for antiviral therapy. Clinical follow-up, in combination with the use of streptamers (STs) and cytokine-intracellular staining, could help to identify patients at high risk for CMV reactivations. To study the immune response and reactivation control by CMV-specific CD8 + T-cell (CMV-CTL) populations, we monitored 25 patients who have undergone allo-HSCT by using ST multimer and intracellular cytokine staining. Our study has revealed that the presence of functional CMV-specific T cells, determined by early interferon γ production or by significant T-cell expansion after first CMV reactivation, correlated with short CMV viremia duration and low number of CMV reactivations. By contrast, the absence of functional CMV-CTLs does correlate with CMV recurrence. These results support that behavior of CMV-specific subpopulations after reactivation influences reactivations and can guide preemptive therapy., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Informing the health policymaking process with the best available scientific evidence has become relevant to health systems globally. Knowledge Translation Platforms (KTP), such as the World Health Organization's Evidence Informed Policy Networks (EVIPNet), are a recognized strategy for linking research to action. This report describes the experience of implementing EVIPNet in Chile, from its objectives, organizational structure, strategy, activities, and main outputs, to its evolution over the course of its first year. Lessons learned are also covered. Of the activities initiated by EVIPNet-Chile, the Rapid Response Service proved to be a good starting point for engaging policymakers. Capacity building workshops and policy dialogues with relevant stakeholders were also successful. Additionally, EVIPNet-Chile developed a model for engaging academic institutions in policymaking through a network focused on preparing evidence briefs. A number of challenges, such as changing methods for producing rapid evidence syntheses, were also identified. This KTP implementation model located in a Ministry of Health could contribute to the development of similar initiatives in other health systems., Competing Interests: Conflict of interests. None declared.

Background: Multimer technology is widely used to screen antigen-specific immune recovery after allogeneic hematopoietic stem cell transplantation (allo-HSCT) as it enables identification, enumeration, phenotypic characterization and isolation of virus-specific T-cells. Novel approaches of multimerization might improve on classical tetramer staining; however, their use as standard monitoring technique to quantify antigen-specific cells has not been validated yet. We have compared two of these available multimeric complexes: pentamer and streptamer to select the best strategy for the incorporation into clinical monitoring practice., Methods: CMVpp65 495-503 -specific HLA-A*02:01 CD8 + T lymphocytes (CTL A * 02:01 -CMVpp65 495-503 ) were examined with pentamer and streptamer in peripheral blood cells of 77 healthy volunteers. Quantitative and qualitative analyses were performed to compare the precision and repeatability, sensitivity and accuracy and specificity of both technologies by flow cytometry., Results: Standard deviation for both techniques was less than 0.05 showing that they are repetitive and precise. Both techniques significantly correlated at high frequencies (r Spearman  = 0.9422; P < 0.0001) but it was lost at lower levels (<1%) of CTL A * 02:01 -CMVpp65 495-503 (r Spearman  = 0.3351; P = 0.1376). Streptamer is more accurate for the detection of CTL A * 02:01 -CMVpp65 495-503 providing significantly closer values to the theoretical ones (P < 0.0001) as pentamer binds unspecifically to a notable proportion of non-CMV-specific CD8 + T-cells., Conclusion: Our results suggest that streptamer multimer provides precise, accurate and specific results to detect CTL A * 02:01 -CMVpp65 495-503 by flow cytometry. Streptamer multimer can be used not only for the monitoring of early CTL A * 02:01 -CMVpp65 495-503 reconstitution in immunosuppressed patients following allo-HSCT but also, in conjunction with its reversibility role, for the isolation of CTL A * 02:01 -CMVpp65 495-503 for its future use in adoptive immunotherapy. © 2016 International Clinical Cytometry Society., (© 2016 International Clinical Cytometry Society.)

In recent years there has been significant debate on whether the edge type of graphene nanoflakes (GNFs) or graphene quantum dots (GQDs) are relevant for their electronic structure, thermal stability, and optical properties. Using computer simulations, we have proven that there is a fundamental difference in the absorption spectra between samples of the same shape, similar size but different edge type, namely, armchair or zigzag edges. These can be explained by the presence of electronic structures near the Fermi level which are localized on the edges. These features are also evident from the dependence of band gap on the GNF size, which shows three very distinct trends for different shapes and edge geometries.

Elongated flexuous plant viral nanoparticles (VNPs) represent an interesting platform for developing different applications in nanobiotechnology. In the case of potyviruses, the virion external surface is made up of helically arrayed domains of the viral structural coat protein (CP), repeated over 2000 times, in which the N- and C-terminal domains of each CP are projected toward the exterior of the external virion surface. These characteristics provide a chemical environment rich in functional groups susceptible to chemical conjugations. We have conjugated Candida antarctica lipase B (CALB) onto amino groups of the external surface of the potyvirus turnip mosaic virus (TuMV) using glutaraldehyde as a conjugating agent. Using this approach, TuMV virions were transformed into scaffolds for CALB nanoimmobilization. Analysis of the resulting structures revealed the formation of TuMV nanonets onto which large CALB aggregates were deposited. The functional enzymatic characterization of the CALB-bearing TuMV nanonets showed that CALB continued to be active in the nanoimmobilized form, even gaining an increased relative specific activity, as compared to the non-immobilized form. These novel virus-based nanostructures may provide a useful new approach to enzyme nanoimmobilization susceptible to be industrially exploited.

Background: It is of utmost importance to identify hypertensive subjects in a country, in order to use efficiently public resources. The National Health Survey 2009-10 in Chile incorporated a third measurement of blood pressure (BP) during the home visit performed by a nurse, and a second day of measurement in a representative sub-sample., Aim: To study the effect of these two additional actions over both the average value of BP and the national prevalence of hypertension., Material and Methods: A third blood pressure measurement was carried out in 5,058 subjects, and it was measured in a second day in 930 individuals. The effect of these additional measurements on absolute blood pressure values and the prevalence of hypertension were assessed., Results: A small but statistically significant reduction in mean systolic pressure (0.52 mmHg) and the prevalence of hypertension (1%) was observed after the incorporation of the third blood pressure measurement. No effects in these figures were observed after the measurement performed on a second day., Conclusions: These findings should be considered when designing the new National Health Survey in Chile.

Peripheral blood progenitor cells (PBPCs) have become the major source of hematopoietic progenitor cells for allogeneic transplantation. In February 2008, Zarzio® was approved by the European Medicine Agency for PBPCs mobilization, but this authorization was not based in trials analyzing safety and efficacy for PBPCs mobilization. Since August 2011, Zarzio® has been used at our institution for PBPCs mobilization. In total 36 healthy family donors underwent PBPCs mobilization, 18 with Neupogen® and 18 with Zarzio®. Donor characteristics were equivalent between groups, and no severe adverse effects were registered in the Zarzio® group. The number of CD34 cells collected/Kg recipient body weight was 6.7 × 10(6) (3.8-11.1) in the Zarzio® group versus 8.4 × 10(6) (5.6-16.6) in the Neupogen® group (P = 0.04). We collected the minimal target cell dose (2 × 10(6) /kg) in all donors from each group and no significant differences were found in the collection of the optimal cell dose (5 × 10(6) /kg) between groups, although 3/18 (16.6%) donors that received Zarzio® failed to mobilize the optimal cell dose compared with 0% in the Neupogen® group. A total of 35 patients proceeded to transplantation (17 in the Zarzio® and 18 in the Neupogen® groups, respectively). Platelet and neutrophil median time to engraftment was comparable between the two groups. Our retrospective study supports the conclusion that Zarzio® mobilization of PBPCs in healthy donors is safe but perhaps not as effective as the reference Neupogen. However, more prospective trials are required to definitively asses the safety and efficacy of G-CSF biosimilars for PBPCs mobilization in healthy donors., (© 2015 Wiley Periodicals, Inc.)

Turnip mosaic virus (TuMV) infections affect many Arabidopsis developmental traits. This paper analyzes, at different levels, the development-related differential alterations induced by different strains of TuMV, represented by isolates UK 1 and JPN 1. The genomic sequence of JPN 1 TuMV isolate revealed highest divergence in the P1 and P3 viral cistrons, upon comparison with the UK 1 sequence. Infectious viral chimeras covering the whole viral genome uncovered the P3 cistron as a major viral determinant of development alterations, excluding the involvement of the PIPO open reading frame. However, constitutive transgenic expression of P3 in Arabidopsis did not induce developmental alterations nor modulate the strong effects induced by the transgenic RNA silencing suppressor HC-Pro from either strain. This highlights the importance of studying viral determinants within the context of actual viral infections. Transcriptomic and interactomic analyses at different stages of plant development revealed large differences in the number of genes affected by the different infections at medium infection times but no significant differences at very early times. Biological functions affected by UK 1 (the most severe strain) included mainly stress response and transport. Most cellular components affected cell-wall transport or metabolism. Hubs in the interactome were affected upon infection.

Approximately, up to 70 % of the human population is infected with cytomegalovirus (CMV) that persists for life in a latent state. In healthy people, CMV reactivation induces the expansion of CMV-specific T cells up to 10 % of the entire T cell repertoire. On the contrary, CMV infection is a major opportunistic viral pathogen that remains a leading cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation. Due to the delayed CMV-specific immune recovery, the incidence of CMV reactivation during post-transplant period is very high. Several methods are currently available for the monitoring of CMV-specific responses that help in clinical monitoring. In this review, essential aspects in the immune recovery against CMV are discussed to improve the better understanding of the immune system relying on CMV infection and, thereby, helping the avoidance of CMV disease or reactivation following hematopoietic stem cell transplantation with severe consequences for the transplanted patients.

We performed a prospective phase II study to evaluate clinical safety and outcome in 48 patients with steroid-refractory grade II-IV acute graft-versus-host disease (aGVHD) treated with mesenchymal stromal cells (MSCs). Clinical outcomes were correlated to comprehensive analyses of soluble and cellular biomarkers. Complete resolution (CR) of aGVHD at day 28 (CR-28) occurred in 12 (25%) patients, CR lasting >1 month (CR-B) occurred in 24 (50%) patients. One-year overall survival was significantly improved in CR-28 (75 versus 33%, P=0.020) and CR-B (79 versus 8%, P<0.001) versus non-CR patients. A six soluble biomarker-panel was predictive for mortality (HR 2.924; CI 1.485-5.758) when measured before MSC-administration. Suppression of tumorigenicity 2 (ST2) was only predictive for mortality 2 weeks after but not before MSC-administration (HR 2.389; CI 1.144-4.989). In addition, an increase in immature myeloid dendritic cells associated with decreased mortality (HR 0.554, CI 0.389-0.790). Patients had persisting T-cell responses against defined virus- and leukemia-associated antigens. In conclusion, our data emphasize the need to carefully assess biomarkers in cohorts with homogeneous GVHD treatments. Biomarkers might become an additional valuable component of composite end points for the rapid and efficient testing of novel compounds to decrease lifecycle of clinical testing and improve the success rate of phase II/III trials.

Background: Adoptive transfer of CMV-specific T cells has shown promising results in preventing pathological effects caused by opportunistic CMV infection in immunocompromised patients following allogeneic hematopoietic stem cell transplantation. The majority of studies have used steady-state leukapheresis for CMV-reactive product manufacture, a collection obtained prior to or months after G-CSF mobilization, but the procurement of this additional sample is often not available in the unrelated donor setting. If the cellular product for adoptive immunotherapy could be generated from the same G-CSF mobilized collection, the problems associated with the additional harvest could be overcome. Despite the tolerogenic effects associated with G-CSF mobilization, recent studies described that CMV-primed T cells generated from mobilized donors remain functional., Methods: MHC-multimers are potent tools that allow the rapid production of antigen-specific CTLs. Therefore, in the present study we have assessed the feasibility and efficacy of CMV-specific CTL manufacture from G-CSF mobilized apheresis using MHC-multimers., Results: CMV-specific CTLs can be efficiently isolated from G-CSF mobilized samples with Streptamers and are able to express activation markers and produce cytokines in response to antigenic stimulation. However, this anti-viral functionality is moderately reduced when compared to non-mobilized products., Conclusions: The translation of Streptamer technology for the isolation of anti-viral CTLs from G-CSF mobilized PBMCs into clinical practice would widen the number of patients that could benefit from this therapeutic strategy, although our results need to be taken into consideration before the infusion of antigen-specific T cells obtained from G-CSF mobilized samples.

This work presents a new approach to obtain reliable surface topography reconstructions from 2D Scanning Electron Microscopy (SEM) images. In this method a set of images taken at different tilt angles are compared by means of digital image correlation (DIC). It is argued that the strength of the method lies in the fact that precise knowledge about the nature of the rotation (vector and/or magnitude) is not needed. Therefore, the great advantage is that complex calibrations of the measuring equipment are avoided. The paper presents the necessary equations involved in the methods, including derivations and solutions. The method is illustrated with examples of 3D reconstructions followed by a discussion on the relevant experimental parameters., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: Cytomegalovirus (CMV)-specific T cell infusion to immunocompromised patients following allogeneic Hematopoietic Stem Cell Transplantation (allo-HSCT) is able to induce a successful anti-viral response. These cells have classically been manufactured from steady-state apheresis samples collected from the donor in an additional harvest prior to G-CSF mobilization, treatment that induces hematopoietic stem cell (HSC) mobilization to the periphery. However, two closely-timed cellular collections are not usually available in the unrelated donor setting, which limits the accessibility of anti-viral cells for adoptive immunotherapy. CMV-specific cytotoxic T cell (CTL) manufacture from the same G-CSF mobilized donor stem cell harvest offers great regulatory advantages, but the isolation using MHC-multimers is hampered by the high non-specific binding to myeloid progenitors, which reduces the purity of the cellular product., Methods: In the present study we describe an easy and fast method based on plastic adherence to remove myeloid cell subsets from 11 G-CSF mobilized donor samples. CMV-specific CTLs were isolated from the non-adherent fraction using pentamers and purity and yield of the process were compared to products obtained from unmanipulated samples., Results: After the elimination of unwanted cell subtypes, non-specific binding of pentamers was notably reduced. Accordingly, following the isolation process the purity of the obtained cellular product was significantly improved., Conclusions: G-CSF mobilized leukapheresis samples can successfully be used to isolate antigen-specific T cells with MHC-multimers to be adoptively transferred following allo-HSCT, widening the accessibility of this therapy in the unrelated donor setting. The combination of the clinically translatable plastic adherence process to the antigen-specific cell isolation using MHC-multimers improves the quality of the therapeutic cellular product, thereby reducing the clinical negative effects associated with undesired alloreactive cell infusion.

Chemotherapy and/or radiotherapy regular regimens used for conditioning of recipients of hematopoietic stem cell transplantation (SCT) induce a period of transient profound immunosuppression. The onset of a competent immunological response, such as the appearance of viral-specific T cells, is associated with a lower incidence of viral infections after haematopoietic transplantation. The rapid development of immunodominant peptide virus screening together with advances in the design of genetic and non-genetic viral- and tumoural-specific cellular selection strategies have opened new strategies for cellular immunotherapy in oncologic recipients who are highly sensitive to viral infections. However, the rapid development of cellular immunotherapy in SCT has disclosed the role of the T cell selection method in the modulation of functional cell activity and of in vivo secondary effects triggered following immunotherapy.

Losses produced by virus diseases depend mostly on symptom severity. Turnip mosaic virus (TuMV) is one of the most damaging and widespread potyvirus infecting members of the family Brassicaceae, including Arabidopsis thaliana. We used JPN1 and UK1 TuMV strains to characterize viral infections regarding symptom development, senescence progression, antioxidant response, reactive oxygen species (ROS) accumulation, and transcriptional profiling. Both isolates, despite accumulating similar viral titers, induced different symptomatology and strong differences in oxidative status. Early differences in several senescence-associated genes linked to the ORE1 and ORS1 regulatory networks as well as persistent divergence in key ROS production and scavenging systems of the plant were detected. However, at a later stage, both strains induced nutrient competition, indicating that senescence rates are influenced by different mechanisms upon viral infections. Analyses of ORE1 and ORS1 levels in infected Brassica juncea plants showed a similar pattern, suggesting a conserved differential response to both strains in Brassicaceae spp. Transcriptional analysis of the ORE1 and ORS1 regulons showed similarities between salicylic acid (SA) response and the early induction triggered by UK1, the most severe strain. By means of SA-defective NahG transgenic plants, we found that differential senescence progression and ROS accumulation between strains rely on an intact SA pathway.

Multimers are complexes of recombinant MHC-class I molecules conjugated with antigenic immunodominant peptides and labeled with fluorescent molecules or magnetic microbeads that allow the quantification and selection of virus-specific cytotoxic T-cell subpopulations. Specific T-cell receptors recognize the immunodominant peptides and bind to the multimers. Although these complexes are only recognized by CD8(+) T cells with specific T-cell receptors for the particular antigen, it has been observed that multimers can also bind non-specifically to CD8- cells, such as B-cells and monocytes. Using PBMCs from CMV-seropositive healthy donors, we analyze the tendency of Pentamer and Streptamer multimers towards non-specific interactions and describe a method to avoid this unwanted event. We find that a notable proportion of multimer-positive cells are likely to represent cross-contamination by cells lacking a TCR specific for pp65. In addition, we demonstrate that this unspecific interaction can be overcome by the pre-incubation of multimer-stained PBMCs with human AB serum, without altering their capacity to bind specifically to the CD8(+) T cell population of interest. In conclusion, in this study we characterize a novel method to abrogate TCR-independent interactions of multimers to ensure a pure and safe therapeutic product for Adoptive Immunotherapy., (© 2013. Published by Elsevier B.V. All rights reserved.)

Orbital cellulitis typically occurs in older children, but it can occasionally affect infants and neonates. Staphylococcus aureus is the main pathogen isolated. Outcome depends on an adequate initial approach. We report three neonates with orbital cellulitis caused by community-associated MRSA.

Cervical carcinoma is one of the most common cancers in women worldwide. It is well established that chronic infection of the genital tract by various mucosatropic human papillomavirus (HPV) types causes cervical cancer. Cellular immunity to E7 protein from HPV (HPVE7) has been associated with clinical and cytologic resolution of HPV-induced lesions. Thus, we decided to test if targeting of HPVE7 to dendritic cells using a fusion protein containing the extra domain A (EDA) from fibronectin, a natural ligand for TLR4, and HPVE7 (EDA-HPVE7) might be an efficient vaccine for the treatment of cervical carcinoma. We found that EDA-HPVE7 fusion protein was efficiently captured by bone marrow derived dendritic cells in vitro and induced their maturation, with the upregulation of maturation markers and the production of IL-12. Immunization of mice with EDA-HPVE7 fusion protein induced antitumor CD8(+) T cell responses in the absence of additional adjuvants. Repeated intratumoral administration of EDA-HPVE7 in saline was able to cure established TC-1 tumors of 5-7 mm in diameter. More importantly, intravenous injection with EDA-HPVE7 in combination with the TLR ligand polyinosinic-polycytidylic acid (pIC), or with low doses of cyclophosphamide and the TLR9 ligand CpG-B complexed in cationic lipids, were able to eradicate large established TC-1 tumors (1.2 cm in diameter). Thus, therapeutic vaccination with EDA-HPVE7 fusion protein may be effective in the treatment of human cervical carcinoma., (Copyright © 2011 UICC.)

The Extradomain A from fibronectin (EDA) has an immunomodulatory role as fusion protein with viral and tumor antigens, but its effect when administered with bacteria has not been assessed. Here, we investigated the adjuvant effect of EDA in mice immunizations against Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella Enteritidis). Since lipopolysaccharide (LPS) is a major virulence factor and the LPS O-polysaccharide (O-PS) is the immunodominant antigen in serological diagnostic tests, Salmonella mutants lacking O-PS (rough mutants) represent an interesting approach for developing new vaccines and diagnostic tests to differentiate infected and vaccinated animals (DIVA tests). Here, antigenic preparations (hot-saline extracts and formalin-inactivated bacterins) from two Salmonella Enteritidis rough mutants, carrying either intact (SEΔwaaL) or deep-defective (SEΔgal) LPS-Core, were used in combination with EDA. Biotinylated bacterins, in particular SEΔwaaL bacterin, decorated with EDAvidin (EDA and streptavidin fusion protein) improved the protection conferred by hot-saline or bacterins alone and prevented significantly the virulent infection at least to the levels of live attenuated rough mutants. These findings demonstrate the adjuvant effect of EDAvidin when administered with biotinylated bacterins from Salmonella Enteritidis lacking O-PS and the usefulness of BEDA-SEΔwaaL as non-live vaccine in the mouse model.

The development of effective vaccines against HIV-1 infection constitutes one of the major challenges in viral immunology. One of the protein candidates in vaccination against this virus is p24, since it is a conserved HIV antigen that has cytotoxic and helper T cell epitopes as well as B cell epitopes that may jointly confer enhanced protection against infection when used in immunization-challenge approaches. In this context, the adjuvant effect of EDA (used as EDAp24 fusion protein) and poly(I:C), as agonists of TLR4 and TLR3, respectively, was assessed in p24 immunizations using a recombinant Listeria monocytogenes HIV-1 Gag proteins (Lm-Gag, where p24 is the major antigen) for challenge in mice. Immunization with EDAp24 fusion protein together with poly(I:C) adjuvant induced a specific p24 IFN-γ production (Th1 profile) as well as protection against a Lm-Gag challenge, suggesting an additive or synergistic effect between both adjuvants. The combination of EDA (as a fusion protein with the antigen, which may favor antigen targeting to dendritic cells through TLR4) and poly(I:C) could thus be a good adjuvant candidate to enhance the immune response against HIV-1 proteins and its use may open new ways in vaccine investigations on this virus., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Orbital cellulitis typically occurs in older children, but it can occasionally affect infants and neonates. Staphylococcus aureus is the main pathogen isolated. Outcome depends on an adequate initial approach. We report three neonates with orbital cellulitis caused by community-associated MRSA.

Adjuvant research is being oriented to TLR-agonists, but complement activation has been relatively unexplored. In previous studies it was demonstrated that poly(methyl vinyl ether-co-maleic anhydride) nanoparticles (PVMA NPs) used as adjuvant differentially activate dendritic cells through toll like receptors (TLR) stimulation, however, a high dose of these NPs was used. Now, we demonstrated a dose-response effect, with a concentration as low as 20μg/mL able to stimulate TLR2 and TLR4 transfected dendritic cells. In addition, we investigated whether PVMA NPs are able to exploit also the immunomodulatory benefits of complement activation. Results indicated that the hydroxylated surface of these NPs highly activated the complement cascade, as measured by adsorption studies and a complement fixation bioassay. Stable binding of C3b to NPs was confirmed as indicated by lability to SDS treatment after washing resistance. Complement consumption was confirmed as the lytic capacity of complement exposed to NPs was abolished against antibody-sensitized sheep erythrocytes, with a minimal inhibitory concentration of 50μg NPs, equivalent to a surface of 1cm(2). On the contrary, nanoparticles prepared with poly(lactic-co-glycolic acid) (PLGA), used as a reference, did not consume complement at a concentration ≥3mg NPs (≥40cm(2)). Complement consumption was inhibited when PVMA NPs were cross-linked with diamino groups (1,3-diaminopropane), indicating the role of hydroxyl groups as responsible of the phenomenon. These results favour a model whereby PVMA NPs adjuvant activate complement on site to attract immature antigen presenting cells that are activated through TLR2 and TLR4., (Copyright © 2011 Elsevier Ltd. All rights reserved.)