Long-term care for older people is provided in both residential and non-residential settings, with residential settings tending to cater for individuals with higher care needs. Evidence relating to the costs and effectiveness of different workforce structures and care processes is important to facilitate the future planning of residential aged care services to promote high quality care and to enhance the quality of life of individuals living in residential care. A systematic review conducted up to December 2015 identified 19 studies containing an economic component; seven included a complete economic evaluation and 12 contained a cost analysis only. Key findings include the potential to create cost savings from a societal perspective through enhanced staffing levels and quality improvement interventions within residential aged care facilities, while integrated care models, including the integration of health disciplines and the integration between residents and care staff, were shown to have limited cost-saving potential. Six of the 19 identified studies examined dementia-specific structures and processes, in which person-centred interventions demonstrated the potential to reduce agitation and improve residents’ quality of life. Importantly, this review highlights methodological limitations in the existing evidence and an urgent need for future research to identify appropriate and meaningful outcome measures that can be used at a service planning level. The United Nations has reported population ageing in nearly every country in the world and projections suggest that the number of people aged 60 and over will more than double the 2013 level by 2050 [Aged care is a significant responsibility for governments. In most OECD countries, aged care accounts for roughly 1–1.5% of GDP in terms of government funding , and on Residential care is in the midst of a ‘culture change’ movement, involving organisational change and a move toward providing more person-centred, individualised care . Person-The framework of economic evaluation is increasingly being applied in health and aged care services in an effort to promote efficiency in the design and delivery of services. Knowledge of the incremental costs and effectiveness of differing program design features is essential for well-informed resource allocation decisions in residential care. Program design features can be broken down into subcategories to assist in the assessment of quality (see Donabedian ). This rWhich structures and processes in residential aged care settings have been demonstrated to be cost effective?How have the costs and outcomes for residents with dementia been assessed in economic evaluations?To this end, the main objectives of this review were to answer the following questions:http://www.crd.york.ac.uk/PROSPERO; registration number CRD42015015977).A protocol for this systematic review was registered with the PROSPERO International Prospective Register of Systematic Reviews on 30 January 2015 , partial economic evaluations , and randomised trials reporting more limited information, such as estimates of resource use or costs of interventions, pertaining to structures and processes of care aimed at improving the quality of care for older adults in a residential aged care setting.Structures of care were defined as the workforce and its operations, and included level of staffing, expertise of staff, hours of care per resident per day, and continuity of care. Processes of care included activity programs and services implemented in the context of care provision. These definitions were adapted from Donabedian’s quality of care model incorporating structure, process, and outcome [ outcome .Studies pertaining to interventions that did not apply at a facility or unit level such as individualised pharmaceutical interventions and feeding tubes were excluded from this review.Eight electronic bibliographic databases were searched from inception to the 8th October 2014, including AgeLine, CINAHL, Econlit, Informit , Medline, ProQuest, Scopus, and Web of Science. An update search was run on 14 December 2015.The search strategies were developed and reviewed with the assistance of two Health Sciences Librarians with expertise in systematic reviews. The strategy combined terms relating to nursing homes, economics, and older people, limited to English language. No study design or date limits were imposed on the search. The full search strategy is available on PROSPERO.Due to the large number of results retrieved when searching the multidisciplinary database ProQuest, results were limited to scholarly journals, reports, dissertations and theses, conference papers and proceedings, and working papers. Newspapers, trade journals, wire feeds, magazines, other sources, books, and encyclopedias and reference works were excluded.Titles and abstracts of studies retrieved were reviewed in full by the primary review author. A second reviewer independently screened 10% of the titles and abstracts. The overall agreement was then calculated using Cohen’s kappa statistic . Full teThe Joanna Briggs Institute Data Extraction Form for Economic Evaluations was used to extract data from the included studies . The priData items extracted included descriptive data about the study and analysis including (i) study population/participants, intervention, comparator(s) and outcomes; (ii) study methods including prices and currency used for costing, time period, sensitivity analyses and measures of resource use; (iii) study context ; (iv) analysis methods.Results for the resource use and/or cost and/or cost-effectiveness measures and the author conclusions were also extracted.Critical appraisal of studies was undertaken using the Joanna Briggs Institute Critical Appraisal Checklist for Economic Evaluations , adaptedData extracted from included studies were analysed and synthesized in a narrative summary to address the stated review objectives. No meta-analysis was conducted due to significant heterogeneity of service configurations in the included studies.The study selection process is presented in Fig. Table Ten (53%) of the included studies were conducted in the United States, three in the Netherlands, two in Canada, two in Australia, one in Germany, and one in the United Kingdom. Ten of the studies involved interventions pertaining to processes of care, while nine examined structures of care. Six studies identified examined dementia-specific service configurations.Study designs were varied. The most frequent study design was a cluster-randomised controlled trial (7/19), followed by cross-sectional (3/19), randomised controlled trial (2/19), and quasi-experimental (2/19). Other study designs included controlled before-and-after, non-randomised experimental trial, prospective cohort, retrospective cohort, and a Markov simulation model.The number of participating facilities per study ranged from 1 to 177 . Thirteen of the studies recruited resident participants, with sample sizes ranging from 44 to 6663 , while five studies assessed facility-level data only.Table In a study evaluating a multidisciplinary integrated care model, MacNeil Vroomen and colleagues  also choIn terms of the reporting of resource use and costs there were notable deficiencies in a number of studies. Six out of 19 of the included studies did not disclose the date for their cost data collection , 23–27. Five studies conducted sensitivity analyses , 31–33. Table Four studies evaluated the costs and effects of enhanced staffing levels, including increasing the amount of direct nursing care time for each resident , employiAnother important finding from this review was the assimilation of currently available evidence relating to the costs and effectiveness of staffing levels in specialised models of residential care, including Green House facilities and dementia special care units , 36, 37.Three out of four studies which evaluated staffing levels in specialised models of care  reported that these types of specialised models generally provided more direct care time to residents compared to traditional facilities , 36, 37.One study evaluated the implementation of an evidence based staff education and best practice program targeting ‘vision awareness’ to improve staff knowledge of visual impairments and to reduce the incidence of falls . It was Table Four studies evaluated dementia-specific care interventions compared to usual care. These interventions included person-centred care implemented through staff training , 32, 39 Supporting personhood has been identified as a foundation for quality care for people living with dementia . Person-Two methods of implementing person-centred care were evident from the identified studies. One method, which researchers called ‘person-centred care’ involved off-site staff training followed by a period of on-site supervision and telephone support , 39. TheThe most common primary outcome assessed in this subgroup was agitation using the Cohen Mansfield Agitation Inventory (CMAI) , 31, 39.Other outcomes assessed (and measurement tools used) across this subgroup included emotional responses in care (ERIC), quality of life , care interaction quality , psychiatric symptoms (neuropsychiatric inventory), behavioural symptoms , antipsychotic drug and restraint use, cognition , level of nursing care , and activity participation rates. Some small improvements were found in quality of care interactions, resident care responses, and quality of life measured with the DemQol-proxy .Rovner and colleagues  evaluateTwo studies evaluating integrated care delivery found higher costs in the intervention group compared to usual care , 22. IntPaulus and colleagues  examinedMacNeil Vroomen and colleagues’  integratFour studies conducted facility-level interventions aimed at improving the quality of care , 33, 43.In comparison with the health care sector, where economic evaluations are common practice for pharmaceuticals and medical technologies, this review identified a paucity of economic evidence relating to the structures and processes of care in the residential aged care sector. A total of 19 studies were identified by this review: 12 cost analyses, one cost-minimisation analysis, one cost-utility analysis, two cost-effectiveness analyses, and three cost-benefit analyses.Despite the heterogeneity of interventions and outcome measures, synthesis of study results revealed several common themes. Results from three studies suggest a potential for cost savings to the health care sector by increasing the amount of direct care time provided to each resident , 28, 35.Interestingly, increased levels of direct care time were found in the small, home-like ‘Green House’ model, as well as the dementia special care units. While we would expect to see cross-sectoral benefits  similar to those reported in the enhanced staffing interventions, none of the studies actually measured costs in the health care sector. Three of the four did not report any effectiveness measures , 36, 37,Another aspect of residential care that was shown to create cost savings from a broader health care perspective were quality improvement initiatives, such as activity programs and interventions aimed at reducing health care utilisation and hospitalisations. While quality improvement initiatives tend to come at a cost to the facility in terms of planning and implementation, the flow-on effects of improving care quality is likely to extend to other areas of health services. Many of these initiatives, however, such as the quality improvement projects evaluated by Ouslander and colleagues , and RanThe remaining studies are difficult to generalize, largely due to differing implementation methods. In terms of caring for individuals with dementia, recent research into person-centred care suggests its potential to reduce agitation and aggression , 39, thoThe concept of integrated care is not well-defined, and is therefore difficult to generalize. Two studies identified by this review defined integrated care in terms of integration between staff and residents , and intAnother issue affecting the generalizability of findings is the geographic concentration of research in the United States. Research conducted outside of the United States is sparse. More than half of the included studies were conducted in the United States, while the remaining third were split between the Netherlands, Germany, Canada, Australia, and the United Kingdom. While these findings are consistent with a recent systematic review of randomised controlled trials in care homes, which reported that 50% of the randomised controlled trials undertaken in care homes were from the United States , they doAnother important factor to facilitate transferability of findings in residential aged care, and particularly dementia-specific models of care, is the question of the most appropriate primary outcome measure to use in economic evaluation. All of the dementia-specific studies into person-centred interventions used agitation as the primary outcome, and some small but significant decreases were detected for person-centred care and person-centred environments , 39. AgiAcknowledging that the economic evidence of program features which directly relate to how care is provided in terms of the workforce and its operations (structures of care) and the services provided (processes of care) is limited, we have selected a number of recommendations for change based on the best evidence available. Firstly, increasing the amount of direct care time provided to each resident appears to have wide-ranging benefits at both an institutional and health care level. While further research is needed, additional direct care time provided by nurses, allied health professionals, and other aged care workers all appear to provide benefit. Secondly, benefits arising from initiatives such as increased direct care time or quality improvement initiatives are likely to occur in the health care sector rather than the aged care sector. Future research and policy decisions surrounding residential care initiatives should strive to include health care costs and benefits when considering resource allocation decisions.In terms of methodological recommendations, our primary suggestion is improved transparency in reporting study methods and results. Future economic evaluations in this area should strive to meet the quality standard for reporting economic evaluation as specified in the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) statement  includinThis systematic review has several limitations. Firstly, the search strategy was restricted to English-language publications, which may have resulted in some relevant international research being excluded. Secondly, due to the large number of results retrieved when searching the multidisciplinary database ProQuest, limits to source type were applied that were not part of the original search strategy. The ProQuest search was limited to scholarly journals, reports, dissertations and theses, conference papers and proceedings, and working papers. Newspapers, trade journals, wire feeds, magazines, other sources, books, and encyclopedias and reference works were excluded. While this may have resulted in some relevant research being missed, this limitation was justified to maintain the feasibility of abstract screening within the given time constraints. Finally, due to the broad scope of this review, the synthesis and analysis of results was limited by the heterogeneity of included studies.This review provides the first comprehensive summary of the existing economic evidence pertaining to workforce structures and care processes in residential care, and highlights an urgent need for robust economic evaluations to inform future service development in this area. In order to fully capture the impact of an intervention or model of care in a residential aged care setting, it is important to take a societal perspective when conducting economic evaluations. The inclusion of broader health care costs in economic evaluations of interventions in residential care, in particular the use of hospitals, is critical for ensuring the value of the intervention is not underestimated. Furthermore, the practical application and transferability of findings would benefit from identifying appropriate and meaningful outcome measures that can be used at a service planning level.This review also brings to light the potential value of direct care time for residents in care homes. Future research should explore cost-effective methods for increasing the amount of direct care time to residents, and identification of the most appropriate skill mix  for the provision of care according to the care needs of the individual.Economic evidence is essential to the promotion of efficiency, facilitating future policy directions within the aged care sector and will assist in identifying and quantifying the cross-sectoral impacts of new innovations in the structures and processes of care in terms of both the costs and benefits provided.


Acanthamoeba polyphaga mimivirus (APMV) was discovered in 2003. APMV belongs to the nucleocytoplasmatic large DNA virus (NCLDV) family and infects not only A. polyphaga, but also other professional phagocytes. Here, we review the Megavirales to give an overview of the current members of the Mimi- and Marseilleviridae families and their structural features during amoebal infection. We summarize the different steps of their infection cycle in A. polyphaga and Acanthamoeba castellani. Furthermore, we dive into the emerging field of virophages, which parasitize upon viral factories of the Megavirales family. The discovery of virophages in 2008 and research in recent years revealed an increasingly complex network of interactions between cell, giant virus, and virophage. Virophages seem to be highly abundant in the environment and occupy the same niches as the Mimiviridae and their hosts. Establishment of metagenomic and co-culture approaches rapidly increased the number of detected virophages over the recent years. Genetic interaction of cell and virophage might constitute a potent defense machinery against giant viruses and seems to be important for survival of the infected cell during mimivirus infections. Nonetheless, the molecular events during co-infection and the interactions of cell, giant virus, and virophage have not been elucidated, yet. However, the genetic interactions of these three, suggest an intricate, multilayered network during amoebal (co-)infections. Understanding these interactions could elucidate molecular events essential for proper viral factory activity and could implicate new ways of treating viruses that form viral factories.Viral infection had not been observed for amoebae, until the  Acanthamoeba polyphaga population were at first erroneously classified as bacteria  in 2003  as founding member, which was discovered in 2009  was discovered by incubating water from the Seine river in France with A. castellani, a close relative of A. polyphaga  have been functionally associated with either fibril biosynthesis or as components of the fibrils viridae family that is divided into the Sputnikvirus and Mavirus genera -dependent mavirus  was discovered together with the Mont1 mimivirus in soil samples from Tunisia , lonely member of the Other virophages have been discovered by metagenomic analysis of water samples [e.g., the Organic Lake virophage (Yau et al., Megavirales family. The addition of virophages as parasites of giant viruses, their high abundance in the environment, and the genetic interactions between cell, giant virus, and virophage, suggest an intricate, multilayered network during amoebal co- and super-infections. Future studies of these dynamic interactions could elucidate the inner mechanics of viral factories.Since the discovery of its first member APMV in 2003, new giant viruses are discovered continuously in samples from all over the world and added to the All authors designed the mini review. JD: generated the figures and drafted the text; MH and CH: wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In this study, current geographic information system (GIS)-based methods and their application for the modeling and assessment of mining-induced hazards were reviewed. Various types of mining-induced hazard, including soil contamination, soil erosion, water pollution, and deforestation were considered in the discussion of the strength and role of GIS as a viable problem-solving tool in relation to mining-induced hazards. The various types of mining-induced hazard were classified into two or three subtopics according to the steps involved in the reclamation procedure, or elements of the hazard of interest. Because GIS is appropriated for the handling of geospatial data in relation to mining-induced hazards, the application and feasibility of exploiting GIS-based modeling and assessment of mining-induced hazards within the mining industry could be expanded further. Mining within a sustainable framework is important for the welfare of human beings. However, the mining industry has often been perceived negatively because it can be hazardous to both public health and safety, and cause damage to the surrounding environment, including the land, soil, water, and forests at local, regional, and global levels . Mining-To mitigate or eliminate the potential negative environmental impacts and risks associated with mining activities, an examination of both the extent and the level (degree) of hazards based on continuous assessment within a spatial context is necessary . For thiGIS is a computer-based technology and methodology for collecting, managing, analyzing, modeling, and presenting geospatial data for a wide range of applications . RecentlThe scope of this review paper was confined to published literature concerning GIS-based modeling and assessment of mining-induced soil, water, and forest related hazards that included all the following keywords (or concepts): GIS, mine, type of hazard. For soil-related problems within mining areas, the keywords “soil contamination” or “soil erosion” were used to search and review the literature. Articles concerning soil in urban areas and commercial districts, and soil problems caused by shooting ranges or other activities, were not taken into account. Many researchers have collected samples from soil, stream sediment, and/or water and they have presented elemental concentration data for specific points on a map. However, such studies lack a GIS analytical element and thus, they were excluded from this study. For problems concerning water in mining areas, including tailing dam sites, the terms “water pollution” or “drainage” were used as keywords. The terms “deforestation” or “reforestation” were used in relation to hazards associated with forests in mining areas. Numerous studies have reported on the detection and monitoring of acid mine drainage (AMD), water pollution, and changes in forest area in mining areas using remote sensing observational technologies  or unmanned aerial vehicle (UAV) photogrammetry. However, these phenomena were considered beyond the scope of GIS technology and they were excluded from this study.The lifecycle of a mine typically includes four phases: exploration and feasibility, planning and construction, operation, and closure . In the Potential toxic elements (PTEs) in the soil at mine sites pose a risk to human health because of their potential to enter the food chain via direct ingestion of dust or the ingestion of plants . ConsequWater management and drainage control at a mine site represent important issues regarding control operational processes, hazard prevention, and secondary hazard prevention. Open-pit mines are affected directly by rainfall and, in particular, surface water control is important for such mines in humid regions . MoreoveChanges in forest cover have attracted worldwide attention because of their potential effects on soil erosion, runoff, and atmospheric carbon dioxide levels . Mining As listed in The subject of soil problems in mining areas is generally classified into pollutant transport based on hydrological analysis, geostatistical spatial interpolation for soil contamination mapping, and sediment yield from mine tailing dumps.Pollutant transport in soil is strongly associated with local hydrological characteristics. Thus, the spatial variations of heavy metal enrichments in soils might be related to natural dispersion processes such as leaching by percolating rainwater or mechanical transport in runoff.Hwang and Kim  examinedYenilmez et al.  determinSuh et al.  performeGeostatistical spatial interpolation and simulation methods can be utilized to generate raster grid-cell-based soil contamination maps and to explore the spatial variations of heavy metal pollution. Soil contamination maps enable the natural background levels for an area to be distinguished from the anomalous anthropogenically enriched levels, and to identify areas of contaminated topsoil that require remedial action.Nakayama et al.  quantifiDong et al.  modeled Khalil et al.  assessedReis et al.  combinedAcosta et al.  evaluateYan et al.  estimateLee et al.  compared2 = 0.9997). In addition, this study delineated the extent and calculated the areas with levels exceeding the warning standards and countermeasure standards of Cu from the raster grid-cell-based soil contamination map , soil erodibility factor , slope length and steepness factor (LS), cropping management factor (C), and supporting conservation practices factor (P). Subsequently, by multiplying the maps of these major factors using a map algebra technique, the mean annual rate of soil erosion  was calculated as follows:GIS can be combined with the Universal Soil Loss Equation (USLE) model  to estim−1 and 40.40–187.64 tons year−1, respectively  and attribute data (property-related characteristics of objects). GIS data can be represented in separate layers.Variety of geospatial data: Mining-induced hazard assessment often requires various forms and types of data . Mostly, the sources of these data could be aerial photographs or satellite images, field surveys, and laboratory analyses, and they are usually in the forms of digital maps, tables, and figures.Flexibility of operations and concurrent display: GIS packages or software offer numerous data processing functions, data management tools, and data analysis capabilities in a highly flexible manner with a concurrent interactive display facility.Fast and inexpensive processing: GIS can efficiently store, process, analyze, and visualize large volumes of spatial data, which otherwise would be too expensive, tedious, and time-consuming to conduct by other methods.High accuracy and repeatability of results: The technique is based on digital mapping and it yields higher accuracy compared with manual cartographic products. In addition, the records and results are suitable for rechecking and confirmation.To handle mining-induced geohazard related data, GIS is appropriate for the following reasons:Because GIS can also be considered computer-coded mapology, it can easily represent and combine factor maps and therefore, effectively derive susceptibility, hazard, and risk indices as well as perform modeling efficiently. Therefore, GIS is considered very important in the entire process of mining hazard modeling and mapping production. In addition, GIS-based geohazard mapping based on other concerned geospatial data sets can provide basic data both for engineers and for planners to assist them in making decisions.Although GIS is a useful tool, GIS-based approaches remain challenging to model and assess the mining-induced hazards. The inaccuracies and imprecisions of input data can affect the quality of the resultant map. Most of previous GIS-based studies employed 10–30 m spatial resolution to analyze and mapping mining-induced hazards because it is appropriated for the local or regional scale analysis and mapping (not for site specific). However, aforementioned spatial resolution of the data may have inaccuracies  and is too coarse to precisely model and map the flow, spread, estimation of the actual mining hazards. Therefore, it is necessary to use more high-resolution input data to obtain better results.In this paper, various types of mining-induced hazard and the strength and role of GIS as a geospatial problem-solving tool were introduced. Furthermore, numerous examples of published literature on GIS-based modeling and assessment of several representative mining-induced hazards were discussed. To focus on papers dealing with GIS analysis techniques for the modeling and assessment of mining hazards, remote sensing technology based research was excluded in this study. For the detailed review of the numerous papers, each area of mining hazard research  was classified into two or three subtopics according to the steps of reclamation procedure, or element of hazard of interest .(1) Soil contamination maps provide information regarding hazardous surface soils by representing the distribution of elevated levels of contaminants within the soils. This can assist both in prioritizing the abandoned mines that pose the greatest risk, and in deciding on appropriate remediation measures to prevent the spread of soil contaminants . In this sense, these types of studies are helpful both to engineers and to planners involved in the design and implementation of efficient soil management strategies in abandoned mining areas.To obtain inexpensive and accurate soil contamination maps, efforts to increase the number of sampling sites and to improve the prediction accuracy of interpolation techniques are needed. The ICP-AES method is accurate but expensive, whereas the PXRF method is rapid (low cost) but relatively inaccurate in the measurement of PTEs in soil. Numerous studies have shown high correlation between ICP-AES and PXRF analysis data for most elements present in soil ,20. ThusIt is evident that the prediction accuracy of a soil contamination map could be improved by increasing the number of sampling sites; however, attempts to increase prediction accuracy by improved geospatial interpolation techniques using confined samples should also be considered. The spatial variability of PTEs could be changed easily because they tend to be dispersed or leached by rainfall or by the rise of groundwater level. In particular, several GIS-based pollutant transporting modeling studies have revealed that the dispersion pattern of contaminants is highly affected by the single-flow direction of surface rainwater. Nonetheless, many previous soil contamination studies have not considered the effect of the single-flow direction of surface rainwater in the interpolation process of generating a soil contamination map. This is because of the inhomogeneity of the hydrological tendency and the difficulty of synthetic interpretation. In future work, it will be necessary to consider the hydrological characteristics as part of the geostatistical interpolation process to analyze the spatial pattern of contaminants for better prediction of the distributions of PTEs in the soil at mine sites.(2) GIS-based studies of water pollution in mining areas have progressed gradually. These studies have increasingly taken into account a greater number of variables and they have derived time-series analysis results. In addition, improved simulation accuracies of the fluxes by rainfall or streams have been developed, and the simulations of the resulting phenomena have become more precise. The types of algorithm applied have also diversified and the hazards have been simulated more realistically, providing improved results for drainage design. Nevertheless, further studies are needed. A method for quantitative flood assessment is required for underground mining sites in flooding modeling. There is also an area for improvement in the modeling of the behavior of ground drainage sewers in surface areas. This could be achieved by algorithm improvement and by proposing techniques for using ultrahigh-resolution DEMs derived from UAVs. Moreover, if collaboration with the field of groundwater modeling were achieved, it would improve the temporal simulation of runoff by linking surface and underground water movements.(3) GIS technology can be widely applied to all of the forest reclamation procedures in mining areas such as deforestation assessment, decision support systems for reforestation, and reforestation assessment and design. Although GIS-based decision support systems for forest restoration in mining areas have been proposed, the economic and environmental improvements realized by applying these methods to actual mining sites are insufficient. Changes in the land cover of forest areas in mining sites are associated with other hazards such as landslides, and soil erosion. Therefore, it is necessary to evaluate the use and effectiveness of GIS technology by analyzing the overall process including the evaluation of the degree of forest degradation, application of forest recovery methods, and assessment of the outcome of forest restoration.The widespread use of GIS has significantly improved the modeling and assessment capabilities of mining-induced hazards associated with soil, water, and forest. Furthermore, related research continues to improve and develop via the use of ultrahigh-resolution geospatial data, advancement of spatial data analysis techniques, and coupling of GIS technology with various  models and methods. However, a gap is present between the derivation of maps (by researchers) and their practical use in mine reclamation and management works . Consequently, greater efforts are required to minimize this gap by maximizing the applicability and practicality of future modeling and assessment results.


In the developing vertebrate embryo, segmentation initiates through the formation of repeated segments, or somites, on either side of the posterior neural tube along the anterior to posterior axis. The periodicity of somitogenesis is regulated by a molecular oscillator, the segmentation clock, driving cyclic gene expression in the unsegmented paraxial mesoderm, from which somites derive. Three signaling pathways underlie the molecular mechanism of the oscillator: Wnt, FGF, and Notch. In particular, Notch has been demonstrated to be an essential piece in the intricate somitogenesis regulation puzzle. Notch is required to synchronize oscillations between neighboring cells, and is moreover necessary for somite formation and clock gene oscillations. Following ligand activation, the Notch receptor is cleaved to liberate the active intracellular domain (NICD) and during somitogenesis NICD itself is produced and degraded in a cyclical manner, requiring tightly regulated, and coordinated turnover. It was recently shown that the pace of the segmentation clock is exquisitely sensitive to levels/stability of NICD. In this review, we focus on what is known about the mechanisms regulating NICD turnover, crucial to the activity of the pathway in all developmental contexts. To date, the regulation of NICD stability has been attributed to phosphorylation of the PEST domain which serves to recruit the SCF/Sel10/FBXW7 E3 ubiquitin ligase complex involved in NICD turnover. We will describe the pathophysiological relevance of NICD-FBXW7 interaction, whose defects have been linked to leukemia and a variety of solid cancers. The formation of a segmented body plan is a conserved feature of embryogenesis for all vertebrate species. This process leads to the formation of transient embryonic segments, called somites. Somites are precursors of vertebrae and ribs, associated skeletal muscles, and some dermis . The determination front marks the point of intersection of these gradients, where the next prospective somite boundary will form  , glutamic acid (E), serine (S) and threonine (T) residues] and a transactivation (TAD) domain  29–36 EFG-like repeats in its extracellular domain, involved in ligand interaction; (ii) three juxtamembrane repeats , required for extra-intracellular domain interaction (located within the Negative Regulatory Region (NRR); (iii) the intracellular region, including seven ankyrin (ANK) repeats flanked by a PEST  protein complex  cases , splenic marginal zone lymphoma (SMZL), Hadju-Cheney syndrome (Isidor et al., Given the importance of Notch signaling in cell fate determination and cell cycle progression, it is not surprising that aberrations in the pathway lead to cancers and other diseases (Roy et al., in vitro. It remains to be shown if this interaction occurs in vivo, if NICD interacts with any other E3 ligases, how this interaction is regulated and whether it is context-dependent. The FBXW7 null mutant mice exhibit elevated levels of Notch4 intracellular domain and/or Notch1 intracellular domain alongside defects that are in alignment with a variety of roles identified for Notch in different developmental process such as cardiogenesis and vascular development. However, intriguingly, with respect to the segmentation clock, the absence of Fbxw7 seems to play a less major role in this process, at least according to the mutant phenotypes—although a detailed analysis has yet to be conducted (Tetzlaff et al., Considering the variety of pathological conditions associated with alterations of NICD and FBXW7, there is a limited understanding of the regulation of this interaction. Our current understanding stems from a study on Sel-10, the nematode homolog of Fbxw7, showing the two proteins bind directly to each other and FBXW7 negatively regulates Notch signaling (Hubbard et al., In this review we provided a general overview of the critical role of Notch signaling in regulating the segmentation clock involved in somitogenesis. Notch activity is based on stability and turnover of its intracellular domain, NICD. This stability is regulated by phosphorylation of the PEST domain, targeting NICD to proteasome degradation upon recognition by the E3 ligase FBXW7. Mutations in the PEST domain, leading to aberrations in NICD stability, are the underlying cause of a number of solid and non-solid cancers and different genetic disorders. Therefore, uncovering the finer details of Notch pathway regulation merits attention, particularly because a wider comprehension of this process would provide further insights into the mechanisms involved in the onset of Notch-related diseases.FC and JD conceived the structure and content. FC wrote the initial draft document. FC designed and produced the figures. JD corrected and edited the document.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In the alarming context of risk of Zika virus (ZIKV) transmission in the Euro-Mediterranean area, there is a need to examine whether capacities to detect, diagnose and notify ZIKV infections in the region are in place and whether ongoing capacity-building initiatives are filling existing gaps.The MediLabSecure network, created in 2014, comprises 55 laboratories of virology and medical entomology and 19 public health institutions in 19 countries in the Balkans, North-Africa, the Middle-East and the Black Sea regions. It aims to set up awareness, risk assessment, monitoring and control of emerging and re-emerging vector-borne viruses. We here examine the actions and strategies that MediLabSecure has been implementing and how they will contribute to the prevention and control of the ZIKV threat in the Euro-Mediterranean area.Capacity-building for arbovirus diagnostics is a major objective of the project and follows a methodological rather than disease-driven approach. This enables the implementation of laboratory trainings on techniques that are common to several arboviruses, including ZIKV, and putting into action appropriate diagnostic tools in the target region.Moreover, by its One Health approach and the interaction of its four sub-networks in human virology, animal virology, medical entomology and public health, MediLabSecure is fostering intersectoral collaboration, expertise and sharing of information. The resulting exchanges  across disciplines and across countries, dedicated research on intersectoral collaboration and increasing diagnostic capacities are providing new paths and tools to public health professionals to face emerging viral threats such as a ZIKV epidemic in the Euro-Mediterranean region. Due to the link between Zika virus (ZIKV) infection and neurological disorders/neonatal malformations, the WHO acknowledged the ongoing ZIKV outbreak as a public health emergency of international concern (PHEIC) –4. At leStegomyia, Diceromyia and Aedimorphus of the genus Aedes) [Stegomyia mosquitoes, including the invasive tiger mosquito Aedes albopictus that is spreading in the Euro-Mediterranean and Black Sea area [Indeed, ZIKV has been isolated from a variety of mosquito species, mainly of the Aedini tribe (among which the subgenera s Aedes) , 10, indSea area , 12, comAedes albopictus mosquito species [Considering this alarming context, the ECDC recently issued a ZIKV risk assessment for continental Europe, estimating an increasing number of travel-related reported ZIKV infections, and indicating a risk of transmission from imported cases by the  species , now est species . This fiIn order to enhance preparedness to common health threats and biosafety risks, the EU supported the creation of the MediLabSecure network of human, veterinary and medical entomology laboratories and public health institutions which aims to support target countries to set up awareness, risk assessment, monitoring and control of emerging and vector-borne viruses of concern in the region. This network was created in 2014 as a continuation of a prior network developed by the EpiSouth and EpiSouth Plus Projects , 16 on tThis One Health project rotates around the interaction of four sub-networks: one for human health, one for animal health, one for medical entomology and one for public health. This approach targets viral threats that can have a great impact on human and/or animal health.The purpose of this correspondence is to present the actions and strategies that MediLabSecure is implementing and how they can now contribute to the prevention and control of the ZIKV threat. We aim to highlight how, although established prior to the current crisis, MediLabSecure has been implementing a number of WHO recommendations included in the ZIKV PHEIC Declaration  under a The overall MediLabSecure objectives are: (i) identify the needs of the different target laboratories by assessment tools, (ii) design and implement tailored training sessions accordingly, (iii) complement with expert visits and quality assessments and (iv) strengthen intersectoral collaborations between the human, animal and entomological sectors involved in vector-borne diseases surveillance. This objective is addressed by holding multidisciplinary “One Health” activities such as joint meetings, technical transversal workshops, expert visits where representatives of the four networks interact at national and international levels to foster mutual cooperation and intersectoral risk assessment exercises and tailored research to explore how this intersectoral collaboration is implemented within partner countries.human virology sub-network, under the leadership of the Institut Pasteur in Paris, conducted three workshops on scientific knowledge and technical skills  for the detection of arbovirus infections. While specific diagnostic capacity for ZIKV in terms of reagents and reference protocols was not initially an aim of this project, the MediLabSecure human virology sub-network reacted to this new public health context and, by January 2016, ZIKV real-time RT-PCR protocols were shared and required reagents including positive controls were distributed within the network. Information regarding the availability and performance of diagnostic tools and reference protocols as well as accurate testing algorithms are shared and regularly updated.To date, the MediLabSecure animal virology sub-network, under the leadership of the Animal Health Research Centre , is working to enhance preparedness of veterinary laboratories to face health threats caused by emerging viral pathogens, most of them zoonotic in nature. Technical workshops, experts’ exchanges, scientific visits and external quality assessment exercises are performed to improve the capacities of the laboratories to diagnose zoonotic viral infections in animals, with special focus on arboviruses. Finally, the overall objectives are completed by promoting continuous contact and exchange of information not only between veterinary laboratories from the different participating countries, but also within the different sub-networks, in-line with the adopted “One Health” perspective.The MediLabSecure Although ZIKV is not regarded as a zoonotic agent, there are still a number of uncertainties around its host range and life cycle. For instance, there is evidence that bats could play a role in ZIKV infectious cycle . More stmedical entomology sub-network, under the leadership of the Research and Development Institute , focuses mainly on mosquitoes identified as present threats for arbovirus transmission but also with potential risk of emergence in the Mediterranean and Black Sea regions. Expert visits, and training modules have been implemented and mosquito identification tools have been devised. Three one-week training sessions have enabled at least two entomologists from each of the 19 laboratory members to increase practical competencies concerning mosquito vectors of arboviruses  and to enhance regional cooperation.The MediLabSecure public health sub-network designed a framework to define, study and promote intersectoral integration of surveillance from a One Health perspective through a systematic literature review, a survey involving all the sub-networks in all 19 countries [In view of strengthening integration across disciplines for surveillance of vector-borne diseases, the MediLabSecure ountries  and the The current ZIKV emergence has highlighted the need to conduct comprehensive risk assessments, establish effective monitoring programs, develop preparedness activities and implement adequate control measures with regard to arboviral threats in the Euro-Mediterranean area. Thanks to its One Health framework integrating four sub-networks involved in the monitoring, prevention and control of arboviral diseases from different perspectives, MediLabSecure is fostering collaboration, networking and timely sharing of information, across disciplines and across countries. In fact, as also discussed by other authors , the desThe resulting exchanges and capacity-building are now facilitating the step-up of intersectoral regional capacities in the face of emerging viral threats in a methodological rather than disease driven approach. Therefore, after having developed the basic methodological skills across the region, the network is now in the position to fill gaps in ZIKV detection capacity, by putting into action harmonized diagnostic tools and algorithms, vector sampling and identification methods and control measures, within a conducive intersectoral framework which constitutes an asset towards preparedness to any present or emerging threats to human and/or animal health.


Never has this expression been truer than in the clinical and pharmaceutical arenas. Enormous strides have been made by the scientific community in the evolving field of biomedical imaging with the aim of representing and/or quantifying aspects of disease and drug action by using tools such as radiography , MRI 2,,2, 3 PETImage analysis has been a focus of mass spectrometry for more than 40 years since early studies using secondary ion mass spectrometry (SIMS) . Among tThe pharmaceutical industry has taken advantage of the development of IMS to enable an array of high-throughput screening modalities for pharmaceutical assessments . IMS canAt this 1-day symposium, the Department of Environmental Health Sciences (DEHS) at the Yale School of Public Health brought together leaders in IMS to discuss recent developments, limitations, and future needs, and to increase awareness of this growing and important field. During his opening remarks, Dr. Vasiliou highlighted the potential of IMS to define the molecular basis of diseases, to provide insights into mechanisms, and to integrate tissue morphology at the molecular level. Thereafter, talks by an impressive assembly of thought-leaders in IMS illustrated the potential of unbiased tissue imaging to deliver a new level of understanding of pathophysiological processes at the molecular level. The symposium concluded with a round-table discussion, chaired by Dr. Mark Duncan, on some of the more practical issues in IMS such as current bottlenecks and future opportunities. The presentations and discussions at the symposium underscored the great potential of IMS. With the intention of bringing IMS to the larger scientific community, the DEHS has committed significant resources to the acquisition of equipment and expertise that will allow the further development and application of IMS techniques.The first talk of the symposium was given by a pioneer of the field, Dr. Richard Caprioli, Professor of Biochemistry and Director of the Mass Spectrometry Research Center at Vanderbilt University. His opening remarks gave a historical overview of MALDI IMS technology and emphasized its advantages. Dr. Caprioli explained how MALDI IMS employs desorption of molecules by direct laser irradiation to map the location of specific molecules from fresh frozen or formalin-fixed tissue sections without the need to target specific reagents, such as antibodies . Dr. CapAldh3a1-null mice presented the first genetic animal model of cellular-induced corneal haze due to the loss of a corneal crystallin [Dr. Kevin Schey, Professor of Biochemistry, Ophthalmology and Visual Sciences, at Vanderbilt University, discussed the application of IMS to study a range of molecular classes, such as proteins, lipids, and metabolites in ocular tissues. Ocular tissues provide an ideal medium to demonstrate the utility of the technique where morphological features are on the scale of single cells. For example, molecular profiles can be produced in retina pigment epithelium. Moreover, a range of diseases affect the various ocular tissues, including glaucoma, age-related macular degeneration, cataract, and corneal cataract. Dr. Schey illustrated how IMS is being actively applied to derive mechanistic information that enhances understanding about the molecular underpinnings of disease in these tissues as well as aging mechanisms. IMS data from optic nerve, retina, lens, and cornea were presented with special attention to diseases affecting these tissues . Data frystallin . This woén, Professor at Uppsala University, who showed novel ways to interrogate the actions of neurotransmitters, their precursors and metabolites, in the brain chemical network and neuronal signal transmission [én uses knowledge about the relative abundance and spatial distribution of neurotransmitters in the brain to provide insights into these complex neurological processes and disorders. At present, researchers rely on indirect histochemical, immunohistochemical, and ligand-based assays to detect small-molecule transmitter substances or on tissue homogenates analyzed by high-performance liquid chromatography analysis. Current neuroimaging techniques have very limited capacities to directly identify and quantify neurotransmitters from brain sections. MALDI IMS can perform analyses directly on the surface of a tissue section, establishing itself as a powerful in situ visualization tool for measuring abundance and spatial distribution of endogenous and pharmaceutical compounds, lipids, peptides, and small proteins. A novel reactive MALDI matrix, recently developed by Dr. Andrén’s group, selectively targets the primary amine group on neurotransmitters, metabolites, and neuroactive substances while also functioning as a matrix to enable ionization [én’s laboratory has therefore developed a new reactive matrix that can selectively target and charge-tag both phenolic and primary amine groups, thus enabling MALDI IMS of both MAO and COMT downstream metabolites, focusing on a nucleophilic aromatic substitution reaction with such functional groups. Using this new reactive matrix, they were able to detect and map the localization of most of the neurotransmitters and metabolites involved in the dopaminergic and serotonergic network in a single brain tissue section. This work showed a novel methodology that assists with metabolite identification through the selectivity of the reaction. The sensitivity and specificity of this imaging approach to neurochemicals has great potential for many diverse applications in neuroscience, pharmacology, drug discovery, neurochemistry, and medicine.The broad range of IMS applications was reinforced even more by Dr. Andrsmission . Changesnization . HoweverA major focus of the symposium was the application of MALDI IMS to map the distribution of a variety of therapeutic molecules across a tissue section of interest and to assess their biological impact.Current president of the Imaging Mass Spectrometry Society and director of US Imaging MS, at GlaxoSmithKline (GSK), Dr. Castellino, discussed how MALDI IMS technology has taken their research beyond “plasma-centric” studies and allowed for direct mapping of molecular changes in tissue associated with drug pharmacology, disposition, and disease pathogenesis. Delivering safe and efficacious drugs is tied to the ability to understand complex mechanistic relationships between molecular initiation events of pharmacologically active compounds and the cascade of subsequent biological consequences. Because the delivery of drugs to their intended target, and avoidance of unintended targets, is a critical first step, IMS can directly guide improvements and innovation in delivery strategies by mapping the target tissue selectivity . FurtherDr. Richard Goodwin, a principal scientist for Drug Safety and Metabolism at AstraZeneca (AZ) and head of mass spectrometry imaging, presented the challenges faced for drug discovery and development; it is a lengthy, high risk, and competitive business that can take a decade to progress; moreover, billions of dollars are required to move a new medicine to market. Dr. Goodwin discussed how IMS could help mitigate some of the primary reasons for drug attrition, specifically around lack of efficacy and toxicological or clinical safety risk. IMS is now demonstrating impact on drug discovery programs and helping reduce later stage compound attrition. It provides insights into the biodistribution of compounds, while simultaneously generating data on pharmacodynamic biomarkers. Dr. Goodwin presented data from AZ that showed how the use of a range of multimodal imaging techniques improves understanding about compound efficacy, safety, and targeted drug delivery , 33. InvDr. Sheerin Shahidi-Latham, Head of Metabolomics and Imaging MS, Department of Drug Metabolism and Pharmacokinetics at Genentech Inc., also discussed the advantages of the applied use of MALDI IMS. In the pharmaceutical industry, obtaining information about the absorption, distribution, metabolism, and elimination (ADME) of a new chemical entity via a PK study in a preclinical animal model is often the first step towards understanding the in vivo properties of a drug-like molecule in humans. Traditionally, much of this ADME work has been supported by liquid-chromatography coupled to mass spectrometry. In the case of tissue distribution, the organs are excised and homogenized in order to accommodate this analytical workflow, thereby effectively eliminating any spatial information. Dr. Shahidi-Latham emphasized that MALDI IMS has gained prominence since it provides a robust, label-free detection of drug and metabolites while preserving spatial localization within tissue sections of interest. Additionally, the use of high-resolution mass spectrometers has provided the opportunity for simultaneous detection of subsequent pharmacodynamic (PD) responses within a single image acquisition. Dr. Shahidi-Latham discussed how the ability to assess PK/PD relationships in a label-free, in situ context has proven invaluable to the early lead optimization efforts that take place in the drug discovery setting. Similarly, uncovering the perpetrator of adverse effects often associated with histopathological assessments in the preclinical development phase has also improved their understanding about the mechanisms of toxicity and can provide useful information for the redesign of a back-up molecule. The presentation provided a synopsis of the advantages of IMS, as well as the technical challenges and opportunities in the context of the pharmaceutical industry. Dr. Shahidi-Latham presented data from her work at Genentech, which included MALDI IMS of dosed tissues in support of drug efficacy, PK/PD, and effective delivery evaluations, and highlighted the simultaneous detection of drug, metabolites, and endogenous components attainable from a single imaging run –39. MoreThe constant drive towards integrating complex and large-scale datasets has generated the need to develop new tools to process this information . Dr. KirThe symposium concluded with a round-table discussion, chaired by Dr. Mark Duncan, where the attendees discussed the practical challenges and future directions of IMS. The attendees not only agreed on the potential of IMS to provide previously inaccessible insights into molecular events at the tissue level, but also highlighted the cost and complexity of both the science and the technology that underpins these studies. Rather than a routine core service, it was agreed that designing meaningful studies, performing exacting sample handling, generating and interpreting complex data, and maintaining high-end instrumentation requires a substantial, highly collaborative interaction between all stakeholders.


Howeverem (SNS) , 12. ThiThe concept of leptin resistance is used to define states where hyperleptinemia is combined to lack of response to the hormone, with consequent maintenance of body weight excess and increased food consumption . Neverthvia activation of beta-adrenergic receptors post-synapticaly in target organs  and elevation of risk for developing cardiovascular diseases . MoreoveMost Obesities kNown Are Low In Sympathetic Activity). This hypothesis was built up from studies of SNS activity and norepinephrine decay in obese patients and rats (tabula rasa models precede the era of molecular genetics, which paved the way for molecular and functional diversity of seemingly alike cells. Indeed, the era of molecular genetics has enabled the mechanistic dissection of brain circuits, as well as, the immune system in spectacular ways. However, the molecular and circuit organization of the SNS, which innervates all known organs, is essentially unexplored. A molecular neuroanatomical map of the SNS may in turn revive the MONA LISA hypothesis. Once we understand the diversity of the SNS from a molecular and circuit standpoint, we then may be closer to resolving the mistery of leptin resistance.Taking into account all the aspects referred above, leptin resistance may also involve decreased sympathetic local activity, within the adipose organ. In this regard, and before the leptin era, George Bray and others proposed the “MONA LISA” hypothesis (and rats –46. Howeand rats . The conAll authors listed have made substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Significant progress has been made in the management of renal cell carcinoma (RCC) during the last few decades. In early stage, localized disease, surgical resection remains the modality of choice, with no therapeutic interventions as options for post-operative therapy other than simple observation and clinical surveillance. However, treatment options in the advanced or metastatic setting are increasing at a dizzying pace, initially with cytokine therapy, then with the increased availability of targeted therapy including novel small-molecule inhibitors of receptor tyrosine kinases and monoclonal antibodies targeting novel proteins, establishing them as the current standard of care. Even more recently, immunotherapy has seen tremendous development in the form of immune checkpoint inhibition and vaccines. Overall, these interventions have gradually changed the landscape of cancer management in general, and metastatic renal cell carcinoma (mRCC) in particular. This is exemplified by the recent United States Food and Drug Administration (USFDA) approval of nivolumab for patients with mRCC after failure of TKI therapy. In this review, we present a brief overview of the current management of mRCC, primarily the clear cell subtype (ccRCC), and discuss the major clinical trials and data on the immunotherapy in advanced or mRCC. However, no adjuvant treatment has been proven to be beneficial. A significant proportion of patients eventually develop tumor recurrence, and subsequent therapeutic options in the advanced or metastatic setting are limited. Since the turn of the millennium, novel small-molecule targeted therapy has evolved to become the mainstay of treatment for metastatic renal cell carcinoma (mRCC), though prognosis remains poor  , pidilizGiven the success and promising data involving checkpoint inhibition in metastatic RCC, it is reasonable to ask the question whether immunotherapy plays a role in the non-metastatic adjuvant setting. However, various randomized trials of adjuvant therapy involving tumor cells plus BCG, tumor cell vaccination, IFN-α, high-dose IL-2, or a combination of cytokines have not demonstrated survival benefit when compared with observation . The rolRCC is often considered to be a radio-resistant disease and the role of radiation is generally limited to palliative or local control. Preclinical data from the 1990’s from a RCC mouse model demonstrated the regression of lung metastases with radiation and an even greater degree of decrease when combined with IL-2 . In addiThe role of immunotherapy in the management of RCC has changed dramatically since the turn of the century. Previously, cytokine therapy involving IL-2 and IFN-α was the mainstay of mRCC treatment. However, benefit was only marginal considering the toxicities related to therapy. Over the past decade, significant progress has been made in the development of new immunotherapeutic agents in the form of checkpoint inhibition and vaccines, and these agents are gradually being incorporated in the treatment of mRCC and many other cancers, such as melanoma, non-small cell lung cancer, Hodgkin’s disease, and head and neck cancer. In mRCC, various checkpoint inhibitors are currently being investigated for use as a first-line agent, following recent USFDA approval of nivolumab in the second-line setting. As the use of immunotherapy becomes more widespread in oncology, many questions will certainly need to be addressed. For example, the long-term side effects and resistance mechanisms of these drugs in various tumors are important considerations. Optimizing treatment dose and schedule, investigating biomarkers that may affect disease outcomes, and evaluating the combination of different treatment modalities are also critical in maximizing the potential of immunotherapy . FurtherIt is now quite clear that immunotherapy is here to stay and has begun to impact the clinical outcome of patients with mRCC. Further enhancements to this efficacy paradigm can be achieved with the use of combination therapies, including with other checkpoint inhibitors and with anti-VEGF TKI. As alluded to earlier in this review, there is strong pre-clinical rationale for combining immunotherapy with anti-VEGF inhibitors. The latter, in particular, have been shown to reduce myeloid derived stem cells (MDSC) that are crucial in tumor-induced immune suppression. Furthermore, development and clinical implementation of predictive biomarkers will be crucial to the use of immunotherapy. Preliminary research, especially the expression of PD-L1 by IHC in the tumor cells, has so far not lived up to the promise of a predictive biomarker . One of In this review, we have sought to highlight the entire field of clinical development of immune based therapy for mRCC, and hope that the reader will find this piece of work useful as a summary and as a source of reference. We also recognize that it is a highly dynamic and evolving field, and the pace of development is astounding and encouraging. The use of immune based therapy for this disease, while decades old (in the form of INF and IL-2), offers new promise with the targeting of the checkpoint pathways. The latter field is still in its infancy stage, with tremendous potential for the future. We believe that a major paradigm shift in RCC management will only occur once we can increase the cure rates of surgically managed patients by finding a drug that will improve survival rates in the adjuvant setting. Such trials using TKI and anti PD-1/PD-L1 agents are in progress, and the results are eagerly awaited.


It has been experimentally shown that host-microbial interaction plays a major role in shaping the wellness or disease of the human body. Microorganisms coexisting in human tissues provide a variety of benefits that contribute to proper functional activity in the host through the modulation of fundamental processes such as signal transduction, immunity and metabolism. The unbalance of this microbial profile, or dysbiosis, has been correlated with the genesis and evolution of complex diseases such as cancer. Although this latter disease has been thoroughly studied using different high-throughput (HT) technologies, its heterogeneous nature makes its understanding and proper treatment in patients a remaining challenge in clinical settings. Notably, given the outstanding role of host-microbiome interactions, the ecological interactions with microorganisms have become a new significant aspect in the systems that can contribute to the diagnosis and potential treatment of solid cancers. As a part of expanding precision medicine in the area of cancer research, efforts aimed at effective treatments for various kinds of cancer based on the knowledge of genetics, biology of the disease and host-microbiome interactions might improve the prediction of disease risk and implement potential microbiota-directed therapeutics. In this review, we present the state of the art of sequencing and metabolome technologies, computational methods and schemes in systems biology that have addressed recent breakthroughs of uncovering relationships or associations between microorganisms and cancer. Together, microbiome studies extend the horizon of new personalized treatments against cancer from the perspective of precision medicine through a synergistic strategy integrating clinical knowledge, HT data, bioinformatics, and systems biology. Our body is integrated by a legion of microorganisms that coexist in all our tissues and, notably, with a symbiotic functional purpose. Furthermore, host-microbial interactions are beginning to be recognized for their outstanding influence on well-being or the emergence of diseases such as cancer. The advent of high-throughput (HT) technologies has allowed significant advancements in uncovering these correlations through the diversity and abundance of microorganisms in samples of normal and dysfunctional cohorts of human tissues associated with complex diseases such as obesity, type 2 diabetes, and cancer. For instance, in 2015, Mitra and co-workers reported the characterization of the microbiota at different stages of development of cervical intraepithelial neoplasia, and they observed a strong association between the severity of the disease and the vaginal microbiota diversity  and Metabolomics, and the construction of quantitative schemes capable of influencing clinical decisions in precision medicine.In this review, we present the forefront of HT technologies and conceptual schemes in bioinformatics and systems biology for surveying the host-microbiome association and cancer progression. We expect that our review will be used as a technical and conceptual guide in human microbiome studies, present and discuss the advances in the field, and establish an introspective analysis of the next steps for linking microbiome studies and precision cancer medicine.The advent of HT technologies has positively impacted the elucidation of the metabolic and regulatory mechanisms by which hosts and microbes interact to determine a health or disease state in the host. In particular, NGS and techniques related to metabolome analysis such as mass spectrometry (MS) are valuable technologies for analyzing the microbiota composition and exploring the genetic, functional, and metabolic activity of the microbial community. Moreover, the use of these technologies enables us to explore the implications of the human microbiome to induce functional and dysfunctional states in a variety of human tissues. Here, we present the state of the art of these technologies and discuss some key findings to elucidate the relationship between the human microbiome and cancer.Sanger sequencing, the first-generation of DNA sequencing technology developed by Frederick Sanger based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase, established the methodological principles for DNA sequencing  and the lowest error rates compared with 454 GS Junior or Ion Torrent PGM  marker gene sequencing community identification, which surveys and counts microbes using amplicon sequencing of a single marker gene that is usually the 16S rRNA gene, and taxonomic assignment by bioinformatic methods; and (b) shotgun metagenomic sequencing, which surveys the entirety of all microbial DNA present in a sample using a collection of s Brown, .in vitro. For that reason, their full genetic spectrum was unknown until the advent of HT sequencing technologies, expanding our knowledge of the microbial world. The similarities and distinctions among bacterial species have become complex  regions are amplified using particular sets of primers. The choice of which variable region to use and amplify depends on factors related to the sample and experiment. For instance, evidence suggests that the V1–V3 region is better for taxonomical classification of species; however, some predictive studies show that the V3–V5 region results in a better classification of microbiota from disease vs. healthy specimens , allows for the analysis of all of the RNA of a sample to determine which genes are transcribed and for monitoring gene regulation over time, which is particularly interesting when studying changes in the microbiota in response to perturbations  or the FASTX-Toolkit, which is a collection of command line tools. Parameters for good quality data include a Phred quality score above 28, low percentage of duplicated sequences, no adapter content, and GC count per read close to the theoretical distribution. Another useful tool for quality assessment and processing of HT DNA sequence data is the Bioconductor's package ShortRead  and De Bruijn Graph (BG). OLC methods are based on overlap graphs, and their process has three steps: (1) searching for overlapping reads comparing all-against-all, (2) construction and manipulation of an overlap graph leading to an approximate read layout, and (3) constructing the consensus sequence using multiple sequence alignments  by alignment methods , FASTA  or HMMER (hidden model Markov profiles) when sequence identity is low. These analyses can be performed using locally installed software; alternatively, for users with no bioinformatic training, there are different suites for analysis, such as MG-RAST , two different databases can be used: KEGG  fingerprinting or endo-metabolome, searching for metabolites within the organisms under study; 2) footprinting or exo-metabolome, analyzing metabolites from the environment around the organism under study; and (3) metabolome profiling, where the goal is to screen one or more specific compounds . MS is a highly sensitive method for detection, quantification and structure elucidation of hundreds of metabolites. Given the wide spectrum of molecular weights of metabolites in samples, it is necessary to separate metabolites to improve the sensitivity and accuracy of detection. Thus, MS is often coupled with different separation techniques such as gas chromatography (GC-MS), liquid chromatography (LC-MS) and capillary electrophoresis (CE-MS) .On the other hand, NMR spectroscopy is a technique with high reproducibility and is able to absolutely quantify metabolites using a single reference; because it is a non-destructive technique, the samples can be used for re-analyses using other methods   while others are associated with potentially detrimental effects  . By using genomic approaches, some studies have compared the mucosal surface and the intestinal lumen microbiota between healthy patients and those with CRC  Jobin, . This su) Jobin, .In vitro studies have reported a signaling process between bacterial quorum-sensing peptides (QSPs) and cancer cells. Bacillus derived QSPs are synthesized when there are bacterial stressors and are able to induce tumor cell invasiveness in a process called epithelial-mesenchymal-like (EMT-like) process (involved in CRC metastasis) , Clonorchis sinensis (C. sinensis), Opisthorchis viverrini (O. viverrini), and Schistosoma haemotobium (S. haemotobium). Although HIV does not directly cause cancer, its infection strongly increases the incidence of many different human cancers. Among these cancers, those associated with the herpesviruses KSHV and EBV are the most strongly enhanced by immunosuppression  of the 12.7 million new cases in 2008 attributable to infection. This fraction is substantially higher in less developed regions of the world  than in more developed regions (7.5%). The most important infectious agents are The IARC estimates that one in five cancer cases worldwide are caused by infection, with most being caused by viruses  and hepatitis C virus (HCV) is known to cause hepatocellular carcinoma , and its pathogenicity island (cag PAI), and vacA (vacuolating cytotoxin A) are the major pathogenic factors  responses in patients with distal gastric adenocarcinoma , host-related factors (gene polymorphisms and immune system status) and environmental aspects   that function as detergents that facilitates digestion and absorption of lipids; while also acting as signaling molecules related to metabolic homeostasis  or Lithocholic Acid (LCA) causes extensive DNA damage at physiological concentrations in a dose-dependent manner  in hepatic stellate cells , mainly acetate, propionate, and butyrate , butyrate induces hyperacetylation of core histone proteins (H3 and H4) when compared with other SCFA. Among its effects as an HDAC inhibitor, butyrate can induce Interestingly, those effects depend on cell status, i.e., normal vs. cancer. In the former, butyrate stimulates proliferation (functioning as an energy source); while in cancerous cells, butyrate inhibits proliferation and induce apoptosis  and DNA mismatch repair (MMR) genes (as commonly observed in humans), Belcheva et al. observed an anomalous proliferation of colonic epithelial cells and formation of polyps  leucine, valine, and isoleucine. This generates a diversity of end products including ammonia, SCFA, and branched-chain fatty acids (BCFA) valerate, isobutyrate, and isovalerate. Microbial metabolism of amino acids can also produce biogenic amines by decarboxylation of amino acids  abundance in human colorectal cancer  in tumors with a high +T cells can produce cytokines that promote tumor progression  mice. The enterotoxigenic B. fragilis induces STAT3 signaling characterized by a selective TH17 response for colonic hyperplasia and tumor formation , and this metabolite has proatherogenic properties  the coherent interpretation of heterogeneous HT data; (2) identification of potential biomarkers in cancer; and (3) the optimal design of personalized treatments in clinical trials  is an emergent property that results from host-microbiome interactions. Understanding how this property emerges at a molecular level is valuable to reach one of the aims in precision medicine: the desire for more effective treatments in cancer based on personalized genetic background and lifestyle. In this context, HT technologies and biochemical, physiological and clinical data can be organized and evaluated using a network approach that can be useful for predicting disease expression or response to therapies . Moreover, in melanoma patients who responded to anti-CTLA-4 treatment, the abundance in the stool of Bacteroides thetaiotaomicron and B. fragilis correlated with the response to therapy. This protective effect can be transferred to mice by a fecal microbial transplant (FMT) .Finally, given the social, ethical and legal implications of personalized information, it should be stored in a protected way. To this end, the management system can include protocols for prevention and protection, access control and a plan of action to prevent the loss of information when some event endangers the integrity and security of the data . In addition, ambitious projects in precision medicine need to leverage important resources, such as research cohort biobanks for longitudinal research studies, and an efficient bioinformatics system that aids in the translation from biomedical research to molecular targeting and identification of biomarkers that correlate with the disease state. Intensive investigations are being conducted to illustrate how microbiome profiles, taking into account relationships with the host, could be used as biomarkers to revolutionize prognostication in cancer.Precision medicine based on powerful HT technologies for characterizing patients, such as genomics, proteomics and metabolomics, and computational tools for analyzing large sets of data will integrate the discovery of biomarkers and the electronic medical records to provide evidence for the improvement of clinical practice. The big challenge of data analysis of HT technologies is the development of new computational algorithms to improve the integration of the information from different platforms. In this context, deep learning and machine learning have been proposed as good alternatives to perform these tasks  can provide more insight on the significance of different compositions in different states (Shade and Handelsman, Finally, precision medicine faces many other challenges that will be addressed not only from a scientific point of view but also from a social and ethical point of view, including the proper distribution of the benefits of these technologies across most regions of the world and the development of reliable computational platforms that allow data to be stored confidentially, private and protected. Likewise, coordination with ethics committees and regulation will also be necessary for the use of information without the risk of infringement of the patient's rights and to understand and regulate the legal, social, and economic implications.All authors contributed extensively to the design and discussions of the material presented in this review. All the authors participated to wrote the final version of the paper.The authors thank the financial support from the Research Chair on Systems Biology-FUNTEL and Instituto Nacional de Medicina Genomica, Mexico.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AA and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


A 49-year-old male who attended our hospital due to a history of left hearing loss over a period of 2 years, associated to otalgia, and vertigo to which bleeding in the left ear and facial paralysis had been added in the last 4 months. He reported the simultaneous onset of a left-sided neck swelling with slow growing. After this, he developed loss of sensation of taste on the posterior third of the tongue and swallowing dysfunction, dysphonia, atrophy of the trapezius and sternocleidomastoid muscle and deviation and atrophy of tongue toward the affected side. On physical examination, the mass was found to be painless, soft, pulsating and semifixed. During the otoscopic examination, a pulsatile erythematous lesion of vascular appearance was noted in the external auditory canal. The audiometric test revealed severe left mixed hearing loss. A Doppler ultrasound was performed, which revealed a hypervascularized tumor in the left-sided neck that extended to the skull base. An axial and coronal computerized tomography (CT) scan was taken, revealing a soft tissue lesion extending from the border of II and III neck region to the middle ear and external auditory canal, causing large jugular bulb, hypoglossal canal and temporal bone erosion. This mass avidly enhanced after CT angiography. Following the results of preoperative examinations, diagnosis of simultaneous left glomus vagale, jugulare and tympanicum was confirmed. As the tumor was unresectable, the patient received fractionated radiotherapy associated with iterative stereotactic radio surgery. The patient has been stable over the last two years.


Long-term potentiation and depression (LTP and LTD) represent powerful means of altering synaptic communication. However, they have an inherent potential to excessively modulate neuronal excitability, possibly resulting in unfavorable destabilization of neural function. It has been suggested that homeostatic regulation  of neuronal networks avoids such destabilization. This construct proposes that the threshold for modification of neuronal excitability is based on previous post-synaptic activity, with an elevated threshold following high neuronal activity, but a reduced threshold following low neuronal activity  to investigate age-related changes in the mechanisms of metaplasticity  (Ziemann and Siebner, GO and JC both conceptualized, drafted, revised, and approved manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AR and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


In the clinic, the diagnosis of Parkinson’s disease (PD) largely depends on clinicians’ experience. When the diagnosis is made, approximately 80% of dopaminergic cells in the substantia nigra (SN) have been lost. Additionally, it is rather challenging to differentiate PD from atypical parkinsonian disorders (APD). Clinially-available 3T conventional MRI contributes little to solve these problems. The pathologic alterations of parkinsonism show abnormal brain iron deposition, and therefore susceptibility-weighted imaging (SWI), which is sensitive to iron concentration, has been applied to find iron-related lesions for the diagnosis and differentiation of PD in recent decades. Until now, the majority of research has revealed that in SWI the signal intensity changes in deep brain nuclei, such as the SN, the putamen (PUT), the globus pallidus (GP), the thalamus (TH), the red nucleus (RN) and the caudate nucleus (CN), thereby raising the possibility of early diagnosis and differentiation. Furthermore, the signal changes in SN, PUT and TH sub-regions may settle the issues with higher accuracy. In this article, we review the brain iron deposition of PD, MSA-P and PSP in SWI in the hope of exhibiting a profile of SWI features in PD, MSA and PSP and its clinical values. Parkinson’s disease (PD) is characterized by resting tremor, rigidity, bradykinesia and postural instability accompanied by non-motor symptoms . The criThe most ideal biomarkers must be involved in the pathologic changes of PD and APD such that the potential biomarkers can indicate the underlying pathologic processes. The abnormal iron deposition in the deep brain nuclei in parkinsonism was first described by Lhermitte et al. in 1924 . Most stSWI exploits the differences in magnetic susceptibility of tissues, which describes the magnetic response of tissues placed in an external magnetic field, to develop an enhanced image contrast for conventional MRI –20. By aAccording to the reaction to the applied field, the substances can be classified into paramagnetic and diamagnetic. The paramagnetic substances mainly include ferritin containing ferric iron, deoxygenated hemoglobin, and ceruloplasmin, while the diamagnetic ones include myelin, calcium, and oxygenated hemoglobin . GeneralTo solve the problems of SWI, quantitative susceptibility mapping (QSM) is designed to quantify the iron content through measuring the susceptibility of certain areas directly , 39–43. The iron levels of basal ganglia are high , 45, 46.Even within the same deep brain nuclei, the iron distribution is uneven. Histological studies reveal that the SN pars reticulata (SNr) in the ventral SN is rich in iron, while the SNc in the dorsol SN is poor , 48. ZecA bunch of studies have approved that the speed of iron deposition across the brain is not even. In SN and GP, the iron concentration grows quickly in the early 20 years of life, then more gradually, and stops after 30 years of age , 49. ForIron functioning in the brain mainly exists in the forms of hemoglobin, iron-containing enzyme and non-haemin iron , 28, 45.Pathological and MRI studies indicate that the SN is the most relevant area of the brain in PD , 52–55. MSA and PSP are the most common APD and clinically the most important differential diagnosis for PD . AutopsyThe majority of research approved that SWI was feasible to indirectly quantify the iron content of different regions of brain through comparing the phase values in SWI which was highly correlated with the iron content , 22, 24.In recent years, a novel imaging biomarker called nigrosome 1, which is the sub-region of the SN, has been extensively studied by researchers. According to the immuno-staining of calbindin that can bind to calcium, the SNc is subdivided into nigrosome  and nigral matrix . Nigrosome 1 is the largest nigrosome containing the biggest group of dopaminergic cells and is affected in almost every PD patient . It was At present, the most promising signs in SWI that characterize MSA-P focus on signal changes in PUT due to its remarkably high iron level demonstrated by autopsy , 63, 76.Other deep nuclei, such as the CN and pulvinar TH (PT), were also studied. Meijer et al. reported that the iron content of the CN on the affected side of MSA-P was remarkably higher than that of PD in SWI . By contThere are different iron deposition patterns of PSP in SWI because of various forms of neuropathological processes . Meijer SWI is a promising biomarker that provides more information for early and differential diagnosis of parkinsonism, however present studies have several drawbacks that need to be addressed in the future. (1) Few research combined the SWI results of parkinsonism with pathological investigations , 71. (2)SWI characterizes brain iron deposition patterns of PD to illustrate the iron-related pathologic alterations in vivo and compensates for some drawbacks of routine MRI. Many researches have confirmed that SWI is a promising tool for the diagnosis and differential diagnosis of PD through discovering iron-related biomarkers, and more accessible in the clinic. Further studies should take the stages of neurodegenerative parkinsonism into consideration to acquire better correlations between SWI findings and neuropathologic results. With the underling pathological procedures illustrated by SWI, it will be possible to diagnose PD in the early stage and differentiate PD from APD.


Hypertrophic pulmonary osteoarthropathy (HPOA) is a paraneoplastic manifestation of gastric and, more frequently, lung carcinomas. It is characterized by extremity pain, clubbing, arthritis and periostitis of the long bones. Periostitis is the hallmark of HPOA and can be revealed with bone scintigraphy. Whole-body bone scintigraphy (WBBS) is very sensitive during the active lesion period and WBBS findings usually precede that of plain radiography. WBBS can also show improvement in the first 6 months following treatment, thus making it an important technique in the management and follow-up of these patients. While HPOA findings are usually seen in the lower extremities, involvement of both upper and lower extremities is a rare condition. In this case report, it is aimed to present findings of a 67-year-old male patient with lung cancer and complaint of extremity pain. We report on this patient to draw attention to HPOA of both upper and lower extremities.


Fluctuations in circulating estrogen and progesterone across the menstrual cycle lead to increased breast cancer susceptibility in women; however, the biological basis for this increased risk is not well understood. Estrogen and progesterone have important roles in normal mammary gland development, where they direct dynamic interactions among the hormonally regulated mammary epithelial, stromal, and immune cell compartments. The continuous fluctuations of estrogen and progesterone over a woman’s reproductive lifetime affect the turnover of mammary epithelium, stem cells, and the extracellular matrix, as well as regulate the phenotype and function of mammary stromal and immune cells, including macrophages and regulatory T cells. Collectively, these events may result in genome instability, increase the chance of random genetic mutations, dampen immune surveillance, and promote tolerance in the mammary gland, and thereby increase the risk of breast cancer initiation. This article reviews the current status of our understanding of the molecular and the cellular changes that occur in the mammary gland across the menstrual cycle and how continuous menstrual cycling may increase breast cancer susceptibility in women. The mammary gland is an essential reproductive organ, present in females of all mammalian species, which produces milk for both nourishment and immunological protection of newborns. It is a unique organ, as the vast majority of mammary gland development occurs postnatally, during puberty, pregnancy, and the postpartum period . MammaryA number of risk factors are associated with breast cancer, including family history of breast cancer, increasing age, and high breast density . There avia a complex network of intracellular and extracellular signaling pathways. It is widely accepted that mutations in mammary epithelial cells are the initial drivers of tumorigenesis. However, what is increasingly appreciated are the cell-to-cell interactions between epithelia and the surrounding stroma that affect DNA mutation rate, survival of DNA-mutated cells, and development of malignancy in the breast. Fluctuations in estrogen and progesterone across the menstrual cycle affect the abundance and function of mammary epithelial cells, stromal immune cells, and the ECM, and these changes are likely to be associated with biological mechanisms that cause increased breast cancer risk associated with menstrual cycling. The focus of this review is to collate current knowledge of the molecular and cellular events that occur during hormone-mediated menstrual cycling that affect epithelial and stromal cells in the mammary gland and how these contribute to increased breast cancer risk in women.The mammary gland consists of a number of different cell lineages including epithelial, hematopoietic, endothelial, and stromal . EpithelThe phases of the menstrual cycle in women are regulated by fluctuations in the pituitary gland hormones, follicle-stimulating hormone and luteinizing hormone, and the ovarian hormones, estrogen and progesterone . Hormonavia estrogen receptors (ERs)  , it alsors (ERs) . Importain situ breast carcinomas and atypical lesions are frequent in young and middle-aged women; they may remain in the non-invasive phase for 15–20 years before they develop into invasive breast cancers. In other words, hormone-regulated cellular events that occur during premenopausal years may induce persistent changes in the developmental fate of mammary epithelial cells. Alterations in signal transduction pathways, growth factors, and cell cycle regulators , WNT, CXCL12, and amphiregulin  to Th2 cytokines  by mammary epithelial cells . InteresOn the other hand, estradiol is shown to induce pro-inflammatory cytokine profile during the estrus phase in mice, an effect that was strongly mitigated by progesterone during other phases of the cycle . This inN-methyl-N-nitrosorea (NMU) between different phases of the ovarian cycle. Although there are some conflicting results in the literature, the majority reported that young rats exposed to chemical carcinogens at proestrus had a higher rate of mammary tumor incidence , which is a potent stimulatory factor in angiogenesis, doubled during the luteal phase (al phase . Angiogeal phase , 92. Moral phase , 94. TheUnderstanding the biological changes that occur over the course of the menstrual cycle could lead to the development of new approaches to prevent breast cancer in premenopausal women. Studies in rodent species, discussed above, suggest that susceptibility to initiating factors that lead to cancer might be elevated during specific stages of the cycle. Alcohol consumption increases breast cancer risk, potentially through increasing circulating estrogen and enhancing estrogen responsiveness, as well as increasing production of reactive oxygen species leading to DNA damage , 96. AnoA better understanding of how immune cell abundance and function fluctuates across the menstrual cycle may provide us with improved potential to harness the immune system to treat and prevent breast cancer. Immunotherapy and immunoprevention of cancer can involve immunization with a vaccine, passive transfer of tumor-specific antibodies, or adoptive transfer of immune cells that kill tumor cells. Changes in the types of macrophages in the breast, the cytokine microenvironment, and the phenotype of tumor-infiltrating lymphocytes across the cycle could affect the efficacy of both adaptive and humoral immune responses that recognize and eliminate tumorigenic cells , 100. FuIn addition to potential for breast cancer prevention and treatment, research on the effect of menstrual cycle stage on gene expression in the breast is critical in improving the utility of PCR-based diagnostic and prognostic tests for breast cancer. The relative expression of panels of genes, employed in tests such as Oncotype DX and Prosigna, classifies tumor subtype and predicts risk of disease recurrence, in order to guide treatment decision-making , 102. HoWomen who undergo early menarche and/or late menopause experience higher exposure to estrogen and progesterone, and a higher number of cyclical fluctuations of these hormone during their life time. Together with the direct effects of estrogen and progesterone on cancer initiation, there are a series of coordinated events during the menstrual cycle which are regulated by these ovarian hormones which have been implicated in increased breast cancer risk. Figure All the authors contributed intellectually to the content and writing of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.