We study theoretically the electronic transport and quantum fluctuations in single-molecule systems using thiophene rings as integrated elementary functions, as well as the dependence of these properties with the increase of the coupled rings, i.e., as a quantum wire. In order to analyze the current flow through these molecular systems, the thiophene rings are considered to be connected to metal contacts, which, in general terms, will be related to the application of voltages (bias voltages or gate voltages) to generate non-equilibrium behavior between the contacts. Due to the nonlinear behavior that is generated when said voltages are applied, it is possible to observe quantum fluctuations in the transport properties of these molecular wires. For the calculation of the transport properties, we applied a tight-binding approach using the Landauer–Büttiker formalism and the Fischer–Lee relationship, by means of a semi-analytic Green’s function method within a real-space renormalization (decimation procedure). Our results showed an excellent agreement with results using a tight-binding model with a minimal number of parameters reported so far for these molecular systems. The great technological revolutions that have been occurred over the years—such as the Industrial Revolution of the nineteenth century and the technological revolution that was based on the first transistor 1947), integrated circuits (1958) 947, inte,3, solidMolecular electronics can be defined as a technology that uses individual molecules or small groups of molecules to perform electronic functions . In the However, new assembly conditions arise, such as junctions formed with small molecules, strongly bonded to two electrodes. These conditions must be carefully considered because the mechanisms or connection devices (wires) must have certain conduction characteristics. This is the reason why much of the previous research has concentrated on the conductance characteristic of conjugated linear oligomers, taking into account properties as: (a) room temperature within a scheme of tunneling transport through the discrete electronic states of the molecule; (b) g others ,15,16,17N-thiophene rings taking into account the interactions between the molecule, rings, and contacts. In particular, the study includes molecular wires with one, two, and three thiophene rings placed between two metal contacts. Special attention is devoted to the electron transport by placing the system between metal leads and controlling the nonlinear bias voltage dependence of the current. Associated with such nonlinear character, quantum fluctuations in the transport properties—known as the noise power spectrum—can be observed. The noise power provides relevant information of the electronic correlation via the Fano factor In this work, the properties of quantum transport and current fluctuations are analyzed through molecular systems, specifically those systems that have in their molecular structure thiophene rings, which are known as molecular wires i.e., can be considered as solid-state structures of organic charge transfer (CT) with special electronic characteristics ranging from insulators to conductors ,10,11,12) limits ,19,20. W) limits ,22,23,24The manuscript is organized as follows: in In order to describe the single-molecule systems using thiophene rings as electronic devices, we have adopted the interaction of each system connected to the left and right contacts, as can bee seen in i atomic site of the thiophene ring and i atoms, where The full system is described by a tight-binding Hamiltonian, which is given by:i atomic site on the left and right sides of the molecular system.The Hamiltonian As we can observe, Equations –4) repr repr4) rp orbitals or In this new approach, the molecular states of the system (Left lead–thiophene molecule–Right lead) are expanded into a linear combination of atomic orbitals (LCAO) T(E)In order to obtain the Green’s functions as shown  can thenEquation  TF by calculating the NPCF (S) and the total current flowing through the aromatic molecules, with the following relationship [The noise power of current fluctuations (NPCF) is calculated by the expression ,25,26(1S=So∫−∞∞Ttionship :(13)F=S2For carriers ,28.The next section describes the analytical and numerical results obtained from the application of the decimation method and Landauer’s formalism for molecular systems with one, two, and three thiophene rings connected to terminal electrodes.The calculation of the transmission probability E, which compares the calculations made in this work with the results reported by Chang et al. [To validate the method used in this work, the reproduction of the reported calculations for the molecular systems with thiophene rings determined by a tight-binding numerical model with a minimal number of parameters. g et al. . The blaN resonant peaks, which represent the eigenvalues for each molecular system, where each ring is modeled as an atomic site (or quantum dot) with an energy site of The profile in the transmission probability shows N resonances, which must correspond to the molecular systems with N thiophene rings.We have taken the same energy values from Chang et al., where If we have a system with a thiophene ring, then there will be a resonant peak corresponding to the eigenvalue of its own state, which is at approximately The behavior of the profile in the transmission probability shows that as the number of thiophene rings increases, the width of the band also increases. This allows the position of the eigenstates of the system, particularly the state associated with the Highest Occupied Molecular Orbital (HOMO), to get close to the Fermi level. However, the amplitude of the transmission probability decreases as the number of rings increases. This behavior physically represents a decrease in the overlap between the charge density of the border states (sulfur states) and the thiophene rings.About the comparison between the present results and those from Chang et al., we have to stress that both models should arrive essentially to the same results. The theoretical model presented in  correspo profile  matches When our method has been validated, we then analyze the conductance, current, and shot noise through molecular wires composed of thiophene rings, considering the variation of the length of the molecular system and also the coupling between the contacts  v. C.01) , using tF), the shot noise goes from the Poisson limit (We find two limits of strong (red dots) and weak (black dots) couplings. In the limit of strong coupling, the amplitude of the current decreases as the number of thiophene rings in the molecular system increases. If we take the couplings between atoms of molecule ∼(0.1–6.0) eV, then the maximum amplitude of the current is on the order of microamperes , the gap of voltage decreases with the number of rings or length of the system. This means that, as the molecule–electrode coupling increases, the molecular system can pass from semiconductor  to a conductor system for a strong coupling, despite the fact that the amplitude of the threshold in the current decreases. The maximum amplitude of the current (saturation or threshold) of the systems happens for a voltage around 4.5 V and this amplitude then decreases with the increase of the thiophene rings, being 0.30 S) are calculated using the expression (S) that occur for an average value of the transmission probability and correspond to The quantum current fluctuations  is calculated using the expression  method and/or experimental results ,24,35.The renormalization method, which transforms the real space to an effective space, is suitable to study the transport properties in molecular systems, especially those that are large or have high complexity level of symmetry. As we could see, the idea of reducing a set of linear equations to nonlinear equations with energy and effective couplings  of the orbitals of the atoms at the end of the molecule with the Fermi level of leads. This behavior can be of interest because these kinds of sulphur systems  can be taken into account as possible prototypes for devices that can be used in molecular electronics, due to the strong coupling between S-lead, which causes a large location of charge in the interface. These molecular systems can be taken with and 4 eV ,37, valuThe above confirm the effectiveness of our method and thus gives us the feasibility of implementing it in future work to analyze transport properties in other molecular devices, or to calculate other thermal properties depending on wire length, properties of spin filter, among others, through molecular wire composed of thiophene rings.


Lower respiratory infections (LRIs) are the leading cause of death in children under the age of 5, despite the existence of vaccines against many of their aetiologies. Furthermore, more than half of these deaths occur in Africa. Geospatial models can provide highly detailed estimates of trends subnationally, at the level where implementation of health policies has the greatest impact. We used Bayesian geostatistical modelling to estimate LRI incidence, prevalence and mortality in children under 5 subnationally in Africa for 2000–2017, using surveys covering 1.46 million children and 9,215,000 cases of LRI. Our model reveals large within-country variation in both health burden and its change over time. While reductions in childhood morbidity and mortality due to LRI were estimated for almost every country, we expose a cluster of residual high risk across seven countries, which averages 5.5 LRI deaths per 1,000 children per year. The preventable nature of the vast majority of LRI deaths mandates focused health system efforts in specific locations with the highest burden. Geospatial modelling shows an overall decline in morbidity and mortality due to lower respiratory infections in Africa from 2000 to 2017, but also identifies subnational areas with residual high risk. These deaths are largely preventable; in particular, reducing risk factors such as household air pollution, stunting and no or partial breastfeeding have been shown to protect children against infection and death resulting from infection2. In addition, there are several direct interventions that can prevent or treat infections3, including vaccines to the predominant causes of LRI5, most notably Streptococcus pneumoniae, which is estimated to be responsible for 46.7% of LRIs across Africa6. Moreover, proven treatments such as antibiotics and supplemental oxygen can prevent death once an individual is infected7. Although childhood deaths due to LRIs are estimated to have fallen by more than 28.5% in Africa since 2000, this reduction is not geographically uniform, and LRIs still caused more than 432,000 deaths in children under 5 in 2017 (ref. 1). In response, the Global Action Plan for the Prevention and Control of Pneumonia and Diarrhoea (GAPPD) of the World Health Organization has called for the trichotomy of ‘protect, prevent and treat’8. The immediate need to assemble and analyse all available subnational data on the causes and drivers of LRIs was identified as key to empowering policy aimed at reducing LRI burden.Lower respiratory infections (LRIs) are estimated to account for 38.6% of infectious disease deaths and 14.9% of all deaths in African children8). Identifying areas or geographical clusters with the highest residual LRI burden is key to reducing the number of episodes and deaths due to LRI. Of more practical importance, identifying vulnerable populations that are either more likely to become infected by vaccine-preventable aetiologies or more likely to die once infected due to inadequate treatment can optimize targeted intervention strategies. Given the substantial subnational variation in other risk factors and causes of under-5 mortality10, as well as under-5 mortality itself11, country-level estimates of LRI burden probably mask these vulnerable populations. To fully understand the relative local drivers and causes of under-5 mortality, accurately capturing the covariation in risk factors and associated diseases is of paramount importance.In 2013, the GAPPD established the goal of reducing child mortality rates attributable to LRIs to below 3 in 1,000 persons and reducing severe LRI episodes to 75% of the 2010 values by 2025 , defined as the ratio of mortality to incidence, is an indicator of treatment-seeking practices and disease management. While some CFR interventions focus on the ‘protect’ component of the GAPPD guidelines, by improving a child’s overall health from birth, a number of more proximal interventions can greatly improve the prognosis of a child’s LRI disease in a clinic or hospital setting3. Although national estimates exist2, policy decisions and interventions rarely occur uniformly at the country level. Similarly, local variation in LRI incidence or mortality may be obscured by estimates restricted to national levels, hindering the ability to identify clusters or hotspots of LRI burden that cross administrative boundaries. While some risk factors for LRIs such as stunting14 and no or partial breastfeeding15 have been estimated subnationally, with considerable subnational heterogeneity, there are only a few analyses of spatial and spatiotemporal variation in LRI burden within selected regions in Africa18, and there has not been a comprehensive, subnational analysis of LRI burden for the continent as a whole.Precision public health focuses on identifying populations in need of specific interventions and providing those interventions in a timely manner19 to estimate posterior distributions of continuous continent-wide surfaces of LRI prevalence, incidence and mortality. We then use repeated draws from these posterior distributions and aggregate and summarize these to relevant administrative subdivisions  to identify those with a higher-than-average burden. We juxtapose subnational results with national-level estimates derived from the GBD 2017 to provide further context. Finally, we overlay these results onto detailed analyses of both the percentage of all LRI deaths attributable to S. pneumoniae and to LRI CFRs to identify regions of Africa most in need of improved vaccine coverage or case management, respectively.In this study, we present a systematic, comprehensive analysis of local variation in LRI morbidity and mortality in children under 5 across Africa between 2000 and 2017. We use Bayesian model-based geostatistics and an extensive geolocated dataset—describing 9,215,000 LRIs across 1.46 million children from 45,719 total data points, corresponding to 43,080 geolocated point-level survey clusters and 2,639 subnational, geolocated polygons—in combination with established methods from the Global Burden of Disease (GBD) 2017 studyhttp://ghdx.healthdata.org/record/ihme-data/africa-under-5-lri-incidence-prevalence-mortality-geospatial-estimates-2000-2017) and can be explored further via a custom data visualization tool .We conduct out-of-sample tests of model fit and model specification using fivefold cross-validation strategies. At the first administrative level, our estimates closely match the survey data . The greatest absolute decline in LRI rates among all divisions in Africa occurred in the Blue Nile state, Sudan, from 419.9 episodes (UI 383.0–458.0) per 1,000 in 2000 to 222.1 episodes (UI 62.1–508.5) per 1,000 in 2017  21.2–27.4) incident LRI episodes in children across all of Africa. Given the 56% increase in the population of children under the age of 5 in Africa, this is a moderate increase in LRI episodes from 20.6 million (UI 19.9–21.3) in 2000 Fig. . The ave017 Fig. . In 2017017 Fig. .Fig. 1LRIn spite of general declines in incidence across Africa, the GAPPD goal of decreasing severe incidence by 75% of its 2010 values by 2025 is ambitious. It is difficult to evaluate this goal subnationally, since severity data are scarce at the first administrative level. However, under the simplifying assumption that severe incidence is a constant proportion of all incidence across geographies, years, sexes and ages, extremely few divisions are on pace to hit the goal by the 2025 deadline  and the average LRI death rate was 4.9 per 1,000 (UI 4.6–5.1)  LRI deaths estimated among African children in 2017, an average mortality rate of 2.3 (UI 2.1–2.4) deaths per 1,000 Fig. 1. This w.1) Fig. . Among r.1) Fig. . Despite.1) Fig.  and 3. I.1) Fig. . The lar.1) Fig.  and 3.FiIn contrast to the severe incidence GAPPD goal, many divisions in Africa appear to have met the GAPPD mortality goal of <3 deaths per 1,000 children  were hotspots of mortality risk in 2000 Fig. . By 2017Unsurprisingly, countries that were large and consistent contributors to both the 2000 hotspots and residual hotspot in 2017 were those with the most modest relative reductions in mortality rate over the Millennium Development Goal period ; DRC, 52.2% (UI 34.6–64.3); Chad, 42.0%, (UI 20.7–57.8); Nigeria, 59.6%, (UI 46.0–69.7); and South Sudan, 31.9% (UI −1.6–54.3), as derived from the GBD 2017 data). While these reductions are substantial, they are behind the average reduction across Africa over this period (66.0% decrease (UI 51.2–76.7)).S. pneumoniae  deaths)1, but as with all-cause LRI mortality, the burden of LRIs attributable to S. pneumoniae has substantially fallen across almost all of Africa since 2000, with the notable exception of Zimbabwe, where S. pneumoniae mortality rates have increased by 40.3%  deaths per 1,000 . This programme is already estimated to be saving 4,400  children per year, but complete coverage (an unlikely scenario) could avert an additional 14,400  deaths per year, reducing the S. pneumoniae mortality rate in Nigeria by a further 18.8% (UI 11.4–25.2)  Fig. . To date.2) Fig. . Since iAs with overall LRI burden, a crucial step in targeting interventions is identifying where LRI CFRs remain highest. Across Africa in 2017, the CFR for childhood LRI was 18.3 (UI 16.3–19.8) per 1,000 and was estimated to be ≥30 per 1,000 in eight countries: Benin; Burkina Faso; CAR; Chad; Mali; Niger; Nigeria and South Sudan, compared to 18 countries in 2000 Fig.  and 2d. 1. We found that the rates of decline in incidence and mortality varied both between and within countries across first-level administrative divisions. Some countries had substantial and uniform reductions in LRI burden, while others performed less well in some areas. The divisions that struggled most were clustered into a single region spanning seven countries. Many of these divisions have had consistently high risks over the past 18 years, indicating that they have long been vulnerable. However, identifying the presence (and emergence) of this clustered region represents a unique opportunity for targeting interventions to those in most need. As demonstrated in southern Africa23, great successes are possible even in high-risk areas.Over the Millennium Development Goal period (1990–2015), dramatic declines at the national scale were documented across Africa, with most countries (42 of 52) reducing their under-5 LRI mortality rate by at least 40%Haemophilus influenzae type b vaccine and PCV coverage and lower exposure to indoor air pollution were the leading independent drivers of declining under-5 LRI mortality in southern sub-Saharan Africa2. In Botswana, for example, under-5 LRI deaths declined by an estimated 20.5% due to expanded access to PCV and by 11.0% due to H. influenzae type b vaccine coverage2. A more in-depth spatiotemporal analysis of the factors that lead to this and other successes could help identify important strategies for reducing both LRI incidence and CFR in the mapped high-risk areas.A recent country-level study found that expanded 22. By contrast, Nigeria currently has the highest CFR in Africa; thus, interventions that emphasize effective and timely treatment will have a large impact. Nigeria, like a few other countries in central sub-Saharan Africa, has both high CFRs and high incidence rates. As such, interventions focused on preventing infections (such as improved vaccine coverage) would likewise have substantial impact. The local distribution of risk factors—including indoor and outdoor air pollution, child growth failure, poor PCV and H. influenzae type b vaccine coverage and inadequate antibiotic use—could be combined with our morbidity and mortality estimates to create locally tailored intervention suites. Critically, in circumstances where increased antibiotic use is recommended, care must be taken to balance optimal intervention for the specific local conditions while simultaneously minimizing risk for an increase in antibiotic resistance24.The GAPPD framework to ‘protect, prevent and treat’ can guide intervention strategies depending on the local transmission setting. For example, Ethiopia has a below-average CFR, but partially due to its population size, it contains 3 of the top 10 divisions where the most children under 5 die from LRIs Figs.  and 3b. 25. Additionally, conversions from prevalence to incidence, estimates of the burden of S. pneumoniae, PCV coverage and the CFR analysis all leverage the GBD 2017 study modelled estimates, but GBD assumes that relationships and coverage values are constant within each country. While the conversion from incidence to mortality derived from the GBD study combines various data sources and allows for variation in CFR by country, year, sex and age, it does not currently incorporate the effects of comorbidities. Our geospatial approach borrows strength from neighbouring areas and may smooth over spatially or temporally focal epidemics. Furthermore, while the use of a continuous risk surface is a common approach for determining local risk, accounting for population movement and the clustered nature of populations in rural areas may improve local estimates. Our approach is focused on optimal prediction, not inference, and thus the fitted surfaces are not the optimal tool for assessing the impact of risk factors on risk. Regarding the post-hoc clustering analysis, these approaches, conducted on the modelled surfaces as opposed to the actual data, are used as indices of clustering as opposed to formal statistical tests. In addition, focused clustering analyses could also include more direct clustering approaches27, but given the coherent nature of the single clustered region identified, it is unlikely that those analyses would yield fundamentally different qualitative conclusions. Finally, there is strong evidence of age-related difference in risk within the 0–5 year age group, especially by aetiology29. Due to the nature of the data and methods we use, we are currently unable to parse mortality and morbidity estimates into finer age groups or split this burden by aetiology.These results are subject to several limitations. First, to produce continent-wide estimates, we make assumptions about data quality and consistency across a range of sources. For example, LRI prevalence surveys rely on self-reported data on cough and difficulty breathing and are subject to reporting and recall bias; we assume the same level of recall bias across all surveysThe burden of LRI in Africa has fallen dramatically since 2000, but this remarkable progress has not been universal across the continent. The estimates provided in this analysis illustrate the substantial local variability in childhood morbidity and mortality due to LRI, both across Africa and within individual countries. In particular, a single clustered region disproportionately contains much more risk than the rest of Africa. Exemplifying the local variability in risk, no entire country is contained within this region. Rather, subnational portions of countries may be identified as being part of the clustered residual hotspot. As Africa moves towards the 2025 GAPPD goals, those countries that are lagging behind will need specific, detailed focus to make satisfactory progress. The work presented in this study can be combined with targeted interventions to help children in the regions that are most neglected to reduce the mostly avertable burden of LRI.30. Prevalence, incidence and mortality were modelled on continuous continent-wide surfaces and were subsequently aggregated to the first-level administrative subdivisions (referred to as divisions). This study complies with the Guidelines for Accurate and Transparent Health Estimates Reporting recommendations .For this study, LRIs were defined as diseases of the lower airways including pneumonia and bronchiolitis. Severe LRI episodes, which we present in this study, are those requiring inpatient medical treatment as determined by a physician and based on World Health Organization Integrated Management of Childhood Illness guidelines2.We compiled 191 household surveys  from 2000 to 2017 with geocoded information from 45,719 total data points, 43,080 corresponding to survey clusters and 2,639 to subnational polygon boundaries. We included surveys that asked about 2- or 4-week prevalence of cough with difficulty breathing among children either under 3 or under 5 years old and allowed for geolocation below the country level. The prevalence of acute respiratory infection symptoms was adjusted to meet a standard case definition for LRIs (clinician-diagnosed pneumonia or bronchiolitis). The transformation of seasonally adjusted prevalence data from period to point estimates is described in detail in the GBD 2016 study of global, regional and national LRI mortality and morbidityWe differentially scaled our continuous continent-wide surface estimates of the spatial pattern of prevalence to national-level estimates of LRI prevalence and incidence from the GBD 2017 study by calculating a population-weighted mean such that the national estimates presented in this study match those from the GBD 2017 study. To produce the severe incidence estimates, we multiplied our prevalence estimates by the GBD fraction of LRI episodes categorized as severe divided by average duration. CFRs by country and year were obtained from the GBD by dividing the country-specific mortality of LRI in a given year by the country-specific total incidence in that year. Finally, we estimated our continuous continent-wide surface of mortality by multiplying our incidence surface by the appropriate country-year CFRs. Detailed data processing descriptions are provided in Section 2.0 of the 19. Briefly, this framework uses a spatially and temporally explicit hierarchical logistic regression model to predict LRI prevalence in locations with sparse observations where points that are closer together in space and time—and which have similar covariate patterns—are expected to have similar LRI prevalence. Because the prevalence of LRIs has been shown to be influenced by social, structural and environmental factors32, we selected a set of 12 continuous geographical covariates based both on previous mapping efforts and past GBD analyses of LRI for inclusion in the model . Uncertainty was calculated by taking 1,000 draws from the posterior joint distribution of the model; each point value is reported with an uncertainty interval that represents the 2.5th and 97.5th percentiles of those 1,000 draws. Additional methodological details can be found in the Adjusted point prevalence data were used as inputs to a Bayesian model-based geostatistical framework  or lower (coldspot) than the expected value as informed by the total set36. We then conducted a sensitivity analysis to determine the impact of the spatial and temporal correlations structures inherent to the Bayesian geostatistical approach on the clustering hotspot analysis. Moreover, we conducted the same analysis on each draw from the posterior distribution of mortality risk and summarized the frequency for which each first administrative division was identified as a ‘hotspot’. This sensitivity analysis avoids the use of the standard reference distribution for the local statistic and as such is adjusting for the spatial variation in the background population size. The second method identified clusters of higher-than-average burden arithmetically by identifying all divisions with the highest estimated mortality rate that account for a fixed percentage of all deaths and then counting the number of contiguous divisions identified. It is important to note that for both analyses, ‘hotspot’ is a relative term, comparing the risk in one area to other areas within a single year. Overall burden has decreased from 2000 to 2017, so locations that are identified as hotspots in 2017 would not have qualified in 2000. To emphasize this, when identifying clusters of remaining high risk in 2017, we refer to them as residual hotspots.We used two post-hoc approaches to identify regions or clusters of higher-than-average burden from our estimates: the Getis–Ord local G statisticFurther information on research design is available in the Supplementary InformationSupplementary Guidelines, Supplementary Results, Supplementary Figs. 1–20, Supplementary Tables 1–13 and Supplementary References.Reporting Summary


Euglena gracilis, which can be processed to produce linear nanofibers capable of interacting with specific receptors present on cell membranes. The effects of these nanofibers, already investigated on plants, animals, and humans, will be analyzed.β-glucans, heterogeneous glucose polymers present in many organisms, have the capability to activate the innate immune system. Efficacy of activation depends on purity of the compound, molecular structure, polymerization degree, and source. One of the purest forms of crystallized β-(1–3)-glucan present in nature is the paramylon extracted from the WZSL non-chloroplastic mutant of  All these features influence their physiological functions and biological activity. These polysaccharides commonly consist of a main chain of β-1,3) and/or β-1,4)-glucopyranosyl units in non-repeating non-random order, with side chains of varying lengths ,3 and/or,3. β-glu-glucopyrDectin-1 is indicated as the preferential receptor for linear β--linked glucans ,13,14,15Euglena gracilis can be considered highly pure since it lacks any contamination from cellular components . Such purity is confirmed by optical and scanning electron microscope examination and by NMR spectrum, which indicates the presence of 100% glucose , th, th27], Saccharomyces were not absorbed to the cell surface of PBMC due to their sizes, which did not fit with Dectine-1 receptors. Though both these preparations were internalized, this phagocytic activity did not lead to any activation of the immune system, i.e., no dendritic cells differentiated [This signaling cascade guarantees a safe activation of the innate immune system, as demonstrated by the presence of newly differentiated dendritic cells . The paramylon administration continued over the entire experimental period (2 + 4 weeks). Since the progression of fibrosis is mainly due to inflammation and collagen accumulation, we evaluated liver damage markers and collagen content (Picrosirius red staining and hydroxyproline determination).4-induced loss of weight and prevented the increase of aspartate aminotransferase (AST) typical of hepatocyte damage. It also reduced the hyper-echoic areas caused by the toxic treatment, restoring the normal tissue consistency and appearance. CCl4-treated liver had a trabecular ‘fishnet’ texture typical of mild to severe edema, while normal color and consistency was restored in paramylon treated livers, indicating a relieved hepatic injury  is a major environmental factor limiting photosynthesis and contributing to the progressive salinization of world arable land, consequently reducing crop production all over the world , and tomSolanum lycopersicum cultivar Micro-Tom, chosen because of its short life cycle (≤3 months) and small size, were challenged with drought stress with or without root treatment with paramylon nanofibers [Paramylon nanofibers applied to the root system of tomato plants have already shown to modulate hormone levels, photosynthesis, and water use efficiency . To verinofibers . Plants Ecophysiological, physical chemical and quality parameters were monitored throughout the life cycle of tomatoes and compared with those of well-watered, untreated plants. Flowering and fruit ripening of stressed paramylon treated plants were about 2 weeks precocious respect to untreated well-watered plants, whereas fruits of stressed untreated plants do not ripe beyond category II .−1 day−1 can be lowered down to 0.36 L plant−1 day−1. Ecophysiological parameters, , were dramatically influenced by water stress, all of them undergoing a continuous decrease to saturation. Root treatment with paramylon nanofibers allowed all the parameters to recover to the values of well-watered plants. These results indicate an effective action of paramylon nanofibers on stomatal behavior, whose control improves water use efficiency, hence prevents dehydration. This action is associated with a transient modification of the content of main plant hormones, i.e., abscissic acid, jasmonic acid and salicylic acid [Thanks to paramylon action, we observed that the optimal water regimen of about 8.64 L plantlic acid .The great increase of physical-chemical and quality parameters such as the antioxidant compounds  ,37,38,39Moreover, the higher dry matter content  allows a better post-harvest storage capability extending the commercial period and increasing the commercial product value. These results confirm the involvement of paramylon nanofibers as elicitors of plant response to abiotic stress and their bio-stimulant activity in increasing plant adaptation capacity to environmental stimuli.S. cerevisiae particles are in the range of pathogenic bacteria (2–3 µm); still, they are not capable to interact with the Dectin-1 receptors present on the surface of immune system cells. Phagocytosis of these two preparations does not lead to any activations of the immune system since no dendritic cells differentiate [2 diffusion from air, which eventually leads to an improvement of water use efficiency. Activation is induced only by paramylon nanofibers because they consist of a moiety of 9- or 10-mer of glucose, the minimum β-glucan recognized by Dectin-1 receptor [Both paramylon granules and nanofibers obtained by processing the granules have been tested in our research, and nanofibers have shown the capability to effectively interact with PRRs eliciting responses in animal and plant models. Our results indicate that even though the dimensions of paramylon granules and rentiate ,24. Besireceptor ,13,28.Euglena mutant can be considered an effective and desirable producer of paramylon due to its ability to convert and conserve glucose  in the


To provide adequate conditions for the regeneration of damaged bone, it is necessary to develop piezoelectric porous membranes with antioxidant and anti-inflammatory activities. In this study, composite nanofibers comprising poly(vinylidene fluoride) (PVDF) and a polyhedral oligomeric silsesquioxane–epigallocatechin gallate (POSS–EGCG) conjugate were fabricated by electrospinning methods. The resulting composite nanofibers showed three-dimensionally interconnected porous structures. Their average diameters, ranging from 936 ± 223 nm to 1094 ± 394 nm, were hardly affected by the addition of the POSS–EGCG conjugate. On the other hand, the piezoelectric β-phase increased significantly from 77.4% to 88.1% after adding the POSS–EGCG conjugate. The mechanical strength of the composite nanofibers was ameliorated by the addition of the POSS–EGCG conjugate. The results of in vitro bioactivity tests exhibited that the proliferation and differentiation of osteoblasts (MC3T3-E1) on the nanofibers increased with the content of POSS–EGCG conjugate because of the improved piezoelectricity and antioxidant and anti-inflammatory properties of the nanofibers. All results could suggest that the PVDF composite nanofibers were effective for guided bone regeneration. A variety of techniques have been tried for the regeneration of bone defects, including bone grafting, distraction osteogenesis, and guided bone regeneration (GBR) ,2,3. AmoNanofibrous membranes have structural similarity to the extracellular matrix (ECM) and high specific surface area . GBR memPoly(vinylidene fluoride) (PVDF) is an attractive semicrystalline polymers for use in biomedical applications because of its efficient piezoelectricity and biocompatibility ,14. The Polyhedral oligomeric silsesquioxane (POSS) molecules have a unique nanostructure comprised of an inorganic silsesquioxane cage core and an organic functional group shell . A variePiezoelectric materials can change their surface charge without external energy power source . SeveralwM = 275000 g/mol), EGCG, lipopolysaccharide (LPS), horseradish peroxidase (HRP), hydrogen peroxide , dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), tetrahydrofuran (THF), and isopropyl alcohol (IPA) were obtained from Sigma-Aldrich Co. . Xanthine oxidase , xanthine, and nitro blue tetrazolium (NBT) were purchased from Wako Pure Chemical Industries . Aminopropylisobutyl polyhedral oligomeric silsesquioxane (API-POSS) was purchased from Hybrid Plastics . Osteoblast-like cell line MC3T3-E1 derived from mouse calvaria was acquired from the American Type Culture Collection  and a murine macrophage cell line RAW 264.7 was purchased from the Korean Cell Line Bank . All materials were used as received without further purification.PVDF . The structures of EGCG, API-POSS, and the POSS–EGCG conjugate were determined by 1H NMR spectroscopy .The POSS–EGCG conjugate was synthesized as follows. API-POSS  and EGCG  were first dissolved in 30 mL of a mixed solvent of THF/IPA/deionized water (DW) at a ratio of 2:1:1 before adding a solution of HRP  dissolved in 3 mL of Dulbecco’s phosphate-buffered saline . To this resultant solution, 5 mL of 5 wt% H1H NMR of EGCG , δ (ppm) = 6.91 (CH of D-ring), 6.52 (CH of B-ring), 6.10, 6.07 (CH of A-ring), 5.48, 4.97 (CH of C-ring), 2.88 (CH2 of C-ring).1H NMR of API-POSS , δ (ppm) = 2.83 (CH2NH), 1.84 (CHCH3), 1.58 (CH2CH2CH2), 0.95 (CHCH3), 0.56 (SiCH2).1H NMR of POSS–EGCG , δ (ppm) = 6.81 (CH of D-ring), 6.41 (CH of B-ring), 5.95, 5.84 (CH of A-ring), 5.36, 4.98 (CH of C-ring), 2.77 (CH2 of C-ring) for EGCG, 2.69 (CH2NH), 1.82 (CHCH3), 1.58 (CH2CH2CH2), 0.95 (CHCH3), 0.54 (SiCH2) for API-POSS.Before electrospinning the PVDF composite nanofibers, the PVDF (1.8 g) was dissolved in 10 mL of a mixed solvent of DMF/THF in a ratio of 1:1 to obtain an 18 wt% solution. After stirring 12 h, the POSS–EGCG conjugate was added to the PVDF solution with a different weight ratio at 60 °C for 8 h. The contents of POSS–EGCG conjugate in solutions were 0, 2, 4, and 6 wt% with respect to the weight of the PVDF. The mixed solution of PVDF and the POSS–EGCG conjugate was filled into a 20 mL standard syringe with a 22 G stainless steel needle . Then, this solution was electrospun into nanofibers at room temperature with a feeding rate of 2 mL/h feed rate, a voltage of 18 kV, and a working distance of 15 cm. After electrospinning, the PVDF composite nanofibers were peeled off from the stainless steel plate and vacuum dried for 12 h.The morphology of the PVDF composite nanofibers was visualized by scanning electron microscopy  at an acceleration voltage of 5 kV after sputter coating of samples with gold. Image-Pro Plus software  was used to determine the average fiber diameters for the composite nanofibers. Energy dispersive spectroscopy (EDS) spectra were obtained for the determination of the distribution profile of the POSS–EGCG conjugate on the nanofiber surface.−1. Attenuated total reflectance (ATR) mode was used to analysis the characteristic bands over a range from 400 to 1600 cm−1. The relative β-phase content (F(β)) in the samples was determined from the following equation [α and Aβ represent the absorbance of peaks related to the α- and β-phases at 760 and 840 cm−1, respectively, and Kα and Kβ are the absorption coefficients at the respective wavenumbers, which are 7.7 × 104 cm2/mol and 6.1 × 104 cm2/mol.Fourier transform infrared spectroscopy (FTIR) measurement was performed using an ALPHA spectrometer  with a resolution of 4 cmequation .F(β)=AβX-ray diffraction (XRD) patterns were also used to evaluate the crystalline phases of the PVDF composite nanofibers and thus XRD measurements were performed using a Rigaku D/MAX-2500V/PC X-ray diffractometer (Japan) with high intensity Cu Kα radiation at 40 kV/100 mA. The diffractograms were scanned in a 2θ range from 10° to 50°. The composite nanofiber samples were analyzed using X-ray photoelectron spectroscopy (XPS) to obtain the surface elemental composition. XPS measurements were performed using a Quantera SXM  equipped with a monochromatic Al Kα X-ray source (1486 eV). The photoelectron take-off angle was fixed at 45° relative to the sample surface. Besides XPS measurements, static contact angles were measured with a DSA 100 contact angle meter  to examine the hydrophobicity of the composite nanofiber. Each contact angle of the sample is an average of five measurements. The mechanical properties of the composite nanofibers were investigated with a TO-101 universal testing machine  with 2 kN load capacity at a rate of 10 mm/min. All samples were cut into rectangular specimens with a size of 3 cm × 1 cm and tested five parallel measurements.2. Before assaying, the composite nanofibers were cut into circular shape with a diameter of 15 mm, followed by putting into 24-well tissue culture plate and fixing with glass ring (inner diameter = 11 mm). Then, they were sterilized with 70% ethanol and rinsed with DPBS and α-MEM. After drying, the composite nanofibers were once again sterilized under UV radiation for 3 h.MC3T3-E1 cells were used to evaluate the in vitro bioactivity of the PVDF composite nanofibers by a cell proliferation assay in alpha minimum essential medium (α-MEM) containing 10% of fetal bovine serum (FBS) and 1% of penicillin–streptomycin at 37 °C with 5% CO4 cells/well) was placed onto the sterilized samples and cells were cultured for different periods of time. After culturing for 1, 3, 5, 7, and 14 days, culture media were replaced with 0.2 mL of the MTT solution and further incubation of the cells was maintained for another 4 h. Next, we removed the remaining media and added 1 mL of DMSO to solubilize the precipitated formazan crystals. Finally, the resulting supernatant was transferred to 96-well plate with 0.2 mL per well. The absorbance was determined at 570 nm using a spectrophotometric plate reader . Furthermore, after days 3 and 7 of cell culture, the cell-nanofibers were fixed in 4% glutaraldehyde for 1 h, and then dehydrated using different concentrations of ethanol , followed by vacuum-drying. The morphologies of dried samples were observed using SEM after sputter coating with gold.The proliferation of viable cells on the composite nanofibers was determined using the 3--2,5-diphenyltetrazoliumbromide (MTT) assay. One milliliter of cell suspension  at periods of 3, 5, 7, and 14 days after cell seeding. The composite nanofibers seeded with cells washed with DPBS and incubated in 0.5 mL of DW containing 0.2% Triton X-100. The cell lysates were mixed with 4 cells.Bone mineralization capability of the composite nanofibers was assayed by the alizarin red S (ARS) staining method. ARS can selectively bind to calcium ions and thus calcium deposition is easily measured by the use of ARS. After 3, 5, 7, and 14 days of cell culture on the different nanofibers, the cells were fixed with 4% glutaraldehyde for 30 min and then stained with 1 mL of 40 mM ARS (pH 4.1). After incubation for 20 min, the specimens were washed thrice with DW to remove unreacted ARS and dissolved in 1 mL of 10% cetylpyridinium chloride. The absorbance of the supernatant at 540 nm was obtained using microplate reader and was normalized to 1 × 104 cells/well) were precultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% heat-inactivated FBS, 100 U/mL of penicillin, and 100 μg/mL of streptomycin for 24 h at 37 °C before adding 1 μg/mL of LPS into the cell culture plate. After activation of the cells for 24 h, the amount of IL-6 released into the media was measured by mouse IL-6 enzyme-linked immunosorbent assay kit  according to the manufacturer’s protocol.The effect of the PVDF composite nanofibers on the expression of the inflammatory cytokine interleukin-6 (IL-6) was measured using the RAW 264.7 cells. For the quantification of IL-6 production, the cells . A significant difference was defined for values of 1H NMR spectroscopy ) in the PVDF composite nanofibers. As presented in  of PVDF ,28. MoreThe surface elemental composition of the PVDF composite nanofibers was assessed using XPS. The XPS survey scan spectrum of the pure PVDF nanofiber exhibited two separate peaks which corresponded to the F 1s orbital (688 eV) and the C 1s orbital 287 eV) a. After 7 eV a. AThe investigation on the surface hydrophilicity of biomedical membranes is very important to confirm the affinity between and the membranes and the cell. Suitable hydrophilicity can influence the attachment and proliferation of the cells on the surface of the membranes. Therefore, the performance of membranes in biomedical applications may depend on the hydrophilicity . Here weGBR membrane should be strong enough to withstand the force during new bone tissue regeneration. Thus, the effect of the addition of the POSS–EGCG conjugate on the mechanical properties of the composite nanofibers was assayed. As shown in The in vitro bioactivity of the PVDF composite nanofibers was evaluated to assess the potential of the nanofibers as GBR membranes. The cell proliferation on the PVDF composite nanofibers was studied by the MTT assay, resulting that the relative cell viability of the MC3T3-E1 cells cultured on the composite nanofibers indicated good cytocompatibility of the nanofibers . For allDuring tissue regeneration, piezoelectricity can induce an increase in the surface charge density of ECM materials for the delivery of an electrical stimulus without external energy sources and consequently causes higher levels of cell proliferation and differentiation ,16. In aTo confirm cell growth on the PVDF composite nanofibers, the interactions between the cells and nanofibers was observed using SEM on periods of 3 and 7 days after cell seeding. SEM images exhibited that the cells adhered and spread on the surface of the composite nanofibers, and cell proliferation increased time-dependently in all the tested nanofibers . This inALP catalyzes the cleavage of organic phosphate esters and plays an important role during the formation process of bone nodule ,2. ALP aThe mineralized bone nodules are usually formed via three main stages such as cell proliferation, ECM development, and mineralization. Therefore, the osteogenesis capability of the cells as a late-stage marker for osteogenic differentiation can be estimated by the determination of bone-like calcium deposits produced on different PVDF composite nanofibers using ARS staining ,4. AfterOsteoblasts can produce bone-active cytokines such as tumor necrosis factor-α and IL-6 ,38. In bIt was found that PVDF and PO06 promoted the expression of the IL-6 markedly due to the contamination by LPS and subsequent macrophage activation . HoweverIn this study, we fabricated PVDF composite nanofibers as potential membranes for bone tissue regeneration using an electrospinning process. The composite nanofibers obtained had fully interconnected porous structures adequate to promote the cell proliferation and the efficient transport of nutrients and metabolic waste. The content of POSS–EGCG conjugate present affected the piezoelectric, antioxidant, and anti-inflammatory properties of the nanofibers, which finally influenced the in vitro bioactivity of the PVDF composite nanofibers. PE06 facilitated the proliferation and differentiation of the MC3T3-E1 cells more effectively compared to the other nanofibers. Based on these results, our simple method is very useful to prepare PVDF composite nanofibers which may contribute to the development of new scaffolding materials for bone tissue regeneration.


Antipsychotic treatment resistance in schizophrenia remains a major issue in psychiatry. Nearly 30% of patients with schizophrenia do not respond to antipsychotic treatment, yet the underlying neurobiological causes are unknown. All effective antipsychotic medications are thought to achieve their efficacy by targeting the dopaminergic system. Here we review early literature describing the fundamental mechanisms of antipsychotic drug efficacy, highlighting mechanistic concepts that have persisted over time. We then reconsider the original framework for understanding antipsychotic efficacy in light of recent advances in our scientific understanding of the dopaminergic effects of antipsychotics. Based on these new insights, we describe a role for the dopamine transporter in the genesis of both antipsychotic therapeutic response and primary resistance. We believe that this discussion will help delineate the dopaminergic nature of antipsychotic treatment-resistant schizophrenia. D2 rere used , 73, 74.nds used . This inmarkedly , 77. Theefficacy .2 receptors is not a working antipsychotic mechanism for a significant number of patients with schizophrenia  of a proportion of Dophrenia . In factccupancy . Clozapiccupancy , 78, 79,ccupancy , 32, 80.kinetics , we begikinetics , 38. Somkinetics , which ckinetics . Diminisccupancy . These dccupancy –85.2 receptor upregulation, consequent to chronic D2 receptor blockade  could involve antipsychotic-induced dopamine receptor supersensitivity, potentially resulting from Dblockade , 86, 87.blockade –90. In lblockade , 91, 92 blockade , 93–95. h groups , 97. Rec imaging  or ex viiography , 91. Ant density . Thus, csitivity , 98, 99,eceptors .2 receptor density, chronic antipsychotic treatment can also increase D2 receptor function, and this has been linked to dopamine supersensitivity and acquired antipsychotic tolerance. When D2 receptors are coupled to Gi/o proteins, they are in a functional, high affinity state for dopamine (referred to as D2HIGH). When D2 receptors are uncoupled to Gi/o proteins, they are in a functionally inert, low affinity state for dopamine (D2LOW). As such, the proportion of D2HIGH can modulate dopamine signaling via D2 receptors. The link between antipsychotic tolerance and changes in striatal D2HIGH sites comes largely from work in animal models showing that chronic antipsychotic treatment increases striatal D2HIGH levels , which are postsynaptic to dopaminergic terminals projecting from the midbrain, among other regions; for an overview, see Refs.  and the short (D2S) forms of the protein  relative to D2S mRNA  appear to selectively bind those receptors expressed in the striatum (a structure with high D2L/D2S ratio)  appear to confirm an antipsychotic selectivity for D2L (2 receptor occupancy in the striatum than in the midbrain (SN).  , 123, sued cells –127, sat for D2L –131. Conin (SN). , 133.2L mRNA in the caudate nucleus  and in postsynaptic neurons (heteroreceptors). Dopamine stimulation of D2 autoreceptors at terminals decreases synaptic dopamine release, while stimulation of somatic D2 autoreceptors instead decreases the firing activity of these cells  during depolarization and accelerating the decay rate of inactivation tropane)  , 188. Hotropane) , 189, aleceptors –192. Acceceptors . In suppifensine –196. Thiifensine , 188. Siifensine , 197 andifensine , then DAifensine –202 as wex vivo studies support a direct interaction between antipsychotics and the DAT. Under normal physiological conditions, increased dopamine release rapidly upregulates DAT membrane expression (in vivo (ND) during antipsychotic failure, suggesting the putative relevance of dopamine clearance for achieving antipsychotic therapeutic response, at least in animal models. Interestingly, changes affecting DAT expression and corresponding behavioral responses to antipsychotics are accompanied by a stable and clinically relevant D2 receptor blockade (69%) and by increased or decreased extracellular dopamine in the striatum, during the expression of antipsychotic efficacy and failure, respectively . During phasic dopamine release , dopamine reaches presynaptic autoreceptors, producing antipsychotic-dependent dopamine-mediated autoinhibition and a corresponding antipsychotic efficacy  of autoreceptors may prevent the occurrence (or reduce the likelihood) of autoinhibition altogether Figure 4achinery –163, 172nt users  and are nt users .It should be noted that the mechanisms described here and depicted in via drug-induced downregulation of DAT expression. This individual may seek medical intervention upon first experience of psychosis, at which time he or she will receive antipsychotic treatment and may already face reduced therapeutic efficacy due to the drug-related changes in DAT expression. On the other hand, if patients have no history of addictive substance consumption before starting antipsychotic treatment and begin using moderate doses of addictive drugs thereafter, we speculate that the effects of antipsychotics and certain categories of addictive substances on the expression and function of the dopaminergic machinery  may counterbalance one another  and by the Brain & Behavior Research Foundation (NARSAD Young Investigator Award 2018). AK is supported by the National Institutes of Health (DA044782). A-NS is supported by a salary award from the Fonds de la Recherche du Québec-Santé (28988).A-NS is a consultant for H. Lundbeck A/S. This had no influence on the manuscript’s content. The remaining 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.


Gayralia oxysperma (Go3) to inhibit the effects of venom from Bothrops jararaca and Lachesis muta. B. jararaca or L. muta venoms were incubated together with sulfated heterorhamnans from Go3, and the in vitro  and in vivo  activities of venoms were assessed. Additionally, Go3 was injected before and after the injection of venoms, and the toxic activities were further tested. When incubated with the venoms, Go3 inhibited all activities, though results varied with different potencies. Moreover, Go3 neutralized hemorrhagic, myotoxic, and edematogenic activities when injected before or after injection with B. jararaca and L. muta venom. Go3 also blocked the coagulation of plasma in mice caused by the venoms in an ex vivo test. Therefore, Go3 has the potential to be used as antivenom for B. jararaca and L. muta bites, notably exhibiting higher efficacy on L. muta venom.Worldwide, snakebites have serious implications for human health. The administration of antivenom is the official treatment used to reverse the toxic activities of envenomation. However, this therapy is not efficient to treat the local effects, leading to the amputation or deformity of affected limbs. As such, alternative treatments are needed. Here, we analyze the ability of a polysaccharide from the green marine alga  Crotalus, Bothrops, and Lachesis. The latter two genera are responsible for the highest incidence (90%) and lethality (1%), respectively [Snakebites represent a global public health problem. According to the World Health Organization (WHO), snakebites are a neglected tropical disease that affects 5.5 million people annually, resulting in approximately 400,000 amputations and 120,000 deaths. However, epidemiological studies are inadequate and unreliable, resulting in such figures being underestimated . In BrazTo date, apart from supportive care, the official and effective treatment for envenomation by snakebite is the intravenous administration of antivenom (immunoglobulins) . In mostMonostroma, Ulva, and Enteromorpha are known to display a variety of biological activities, such as antiviral [Gayralia (Chlorophyta) comprises two species: G. oxysperma and G. brasiliensis [Gayralia genus, such as the monostromatic Monostroma sp., are cultivated and used in the food and cosmetics industries [Gayralia oxysperma is composed of sulfated heterorhamnans [G. oxysperma (Go3) was obtained by aqueous extraction at 80 °C , and presented a composition of 49.6% total carbohydrates and 17.0% uronic acids. Go3 is highly sulfated (25.3%), and presents rhamnose as the major monosaccharide, as well as minor amounts of xylose, glucose, galactose, glucuronic acid, galacturonic acid, and very low percentages of arabinose and mannose . Chemical and spectroscopic analyses performed with the major sulfated heterorhamnan constituent of Go3 (70%) demonstrated that 3-linked units are sulfated on C-2, C-4, disulfated, and unsulfated in a molar ratio of 1:077:038:0.46, respectively. The 2-linked rhamnosyl units are principally sulfated on C-4, sulfated on both C-3 and C-4, and unsulfated . Furthermore, glucuronic 2-sulfate and galacturonic acids and xylosyl units are components of the side chains of this partially-branched heterorhamnan. The heterorhamnans isolated from G. oxysperma exhibited potent antiviral activity against the herpes simplex virus (HSV-1 and HSV-2), and were devoid of cytotoxic effects at concentrations up to 1000 µg mL−1 when assayed on Vero cells [G. oxysperma and their products  increased as they became cytotoxic against tumoral cells (U87MG). The inhibitory effect on human glioblastoma cells was correlated with the molecular weight and sulfate location of the partially depolymerized products [Palisada flagellifera and Laurencia aldingensis, respectively, were able to inhibit the toxic effects of Lachesis muta venom [Fucus vesiculosus inhibited the myotoxic activity of some crotaline snake venoms; in these cases, the formation of a complex between fucoidan and isolated myotoxins has been postulated as the mechanism of action for this natural polysaccharide [Among marine organisms, green algae have been extensively investigated due to their bioactive compounds, widespread distribution, and large biomass production. Sulfated polysaccharides obtained from the genera ntiviral ,17,18,19ntiviral , antihypntiviral ,22, and ntiviral . Monostriliensis . Some spdustries . These sdustries ,24,25,26products . Indeed,ta venom ,29. A fuccharide . Surprisccharide .G. oxysperma (Go3) sulfated heterorhamnans against some toxic activities of L. muta and B. jararaca venoms.Therefore, we evaluated the effect of L. muta or B. jararaca (2–40 μg mL−1) venom induced hemolysis, proteolysis, or coagulation in a concentration-dependent manner. One minimum indirect hemolytic concentration , effective concentration , or minimum coagulation dose  of venoms was incubated with Go3 , and then assays were performed. As shown in w/w), respectively. Go3 inhibited B. jararaca proteolysis more efficiently than that of L. muta, since 100% of B. jararaca proteolysis was inhibited at 1:20 venom:Go3 ratio, whereas for L. muta venom, a 60% inhibition was achieved (−1) did not cause hemolysis, proteolysis, or coagulation (data not shown).achieved B. In conachieved C. Go3 alL. muta venom was evaluated using an ex vivo methodology  and activated partial thromboplastin time (aPTT) coagulation tests were performed. Moreover, another experimental approach was evaluated in which L. muta venom was injected s.c., and then followed 30 min later by an i.v. injection of Go3. After 90 min, blood was collected and coagulation tests were performed. As shown in L. muta venom, while the treatment of mice with Go3 restored plasma coagulation to normal levels in PT and aPTT tests. Injection of only Go3 did not alter plasma coagulation compared to mice that received a saline injection only.The effect of Go3 on plasma coagulation of hodology . Mice reL. muta (24 µg/mouse) or B. jararaca (30 µg/mouse) venom induced a hemorrhage halo of 20 mm, which corresponds to two minimum hemorrhagic doses (MHDs), and such halos were considered to be 100% of hemorrhagic activity. When mixed with the venoms, Go3 (140 µg/mouse) inhibited 70% and 40% of the hemorrhagic activity of L. muta and B. jararaca venoms, respectively (L. muta (black columns) or B. jararaca (white columns) venoms were injected s.c. before  was injected s.c., and a single injection (1 injection) or two injections (2 injections) of Go3 were given 1 h later  and then injected s.c. into mice, and then the edema was analyzed using the aforementioned procedure  mixed with saline died at 110 min (B. jararaca venom was incubated with Go3 (140 μg/mouse) followed by i.p. injection of the mixture into mice, deaths occurred at 350 min. Go3 also delayed mice from death even when the Go3 injection was performed 15 or 30 min before (prevention protocol) or after (treatment protocol) injection of B. jararaca venom, and this neutralization efficacy was quite similar in both protocols  from mice plasma. Injection i.m. of L. muta venom (15 μg/mouse) mixed with saline (S) produced a release of CK of the muscle of mice at of 950 U L−1 (L. muta venom was mixed with Go3 (140 μg/mouse), the myotoxic activity of the venom was fully inhibited, since no trace of CK was detected  was injected 15 min later at the same site of the venom injection, with myotoxicity being evaluated thereafter. As seen in L. muta venom-induced myotoxicity through the treatment protocol. The injection of Go3 or saline only did not induce the release of CK of plasma in mice.Myotoxic activity was not performed for The incidence and mortality indices of snakebites are higher than other neglected diseases, such as dengue, cholera, Chagas disease, leishmaniasis, yellow fever, and schistosomiasis ,2. Due tB. jararaca and L. muta venoms. This is in contrast to both Brazilian antivenoms, which do not efficiently block tissue necrosis and other local effects of B. jararaca or B. jararacussu venoms  at clinical doses [Bothrops in Brazil [L. muta snake by other sulfated polysaccharides obtained from seaweeds was previously reported for sulfated galactans, and was correlated with possible interactions due to the negative and positive charges present in sulfated galactans and in the proteins present in snake venoms, respectively [Fucus vesiculosus exhibited a protective effect against the cytotoxic and myotoxic effects of myotoxins from crotaline snake venoms [The present work demonstrated, for the first time, a sulfated heterorhamnans from a marine alga (Go3) being able to neutralize some of the in vivo activities  induced by al doses . Howevern Brazil ,37. Withectively ,29. Addie venoms ,31, with2+ or Zn2+. These metals should be considered because of their anionic charges, as well as their molecular weights. Most of the active enzymes in venoms require divalent metals to induce their toxic activities [Bothrops or Crotalus, as their venom composition is quite similar. The structural features  are critical for polysaccharides to display pharmacological activities [In particular, the inhibitory mechanism of action of Go3 should be investigated through binding to divalent metals such as Cativities . Howevertivities ,22,38,39tivities . The littivities , and couL. muta and B. jararaca were kindly supplied by Fundação Ezequiel Dias (FUNED), Belo Horizonte, Minas Gerais State, Brazil, and stored at −20 °C until use. Balb/c mice (18–20 g) were obtained from the Núcleo de Animais de Laboratório (NAL) of the Federal Fluminense University (UFF), and were housed under constant temperature (24 ± 1 °C) and light conditions. Experiments were approved by the UFF Institutional Committee for Ethics in Animal Experimentation (protocol number 25), which are in accordance with the guidelines of the Brazilian Committee for Animal Experimentation (COBEA). All reagents were of the best available grade.The venoms of G. oxysperma (Kützing) K. L. Vinogradova ex Scagem et al. were collected on the southern coast of Brazil . An exemplar specimen was deposited at the herbarium of the Federal University of Paraná  under the herbarium identification code UPCB-58059.Specimens of G. oxysperma was extracted twice with water  at 25 °C for 4 h under mechanical stirring. After centrifugation, the supernatant was treated with ethanol (EtOH) , and the EtOH- precipitated material was redissolved in distilled water, dialyzed (cut-off at 12–14 kDa), concentrated by rotary evaporation under reduced pressure, and freeze-dried, giving rise to the polysaccharide extract named “Go1” . This entire process was repeated to create Go2 extract (0.6% yield). The algal residue obtained from extractions at 25 °C was then submitted to similar aqueous extraction at 80 °C, giving rise to the Go3 fraction (13.8% yield). The sulfated heterorhamnans Go3 (10 mg) was resuspended in 1 mL of saline, aliquoted, and stored at −20 °C until assays were performed.The algal material was cleaned, washed with tap water, sun-dried, and milled. The polysaccharides were extracted as previously described ,27. BrieL. muta or B. jararaca venom was determined by indirect hemolytic test using human erythrocytes and hen’s egg yolk emulsion as substrate that contains phospholipids [v/v) was added to the reaction medium and tubes were incubated for 1 h at 37 °C. At this moment, lysolecithin formed during the enzymatic reaction lyzed cells, tubes were centrifuged at 2000 rpm for 10 min, and released hemoglobin was measured at Absorbance 578 nm. After creating a concentration-response curve, the amount of venom (μg mL−1) capable of inducing 100% of hemolysis was denoted as the minimum inhibitory hemolytic concentration (MIHC). Then, one MIHC of L. muta (12 μg mL−1) or B. jararaca (24 μg mL−1) venom was incubated with Go3  for 30 min. at 25 °C, followed by the hemolytic test. Control experiments were performed by incubating venoms with saline in the absence of Go3, or by adding Go3 alone to reaction medium.The degree of hemolysis caused by holipids . After iL. muta or B. jararaca venom was determined as in [w/v in 20 mM Tris-HCl, 8 mM CaCl2, pH 8.8) with modifications. Different venom concentrations were incubated with 0.4 mL azocasein at 37 °C for 90 min in a total volume of 1.2 mL. The enzymatic reaction was stopped by the addition of trichloracetic acid . The tubes were centrifuged at 15,000× g for 3 min. The supernatant was then removed and mixed with 2 M NaOH, and the tubes were read at Absorbance (A) 420 nm to measure the release of azo dye. The effective concentration (EC) was defined as the amount of venom (μg mL−1) able to produce a variation of 0.2 units at A 420 nm. For the inhibitory experiments, one EC of each venom (5 μg mL−1 for L. muta and 10 μg mL−1 for B. jararaca) was incubated with Go3. Proteolytic activity was then determined accordingly.The proteolytic activity of ed as in , using aB. jararaca or L. muta venom were added to plasma, and coagulation time was monitored using a digital Amelung coagulometer . The amount of venom (μg mL−1) able to clot plasma around 60 s was called minimum coagulation dose (MCD), and venom  was incubated with different concentrations of Go3, resulting in venom:Go3 ratios (w/w) of 1:10, 1:20 or 1:50 for 30 min at 25 °C. Following incubation, this mixture was added to the reaction medium and coagulation was monitored as previously described above. Controls were performed by adding Go3 or saline with plasma in the absence of venom.Different concentrations of L. muta (24 µg/mouse) venom was administered subcutaneously (s.c.) into the abdomen cavity of mice; (b) Go3 (140 µg/mouse) or saline was administered intravenously (i.v.); or (c) L. muta venom was administered s.c., and Go3 was injected i.v. 30 min later. Regardless of the experimental group, animals were euthanized after 2 h and blood was collected by cardiac puncture using citrate as anticoagulant. Blood was centrifuged at 1800 g for 10 min, and plasma was transferred to plastic tubes. Then, prothrombin time (PT) or activated partial thromboplastin time (aPTT) tests were performed according to the manufacturer’s instructions . For PT, an aliquot (50 μL) of plasma was incubated for 1 min at 37 °C, and then 100 μL of pre-warmed thromboplastin with calcium were added to initiate coagulation. For the aPTT test, plasma was incubated for 1 min at 37 °C with 100 μL of the aPTT reagent and cephalin plus kaolin for a final volume of 200 μL. Coagulation was then triggered by adding CaCl2  and monitored on the coagulometer.Three different experimental groups were performed, as follows: (a) L. muta or B. jararaca venom were quantified using a procedure described by [B. jararaca or L. muta venom was injected intradermally (i.d.) into the abdominal skin of mice, and animals were euthanized by decapitation 2 h later. The abdominal skin was removed, stretched, and inspected for visual changes in the internal aspect in order to localize hemorrhagic areas. One minimum hemorrhagic dose (MHD) was defined as the amount of venom (µg/mouse) able to produce a hemorrhagic halo of 10 mm, which was 12 and 15 µg/mouse for L. muta and B. jararaca venom, respectively. To analyze the effect of Go3 on venom-induced hemorrhaging, three protocols were employed: (a) incubation: Go3 was mixed with L. muta (24 µg/mouse) or B. jararaca (30 µg/mouse) venom for 30 min at 25 °C, and the mixture was then injected i.d. into mice; (b) prevention: Go3 was administered orally or i.v., and venom was injected s.c. into mice 15 min later; (c) treatment, venom was injected s.c., and Go3 was administered orally or i.v. 15 min later. Additionally, venom was injected s.c., and Go3 was injected s.c. at the site of venom injection 30 or 60 min later.Hemorrhagic lesions produced by ribed by , with soL. muta venom was injected s.c., and Go3 was administered i.v. 1 h later. After 6 h, animals were euthanized and hemorrhagic activity was assessed; (b) L. muta venom was injected s.c., and Go3 was injected i.v. 1 or 2 h later. After 6 h, animals were euthanized and hemorrhagic activity was assessed. Negative controls were performed by injecting solely saline or Go3 instead of venom. The volume of sample injections into mice was 100 µL.Moreover, two additional sets of experiments were performed: (a) L. muta or B. jararaca venom into the right paw, while the left paw received 50 μL of saline. Then, edema was evaluated 1 h after injection and expressed as the percentage increase in the weight of the right paw (ca. 50%) compared to the left one. The effect of Go3 was evaluated by mixing venoms with Go3 for 30 min at 25 °C, followed by the mixture (50 μL) being injected s.c. into animals. Additionally, the protocol for treatment was employed, in which L. muta or B. jararaca venom was injected s.c., and Go3 was injected i.v. 15 min later. Edema was then measured. The control group was injected with Go3 or saline in the absence of venom.The edema-inducing activity of venom was determined according to  with somB. jararaca venom (14 μg/mouse) with Go3 (140 μg/mouse) for 30 min at 25 °C, followed by the mixture being injected i.p. into mice. Positive groups received B. jararaca venom mixed with saline, while the negative control received solely Go3 or saline instead of venom. Similarly, two other protocols were also performed: (a) treatment: B. jararaca venom was injected i.p., and Go3 was injected via the same route 15 or 30 min later; and (b) prevention: Go3 was injected i.p., and B. jararaca venom was injected i.p. 15 or 30 min later. After the end of injections, the number of deaths was observed over a period of 24 h. The injection volume of samples was 100 μL.The antilethality assay was performed by incubating L. muta venom or saline was injected (50 μL) intramuscularly (i.m.) into mice under the right tibial anterior muscles, so that the injected site was positioned just over the extensor digitorum longus (EDL) muscle [L. muta venom (15 μg mL−1) was incubated with Go3 (140 μg mL−1) or saline for 30 min at 25 °C, and the mixture (50 μL) was then injected i.m. into mice; (b) treatment: L. muta venom (50 μL) was injected i.m., and Go3 (50 μL) was injected i.m. at the venom injection site 15 min later. The negative control was performed by injecting saline instead of Go3, and myotoxicity was determined 2 h later, as described here.) muscle . After 2) muscle . Myotoxit-test. p values < 0.05 were considered significant.Results were expressed as the mean ± SE obtained from the number of animals or experiments performed. The statistical significance of differences among experimental groups was evaluated using the Student’s B. jararaca and L. muta venom. These results suggest that the use of Go3 as medicine or phytotherapy to improve antivenom therapy could prevent or diminish the effects of local necrosis that cause severe disabilities, disfigurement, or mortality among snakebite victims. In order to achieve this goal, the development of dosage forms  using Go3 should be investigated further.Overall, Go3 appeared to inhibit the primary toxic activities of


Adenoma miss rate (AMR) has been calculated in several tandem colonoscopy studies, but it costs overmuch to carry out a clinical trial.We aimed to put forward AMR by taking advantage of retrospective data, and to judge the comparability between AMRs from prospective and retrospective data.χ2 test and multivariable logistic regression.Data of the patients accepting repeated colonoscopies during January to September 2016 was retrospectively collected and analyzed. Information was recorded, including bowel preparation quality of the first colonoscopy, size, location, histology and whether missed within the first colonoscopy of each single adenoma. AMR was compared by different risk factors through P < .05). In multivariable logistic regression analysis, adenoma-related factors , and patient-related factors  were found to be independently associated with missing adenomas (P < .05).Around 267 adenomas were detected during 309 pairs of repeated colonoscopies, of which 66 were missed during the first colonoscopies. AMRs of the lesions small in size, nonadvanced in histology, in poor bowel preparation context and located in the proximal colon, were significantly higher than the opposite ones, and old age and male were related to adenoma missing (AMR of retrospective data is comparable to that of tandem studies. Several risk factors influence AMR dramatically, which should be paid attention to. Researchers have found adenoma detection rate (ADR) is closely associated with interval CRC, and ADR is now regarded as the core index of colonoscopy quality.Detection and removal of adenomas by colonoscopy is proven effective in reducing the incidence of colorectal cancer (CRC), and colonoscopy is widely recognized as the gold standard for CRC screening.–7 and a systematic review gave the pooled AMR of 22% in 2006. However, to carry out tandem exam is time and money-consuming, and it is not appropriate to perform tandem exam in clinical practice due to medical ethic issues, so its utility is largely limited. Till now, very limited data of miss rate have been reported and most of them are from North America and European countries. It is not clear how is the situation in China, where morbidity of CRC is increasing dramatically.In order to acquire detail information about missed adenomas and to supplement ADR, adenoma miss rate (AMR) has been brought out. To calculate AMR, tandem colonoscopies, which means 2 colonoscopies with polypectomy taken in the same patient within very short time, must be carried out. According to this examination, AMR refers the proportion of adenomas missed by the first colonoscopy to total adenomas. Former researches reported an AMR of 20% to 47.9%,Here we report AMR in China by analyzing the data of shortly repeated colonoscopies in a tertiary gastrointestinal (GI) endoscopy center. To our knowledge, this is the first report of AMR in the context of routine clinical practice in China, so that we are able to have an idea about how is our colonoscopy quality.22.1This study was undertaken in a tertiary medical center GI endoscopy unit . All patients were referred from outpatient and inpatient of the same medical center. They all completed a standardized tick box questionnaire on main complain and lower GI symptoms, so the indications for colonoscopy were recorded and available for latter research. All the information about patients’ age, gender, indications, images of endoscopic examinations, and endoscopic findings were all recorded in an endoscopic database  prospectively, after obtaining informed consent. Together with pathological information from the department of pathology, the information we needed was ready for referring.This study was approved by the Medical Ethics Committee, Changhai Hospital, Naval Medical University.2.2To collect repeated colonoscopies of same patients within limited time frame to minimize the possibility new adenomas emerging, we search the database for all the colonoscopies during January 2016 to September 2016. Patients were 18 years or older, and during this period they received total colonoscopies (cecum insertion) twice at least, of which the first one was not therapeutic. People who suffered from designated illness , received colectomy or received repeated colonoscopies with interval longer than 6 months were excluded. The inclusion process was shown in Figure 2.3 GI assistants helped with the procedure  when necessary. Polyps removed were sent for pathologic exam.Patients were permitted to eat low-residue foods on the day before colonoscopy. We prescribed 2 L of polyethylene glycol (PEG) 4000 electrolyte powder  or 50 g magnesium sulfate the night before the procedure. All colonoscopies were performed by experienced endoscopists, all of whom had at least 3 years’ experience in colonoscopy. We performed colonoscopy by using Olympus colonoscope . Patients were conscious during colonoscopy or received sedation by using intravenous propofol. Cecal insertion was achieved when ileocecal valve was seen directly. Bowel preparation quality was assessed as excellent, good, fair, poor or inadequate according to Aronchick scale.2.4 Histology details of adenoma includes tubular adenoma, villous adenoma, sessile serrated adenoma, and polyps with low- or high-grade intraepithelial neoplasia. Advanced adenomas are defined as adenomas larger than 1 cm, with villous component, high grade intraepithelial neoplasia or invasive cancer. Locations were grouped as cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum from the proximal to the distal. To explore the influence of location on miss rate, the total large bowel was divided by different points .According to former researches’ classification, lesions were grouped as diminutive (1–5 mm), medium (6–9 mm) and large (10 mm or larger). AMR is compared by different size, histology (advanced or not), bowel preparation quality, and location. Miss rate by patients is also calculated, which is defined as the proportion of subjects who has adenomas not detected in the first colonoscopies. Factors related to patients including age, gender, and bowel preparation quality are also compared. Adenoma detection rate is also calculated.AMR is the main outcome, which refers to the proportion of adenomas missed during first colonoscopy to the total adenomas found during either colonoscopy.χ2 test. Continuous variables are expressed as median and ranges or mean ± standard deviation (SD), and compared with the Student's t-test. Variables that were predictive at the 0.05 level using univariate analysis were entered into the final multivariate analysis, and multivariate logistic regression was performed to identify risk factors for missing adenomas. Two-tailed P-values <.05 were considered statistically significant, and all statistical analysis of the data was performed with Microsoft Excel , and SPSS Statistics 19 .Categorical data are presented as number and percentage and compared by 33.1A total of 2921 repeat colonoscopies were recorded during research time frame. After excluding colonoscopies failed to insert cecum or of which patients had relevant history or other excluding factors, 618 colonoscopies (309 pairs) were retrieved. No severe complications happened. The median age of these patients was 58.5 years (range 20–87), and 62.4% were males. Indications of the first colonoscopy included screening, surveillance and other reasons , and the reasons for a shortly repeated colonoscopy included endoscopic polypectomy after a positive colonoscopy, surveillance after polypectomy, reexamination after a colonoscopy of inadequate bowel preparation, re-examination by a senior clinician after a colonoscopy finding a colonic neoplasm and so on. Bowel preparation quality was grouped as Good (Aronchick's Excellent and Good) and Poor (Aronchick's Fair and Poor). The characteristics of the entire patient population are shown in Table Notably, there were total 35 endoscopists involved in the analysis, and the endoscopists of the 1st and 2nd endoscopies were the same doctors in 71 patients .3.2There were totally 877 polyps detected in either colonoscopies, and 232 polyps were only detected in the second exams, so the overall polyp miss rate (PMR) is 26.5%. Around 267 adenomas were detected during the 618 colonoscopies, of which 201 adenomas in 109 patients were detected in the first colonoscopies and 66 adenomas were missing. So ADR was 35.3% (109/309), and AMR was 24.7% (66/267). Among the 267 adenomas, there were 120 diminutive, 71 medium and 76 large ones. Data of all the adenomas are shown in Table P < .001), nonadvanced (P < .001), poor bowel preparation (P = .028), and different location (P = .016) were associated with missing adenoma. In multivariate analysis, diminutive in size , poor bowel preparation  and located in ascending colon , transverse colon  or sigmoid colon  were also found to be independently associated with missing adenoma. Furthermore, by different dividing points, we found AMR was significantly high in cecum and ascending colon (P = .028) and low in rectum (P = .018).Risk factors related to missing adenoma between the detection and missing groups are summarized in Table P = .006), male sex (P = .014) and poor bowel preparation (P = .006) were associated with missing adenoma. In multivariate analysis, all the 3 variables, ≥60 years , male sex  and poor bowel preparation , were also independently associated with missing adenoma.As to risk factors related to patient, analyses are summarized in Table 4 CRC is one of the very cancers which are suitable for screening, and the colonoscopy-led screening program has been shown to reduce colorectal cancer incidence and mortality in the United States and other developed countries. But organized screening programs are still to be implemented in most countries. In China, CRC screening is still at the early stage, and community-based screening programs are in progress in some places like Shanghai, Tianjin, and Guangzhou. Questionnaire survey and fecal occult blood test are the primary methods rather than colonoscopy due to the short of medical resources and the poor compliance of colonoscopy, and screenees with positive results of primary test will be sent to endoscopy centers for further examinations. Chinese researchers are working on developing a product of stool DNA test of methylated syndecan-2 for colorectal neoplasia screening, and the application prospect is promising.The only sure way to reduce cancer is through screening and healthy lifestyle. While the truth is adenomas are commonly missed during colonoscopy with a poor AMR of 22%. Though highly recommended, colonoscopy is not a perfect method in adenoma detection. Actually, many endoscopists have missed many polyps during colonoscopy, and here we report an overall AMR of 24.7%. So it is an important issue to improve the quality of colonoscopy and not to ignore adenomas during colonoscopy before a better method which can take place of it comes out. Quality control is a big issue in colonoscopy research. ADR as the core index in quality control, which has been proven to be directly related to interval CRC, has already been recommended to be used in assessment of endoscopy centers and endoscopists by several national and societies’ guidelines. For example, ASGE guideline has set an ADR of 25% and 15% for male and female American white screenees older than 60 year old as the minimum standard in 2006, and the benchmarks have been updated to 30% for men and 20% for women in 2015.The utility of colonoscopy and endoscopic polypectomy in CRC prevention is highly recommended, and colonoscopy is regarded as the golden criteria in CRC screening.et al. has proven that ADR is necessary but insufficient for distinguishing high versus low endoscopist performance, and a endoscopist of high ADR can still miss many adenomas in patients who already have adenomas detected. We now know that not only identifying a patient with adenomas is important, but also identifying each single adenoma is equally important, because any missing adenoma is a potential CRC in the future, leave out it is within a patient with or without adenomas detected.However, ADR does work, but is not enough for quality control. Wang  retrieved 76 pairs of colonoscopies performed within 4 months from approximate 15000 examinations to calculate AMR, and found an AMR of 12.0%. They also indicated that tandem colonoscopy or repeated colonoscopy could be used to calculate the true 1-yr recurrence of colorectal adenoma when incorporated with 1-yr re-examination. Kasugai et al retrieved the information of 688 patients who accepted repeated colonoscopies within 1 month, and discovered an AMR of 13.9%. Recently, Shin et al analyzed data of referral patients with advanced adenomas to be resected in the referring hospital, and a per-patient miss rate of adenoma of 47.2% was documented. These work shows AMR from retrospective data is theoretically practicable and comparable to that from tandem exams.AMR pays attention to every single adenoma instead of person suffering from adenomas, so it can be more sensitive than ADR. However, to calculate AMR, tandem colonoscopies must be taken, which cannot be routinely performed in daily practice, and previous AMR data are almost from clinical trials. The limitation of AMR utility makes it uncommon in colonoscopy quality control and its importance and superiority are not well accepted by endoscopists. Nevertheless, the endeavor to calculate AMR using retrospective data has been made. Bensen et al have indicated that adenomas in proximal colon are more often flat or depressed and are therefore harder to be detected than pedunculated ones in distal colon. Moreover, in Pickhardt et al's study of location of missed adenomas, 14 (93.3%) of 15 nonrectal missed adenomas were located on a fold and 10 (71.4%) of these were located on the backside of a fold. When the colonoscope comes into proximal colon, difficulty of turning around and looking back makes the problems above emerge. It is well-reasoned that AMR of proximal adenomas is higher than that of distal ones. Our data also support the viewpoint. Other risk factors such as small in size, nonadvanced in histology, poor bowel preparation, old age and male are also associated with adenoma missing, and the attempts to reduce AMR may follow these cues.In our study, the overall AMR is 24.7% and the AMR of the cecum and ascending colon is 35.6%, and the data indicates proximal adenomas are prone to be missed. Soetikno et alTable The inherent drawbacks of retrospective analysis cannot be avoided. Confined data resource only provide us limited factors for analysis, also with compromised data quality. We are unable to put forward factor analysis other than size, location and bowel preparation quality. We routinely use PEG4000 or magnesium sulfate for bowel preparation, but the retrospective data failed to provide us the difference on AMR between these 2 regimens, because the information was not always recorded completely. However, the difference might exist when these 2 regimens brings different bowel preparation quality. Procedure time has been proven to influence ADR due to doctors’ fatigue, and it may also influence AMR. But the difference exists in afternoon colonoscopies performed in full-day blocks by same endoscopists when ADR is analyzed, we herein cannot do this analysis based on the retrospective data. The same situation happens to the influence of withdrawal time on AMR, which we believe must be solved by randomized controlled trials. The proper of sample size is worth discussing. Longer enrollment timeframe provides larger sample size, but brings the possibility of adenoma emerging rather than missing. Our research is a preliminary one, and further prospective studies concerning the risk factors will surely bring us more valuable information, how to decrease AMR in the proximal colon, for example.We thank the subjects for their participation. We appreciate very much for all the endoscopists and pathologists in Changhai Hospital for their hard work. The last but not the least, we appreciate Dr Kai GU and Dr Yangming GONG (Shanghai CDC), Dr Jiaxin LI , and Dr Yufeng Chen  for sharing the information about CRC screening in their corresponding fields.Conceptualization: Zi-Ye Zhao.Data curation: Cheng-Long Wang, Zhi-Ping Huang, Kai Chen.Formal analysis: Cheng-Long Wang, Zhi-Ping Huang, Kai Chen.Funding acquisition: En-Da Yu.Investigation: Cheng-Long Wang, Zhi-Ping Huang, Kai Chen, Fei-Hu Yan, Liang-Liang Zhu, Yong-Qi Shan, Yong-Jun-Yi Gao, Bai-Rong LI.Methodology: Zi-Ye Zhao.Project administration: En-Da Yu.Resources: Fei-Hu Yan, Liang-Liang Zhu, Yong-Qi Shan, Yong-Jun-Yi Gao, Bai-Rong LI.Software: Zi-Ye Zhao.Supervision: Hao Wang, En-Da Yu.Validation: Hao Wang, Zi-Ye Zhao.Visualization: Zi-Ye Zhao.Writing – original draft: Zi-Ye Zhao.Writing – review & editing: En-Da Yu, Zi-Ye Zhao.Zi-Ye Zhao orcid: 0000-0002-0243-2873