Organophosphate-based pesticides, such as diazinon, are among the most toxic organic contaminants to human and environment. Effective removal of diazinon from contaminated water sources is critical. Zirconium Metal−organic frameworks (Zr-MOFs) are promising candidates for the removal of organic contaminants from wastewater. Herein, we report the adequacy of a bio based Zr-MOF named MIP-202 for the removal of diazinon from water. On the other hand, the use of these materials in powder form is not workable, the development of scalable and economical processes and integrative of these materials onto beads is paramount for industrial processes. Hence, it was reported a scalable, bio aqueous solution-based preparation strategy for Bio Zr-MOF beads production. The composite material exposed identical reactivity under the same ambient parameters compared to powdered material in an aqueous solution. These results signify a critical procedure to an integrated strategy for organophosphates removal using bio-based MOFs, which demonstrates high potential for manufacturing applications such as continued removal of organophosphates from wastewater supplies. Agriculture and industrial manufacturing have contaminated freshwater sources with numerous inorganic and organic pollutants ,2. FindiMany strategies have been explored for diazinon removal from wastewater including photocatalytic degradation and ozonation, coagulation, and chemical and biological oxidation ,14. HoweRecently, significant scientific attention was focused onto the hierarchically ordered mesoporous materials, as they are characterized by their high surface areas, uniform porosity, selectivity, storage capability, mass transport, and diffusion efficient rate . It is eHerein, we chose to investigate the efficiency of an amino acid-based MOF named MIP-202 as a bio-based zirconium MOF for the adsorption of diazinon from water. Notably, the MIP-202 is constructed from Zr-carboxylate bonds which have been highlighted for great interest due to their excellent stability . In addiHowever, powdered forms of MOFs are typically poorly processable for practical applications; thus, formulated forms of MOFs are paramount. Many strategies have been developed to obtain processable forms of MOFs . Howeverl-Aspartic acid , chitosan , sodium alginate , calcium chloride  and diazinon  have been used as it is.Thee starting chemical reactants Zirconium Chloride , Ethanol , 4  and l-aspartic acid  and completely dissolving them in deionized water (10 mL). Then, the solution was transferred into a 100 mL round flask and refluxed at 90 °C under continuous stirring for 24 h. subsequently, the solution was kept out to cool down at room temperature and then washed several times with ethanol. Finally, the resulting precipitate was isolated and centrifuged to obtain the resulting precipitate. The white powder was collected and dried in a vacuum oven at room temperature for 24 h.MIP-202 powder was synthesized using a modified green method to obtain colloidal stable nanoparticles. It is worth mentioning that MIP-202 is prepared without using any organic solvents that commonly used for MOFs synthesis. MIP-202 was synthesized by mixing ZrCl2 in 100 mL of 0.1 mol/L chitosan aqueous solution. Then, the sodium alginate aqueous solution was dropped into the processing solution. After modelling  analysis were done using a D8 Bruker X-ray powder diffractometer,  at 40 kV and 40 mA and intensity data for 2θ from 20° to 70° over a period of 30 min. Fourier-transform infrared spectroscopy (FTIR) spectra were produced using Bruker Vertex 70 to explore chemical properties of MIP-202 after and before polymeric blend immobilization. Scanning electron microscopy (SEM) images were investigated using a Hitachi SU8030 FE-SEM  microscope. The samples images were performed by transmission electron microscope (TEM) JEOL JEM-2100 200 kV  by drop casting the MIP-202 powder and grounded MIP-202/CA composited beads and ethanol onto the 200-mesh copper TEM grid. The apparent surface areas were determined from nitrogen adsorption–desorption isotherms collected at 77 K by a Micromeritics Tristar II 3020. Thermal stability of the samples were performed using thermogravimetric analysis (TGA) as the samples were heated room temperature-700 °C with a rate of 10 °C/min under a constant flow of air. Freeze drying was performed with a critical point freeze dryer. Briefly, freeze drying was used in activation the water-soaked samples over a period of eight hours.The diazinon adsorption efficiency of the MIP-202 bio-MOF/CA beads were investigated using batch technique. In batch method, 100 mg of different materials  was mixed with 100 mL of 50 ppm diazinon solution concentration of pH = 7 at 22 °C for various time intervals (0–90 min) using a shaking incubator . The influence of processing parameters on the adsorption behavior of the MIP-202/CA beads including MIP-202/CA beads dosage (0.1–4.0 g/L), and initial pollutant concentration (10–100 ppm) were constituted. The accuracy of the data was proved by repeating all tests in triplicate and it was used the mean average values in the analysis processes.After the diazinon adsorption processes, the adsorbent materials were separated using the centrifuge to control the final diazinon concentration via high-performance liquid chromatography . Subsequently, the supernatants were filtered using 0.25 µm syringe filters before injection into C18 column. The mobile phase was a mixture of methanol (88%) and ultrapure water (12%). The injection volume of diazinon solution was 20 µL. The flow rate and temperature of operation were 1 mL/min and 30 °C, respectively. Additionally, the detection wavelength of diazinon was 254 nm. Furthermore, the intermediate products of diazinon were set using liquid chromatography tandem mass spectroscopy (LC-MS/MS) analysis. A mixture of 65% methanol: 35% of 0.1% formic acid was utilized to detect the intermediates products of diazinon. The analysis experiments were done at flow rate and temperature of 0.4 mL/min and 35 °C, respectively.oC is the initial diazinon concentration (mg/L); and eC is the diazinon concentration at equilibrium in aqueous solution (mg/L). The adsorption capacity (mg/g) of the diazinon was calculated by Equation (2) [V is the diazinon solution volume (L); and m is the mass of adsorbent material (g).The diazinon removal percentage via the adsorbent materials was determined from Equation (1) .Removaltion (2) :qe = V (eq (mg/g) is the adsorbed amount of diazinon at equilibrium; eC (mg/L) is the concentration of the adsorbate at equilibrium; and mq (mg/g) and LK (L/mg) are Langmuir constants referred to the maximum monolayer adsorption capacity and adsorption energy, respectively. Moreover, the following Freundlich linear equation was applied to analyze the equilibrium data, by plotting log eq versus log eC.FK and nF are Freundlich constants related to the adsorption capacity and intensity, respectively.Langmuir, Freundlich, and Temkin models were used to explain the performance of the adsorption processes of diazinon onto synthesized MIP-202 bio-MOF/CA beads. The Langmuir equation can be presented as following: A (L/g) is the Temkin constant; and B = RT/b (J/mol) is constant referred to adsorption heat.Temkin isotherm model was expressed for the adsorption behavior of diazinon onto the fabricated MIP-202 bio-MOF/CA beads as following .qe = B eq and tq (mg/g) are the amounts of adsorbed diazinon at equilibrium and at time t (min), respectively. k1 (min) expresses the constant rate of the first-order kinetic model. Moreover, the pseudo-second order kinetic model was used to analyze the adsorption kinetic data which can be expressed as following:k2 (g/mg·min) is the constant of the second-order rate. Furthermore, the following equation express Elovich model:α is (mg/g·min) the initial adsorption rate and β (g/mg) refers to the degree of the surface concealment and physical activation energy of adsorption. α and β can be obtained by calculating the slope and interrupt of the linear plot of tq against ln t, respectively. In a similar manner, the intraparticle diffusion affecting the diazinon adsorption processes from aqueous solution was expressed using Weber and Morris equation;ik is the intraparticle diffusion rate constant. The value of C gives prediction about the boundary layer thickness. If intraparticle diffusion occurs, tq vs. t0.5 is linear and if the plot cross through the origin, the rate determination is only due to the intraparticle diffusion.To examine the diazinon adsorption technique on MIP-202 bio-MOF/CA composite beads from wastewater, the pseudo-first order, pseudo-second order, Elovich, and intraparticle diffusion kinetic models were utilized. The Lagergren first-order equation is represented as follows:To evaluate the economic feasibility of the fabricated MIP-202 bio-MOF/CA beads in water treatment processes, the adsorption–desorption cycle was repeated for five times, where the adsorbent material was washed with distilled water and ethanol then dried in air for to be reused.6(μ3-O)4(μ3-OH)4 node and the L-aspartate ligand which explain extraordinary water stabilization due to Zr(IV)−O bonds [−1 in addition to the characteristic peaks of NH2 at 3380 cm−1 and 3495 cm−1 which decorate the internal pores of MIP-202 structure coming from the aspartic acid ligand. Accordingly, both PXRD and FTIR results showed the proper successful incorporation of MIP-202 powder with CA beads.MIP-202 MOF was selected as a bio-based zirconium MOF for the removal of diazinon from wastewater. MIP-202 is constructed from zirconium metal and aspartic acid through strong Zr(IV)−O bonds in which the 12-connected Zr−O bonds . So, MIP−O bonds . As, MIPFor the formation of MIP-202/CA composite beads, to better get a homogeneous solution of MIP-202 powder with the blended chitosan and alginate polymeric matrix, MIP-202 was prepared in nano size forms as confirmed from the SEM and TEM images a,b. SEM In addition, the colloidal stability of the prepared MIP-202 particles was attained as shown from DLS measurements and zeta potential . The sizMIP-202 powder and MIP-202/CA composite beads was conducted using TGA . It is c2/g for MIP-202 powder and 160 m2/g for MIP-202/CA composite beads, respectively, as evident from 3/g and 2.52 nm for the free MIP-202 powder, respectively. These values were estimated to be 0.029 cm3/g for the total pore volume and 12.24 nm for the average pore diameters of MIP-202/CA composite beads.The nitrogen sorption isotherms of the resulting beads exhibited a detention of the porosity of MIP-202 embedded within the CA beads. The calculated BET surface areas was 180 mThe diazinon adsorption feasibility onto the fabricated MIP-202/CA composite beads from waste solutions onto the synthesized materials was performed using batch technique at room temperature.One of the most significant factors in the adsorption processes for MIP-202/CA composite beads is the dosage due to its influences on the adsorbent beads’ capacity. The effect of the prepared MIP-202/CA beads dosage on the diazinon adsorption processes was evaluated after 40 min. The effect of diazinon initial concentrations was examined after 40 min with the initial concentration starting from 5 to 100 ppm in existence of 0.5 g as an optimum material dosage. In a similar manner, LR) in the range 0 to 1 illustrates the favorable adsorption processes of diazinon via Langmuir model [Fn) is 0.357, which is less than the unity, proving that the adsorption process of diazinon onto MIP-202/CA composite beads is unfavorable. On the other hand, the high value of Temkin correlation coefficient demonstrates that the equilibrium data of diazinon adsorption onto the synthesized composite bio-beads are fitted with Temkin isothermal model. Consequently, the Langmuir and Temkin models are the best favorable models for representing the monolayer chemical adsorption of diazinon on the fabricated MIP-202/CA composite beads.The behaviors of adsorption processes of diazinon onto MIP-202/CA beads were investigated by Langmuir, Freundlich, and Temkin equilibrium models as indicated in ir model . HoweverR2 values for the linear plotting of the studied kinetic models for diazinon adsorption onto the synthesized MIP-202/CA beads from waste solutions. It is clear that the linearity of plotting t/tq versus time offers a high R2 for diazinon decontamination from wastewater indicates that the diazinon adsorption processes onto the fabricated MIP-202/CA beads agreed with the pseudo-second order kinetic model. Furthermore, the theoretical and experimental values of diazinon adsorption capacities onto the bio-beads are very close to each other. On the other hand, the correlation coefficient of Elovich equation recorded a high value of 0.921, which proposes that the diazinon adsorption process may be well explained using Elovich model. These data confirm that the studied diazinon removal process onto the fabricated MIP-202/CA beads is mainly controlled by chemisorption process [To examine the diazinon adsorption mechanism onto the synthesized MIP-202/CA composite beads from waste solutions, the presented models of pseudo-first order, pseudo-second order, Elovich, and intraparticle diffusion kinetic models were achieved at  process .The study of regeneration the adsorbent material is one of the most important factors because it effects the total cost of the real applications . Consequmq) of diazinon onto the synthesized MIP-202/CA composite beads were compared with the diazinon adsorption capacities of different adsorbent materials as listed in In this section, the capacity of mono-layer adsorption (In summary, we report for the first time the efficiency of a bio-based zirconium MOF named MIP-202 for the removal of diazinon from polluted water. Since the powder form of MOFs is not applicable in most industrial applications, an eco-friendly and scalable approach for fabrication of bio based composite beads from MIP-202 powder with chitosan and alginate biopolymers named MIP-202/CA beads was developed. The resulting, highly robust, and environmentally friendly MIP-202/CA composite beads demonstrated highly efficient removal of diazinon from polluted water. The maximum adsorption capacity was determined to be 17.77 mg/g and the adsorption behavior of diazinon was fitted with Langmuir model that indicated the monolayer adsorption onto the fabricated MIP-202/CA beads. The eco-friendliness, ease of synthesis, diazinon facile adsorption, and reusability of MIP-202/CA beads make this material a promising candidate to eliminate organophosphate from waste aqueous solutions. Moreover, we believe that our generalizable and scalable approach for integrating bio-MOFs onto bio-based polymers in form of processable bio-beads gives rise to a crucial step for widespread implementation of bio-MOFs material for water treatment industrial applications.


Chronic Obstructive Pulmonary Disease (COPD) is a treatable disease with a high prevalence, and high morbidity associated with significant socioeconomic costs.To carry out a systematic review of the literature to analyze the main cost studies associated with COPD, in order to determine the main factors that influence the costs of the disease.Searches were conducted in PubMed, SCOPUS and Web of Science databases for cost studies on COPD published in English, between the years 2015 and 2020. The search terms were “COPD” OR “pulmonary disease, chronic obstructive”, “cost*” OR “cost of illness”, “economic impact” AND “burden of disease”. The inclusion criteria included the identification of reported cost of the disease, economic burden, medical care expenses or use resources for COPD, the methodology used, data sources, and variables studied.18 publications were analyzed: 17 included direct health costs, 6 included direct non-medical costs, 12 analyzed indirect costs and two reported intangible costs. Most of the studies reported data for developed and European countries, with direct costs being the most studied. Trends were observed in multiple studies of direct and healthcare costs for European countries measured by patient and year, where the higher costs were associated with more severe COPD and a frequent history of exacerbations. The highest costs reported corresponded to hospitalizations and the associated pharmacological treatment. The importance of the loss of productivity and premature retirement within the profile of the COPD patient was also highlighted as the main generator of indirect costs of the disease.COPD generates substantial costs for the health system, mainly related to moderate to severe stages and the exacerbations and complications entailed. It is important to strengthen health systems with monitoring, evaluation and health education models that allow these patients to remain stable to avoid decompensation and subsequent hospitalizations. The main symptoms associated with the disease are dyspnea, chronic cough, and expectoration. These symptoms can often mislead the diagnosis, as they can be confused with other chronic respiratory pathologies with a similar course, such as asthma. The literature generally identifies four phenotypes or clinical forms within the disease that determine different treatments: non-acute, with emphysema or chronic bronchitis, COPD-Asthma, acute with emphysema and acute with chronic bronchitis .The natural history of COPD includes episodes of exacerbation, which become common in the moderate and severe stages of the disease. These episodes increase morbidity-mortality, health system resource utilization (increased costs) and lead to loss of productivity and deterioration of the health-related quality of life (HRQoL) of the patients who suffer from them , 3. ExacEpidemiologically, COPD is identified as the third leading cause of death worldwide, besides representing one of the main causes of increased morbidity, inducing an economic and social burden that is currently on the rise. The morbidity and mortality identified vary from one country to another and from one population group to another, which is why the existing prevalence data widely differ .The main usefulness of cost-of-illness studies is based on the identification and measurement of part of the financial impact of the disease, at the health, social and work levels, helping health system managers with limited budgets and resources to cover a set of unlimited needs and to make a cost-effective allocation between the investment options that exist and are presented to them in their day-to-day activity.In the case of highly prevalent chronic diseases, it is essential to quantify the social and financial magnitude of the disease in all spheres . This will improve the ability to design, prioritize, and apply preventive and health promotion measures, investments in public health or social policies and protocols and care guidelines in the most equitable and efficient way possible. Furthermore, this will also allow constant updating to prevent current practices from becoming obsolete, inefficient, and costly because they are not adapted to the reality of the disease.Various international studies have been carried out to try to estimate the cost of COPD to society or to the health care system, in different manners and applied to different processes of the disease and with highly variable study designs, thus obtaining heterogeneous results. This variability in the methodology used by the different authors makes it difficult to extrapolate results and conclusions, which can lead to a failure to make the most appropriate and cost-effective decisions for managing a disease with an ever-increasing prevalence in society. In view of the wide variety of estimates, it is important to develop a guide that provides a homogeneous estimate and includes comparable data.The principal aim of this systematic review was to identify the main factors that lead to increased costs in COPD patients.A systematic review of the literature on COPD-associated cost studies was conducted and served as the basis for the design of the overall structure of this systematic review. The studies were identified and selected by conducting a literature search using three electronic databases: PubMed, SCOPUS, and Web of Science.The terms included in the thesaurus of the National Library of Medicine of the United States  were used: “COPD or “pulmonary disease, chronic obstructive”, “cost*” or “cost of illness”, “economic impact” and “burden of disease” in the title or abstract fields to retrieve potentially relevant publications in the last five years . The search strategy was limited to publications made in English and Spanish. The results were complemented with a new review with free-text language with keywords of the present monograph and filtering in the last five years in the Google Scholar database after which 36 articles were selected .Publications in English and Spanish.The inclusion criteria consisted of systematic review or original research, meeting the following conditions:Prior to conducting the literature search, a review protocol was developed that included a timeline, the research question, inclusion criteria for the articles to be selected and a table for extracting data from the studies. Figure This review highlights the significant economic burden of COPD. Methodological heterogeneity was found in the studies included in the review. The different authors considered different variables to estimate direct healthcare costs, direct non-medical care costs, indirect costs and intangible costs, and even used different quantification methodologies and different economic conditions, currencies, and billing years. In addition, the variability among the different health systems in which the various studies included in this review are framed must also be considered. In summary, the studies used different approaches to estimate resource consumption. Although each method is valid and appropriate and there is no standard formula to be applied homogeneously in each study, the resources allocated to the disease were identified differently, which can generate disparities in the cost estimation.This disparity leads to a wide variability of results for the cost of COPD disease. Each perspective provides useful and valid information on costs for a particular group (different samples with different patient sociodemographic and disease-related characteristics), although this makes comparison of results difficult.n = 11), systematic review (n = 5) and cross-sectional study (n = 2). These studies were published between 2016 and 2020, thus presenting the most updated data on the subject under study. The monograph is based on a global setting; therefore the analyzed articles were conducted mostly in Europe, but also Asia and America. One of the most important aspects when carrying out a cost study and analyzing its results is to identify the perspective presented by each study. Thus, the present systematic review is made up of studies with a societal perspective (9 articles), followed by the perspective of the National Health System (4 articles), that of the patient (1 article) and that of the public decision-maker (1 article). Finally, regarding the main objective of the research studies analyzed, most studies aim to evaluate the economic and social impact and burden of COPD in the different health systems and societies where the studies are conducted, as well as to identify the main factors that increase the cost of the disease. These are the same objectives sought by the present monograph. See Table First, the most relevant characteristics identified in the eighteen selected articles are presented. The studies selected in this systematic review included the following designs: cohort study , cultural differences and differences in health practices, recall and selection bias (which can lead to an underestimation or overestimation of costs), difficulties in reaching the sample size, heterogeneity in designs between studies, differences in the level of COPD severity of the patients selected, and the non-inclusion of the costs of exacerbations in some of the articles. See Table As shown in Table The main structure presented by most of the studies starts with an analysis of direct health costs, followed by direct costs unrelated to health, and finally by indirect costs. As traditionally occurs in cost-of-illness studies, the least studied costs were the intangible costs, since only 2 of the 18 studies included these in their research. This is possibly due to the lack of information and records and the difficulty of their evaluation, quantification, and subsequent analysis, since it is very difficult to create a monetary value for aspects such as health-related quality of life (HRQoL) or disability.In short, the heterogeneity of the studies makes it difficult to compare results. In this regard, the cost varies greatly depending on the countries studied, whether they have public or private health systems and whether they are conducted in developed or developing countries. In conclusion, a great variability was found in the costs reported in the studies, where the total cost per COPD patient varies strikingly between geographically distant areas.Starting with direct health care costs, the data are analyzed and presented for the different geographical areas studied. Thus, in Europe (the geographic area where most of the studies in this monograph are set), this cost ranges from high-cost areas such as Norway at €10,701 (€/2019) per patient per year and Belgium at €1963 (€/2019) per patient per year. The average cost established among eight European countries  for total direct costs per patient per year is €6182 (€/2019) . SecondlConsidering the indirect costs (the second most studied), starting again with Europe, Rehman et al. , in theiThe following are direct non-health care costs, these types of costs are present in eight of the 18 articles analyzed in this review, many of which have already been included in the section on direct healthcare costs. There are some exceptions that report total direct costs, making no distinction between health and non-medical costs.Finally, only two authors included intangible costs in their articles. In summary they established that patients with mild COPD spend 0.62 years of their life in disability, for those with moderate COPD this can represent as much as 6 years, and in the case of severe stages of the disease, this figure amounts to 9 years y reporting that the quality of life was worse among patients with COPD than in patients without COPD and also that they had a higher risk of depression during the course of the disease than patients who did not suffer from the disease. See Table COPD is one of the main chronic diseases identified in developed countries. The latest epidemiological data indicate a high growth rate, which is projected even more intensely into the future, mainly related to population aging and smoking, which is present in western societies with worrying figures. Its identified morbimortality is one of the factors that disrupts the economic stability of health institutions, which is one of the basic pillars of a nation, and consequently increases the costs attributable to society at large , 2.Regarding the characteristics of the selected articles Table  and theiIn the case of the economic characteristics of the selected articles Table , one of In view of the objectives of this monograph, it is important to mention that the 18 cost studies identified in this systematic review indicate that COPD significantly increases overall economic costs, thus generating significantly higher excess costs for people with COPD, compared to people without the disease. Regarding the factors that increase costs in COPD patients, after reading and analyzing the 18 articles included in the review, the main link that leads to the greatest increase in costs in this pathology is AECOPD (acute exacerbation of COPD), the main complication, which occurs more frequently and is greater with more severe COPD and cases that are worse controlled. Thus, the patients who generate the most costs within the disease are those who are in a moderate-severe stage of the disease and who, in addition, have the worst control of their pathology. Poor control of respiratory disease in general, and specifically in the COPD patient, is based on poor adherence to treatment and a lack of skill or technique in carrying out the prescribed pharmacological treatment. This, together with the presence of other risk factors such as smoking or obesity and factors external to the patient, such as environmental conditions or aging, lead to an increase in the number of exacerbations, which in most cases can lead to visits to the emergency department or even hospitalizations that generate high costs, which, in addition to the patient’s own comorbidities, can be lengthy. These comorbidities are the second most present factor which the studies refer to the most regarding the increase in costs, since patients with a greater number of chronic diseases generate increased costs, due to a greater consumption of drugs, greater hospital and outpatient visits, the performance of greater functional and laboratory tests, and, directly and indirectly affecting parameters that make up work losses, increased disability, loss of HRQOL or the need for the presence of an informal caregiver to support the patient’s basic care.Finally, a breakdown by type of cost shows that all the authors reached the consensus that the main components of direct healthcare costs are hospitalizations (produced in most cases by AECOPD) and medication to treat these complications, whereas in general the other components  are less relevant. Likewise, regarding indirect costs, the studies identified highlight the importance that work losses can have as a consequence of the disease. These are reported by more than half of the authors included in the literature review. This aspect entails not only a loss for the employer or society as a whole, but also a direct impact on the economic stability of the individuals facing the disease and, consequently, also on their families or persons on whom their economic income depends.In conclusion, given the importance of COPD, both for the health system and for society, it would be advisable to reassess the allocation of resources for the disease by taking into account the study of costs.


Telephone therapist delivered CBT (TCBT) and web-based CBT (WCBT) have been shown to be significantly more clinically effective than treatment as usual (TAU) at reducing IBS symptom severity and impact at 12 months in adults with refractory IBS. In this paper we assess the cost-effectiveness of the interventions.Participants were recruited from 74 general practices and three gastroenterology centres in England. Interventions costs were calculated, and other service use and lost employment measured and costed for one-year post randomisation. Quality-adjusted life years (QALYs) were combined with costs to determine cost-effectiveness of TCBT and WCBT compared to TAU.TCBT cost £956 more than TAU  and generated 0.0429 more QALYs. WCBT cost £224 more than TAU  and produced 0.029 more QALYs. Compared to TAU, TCBT had an incremental cost per QALY of £22,284 while the figure for WCBT was £7724. After multiple imputation these ratios increased to £27,436 and £17,388 respectively. Including lost employment and informal care, TCBT had costs that were on average £866 lower than TAU , and WCBT had costs that were £1028 lower than TAU .TCBT and WCBT resulted in more QALYs and higher costs than TAU. Complete case analysis suggests both therapies are cost-effective from a healthcare perspective. Imputation for missing data reduces cost-effectiveness but WCTB remained cost-effective. If the reduced societal costs are included both interventions are likely to be more cost-effective.Trial registration ISRCTN44427879 (registered 18.11.13). Irritable bowel syndrome (IBS) is a common condition with a prevalence of 10–20% . Given tEconomic evaluations of treatments for IBS have been conducted previously. A study from Sweden compared internet therapy with usual care . At 12-mWe recently conducted a three-arm randomised controlled trial n = 558) comparing telephone-delivered cognitive behavioural therapy (TCBT), web-delivered cognitive behavioural therapy (WCBT) and treatment as usual (TAU) , 8. Both comparinThe study recruited from primary care  and secondary care (three gastroenterology centres) settings in the UK between May 2014 and March 2016. The sample consisted of adults with refractory IBS . Full details of the inclusion and exclusion criteria and flow through the trial are provided in the published protocol and clinical results .Participants were randomised with an independent web-based service at the level of the individual with varying block sizes to receive TCBT plus TAU, WCBT plus TAU or TAU alone. TCBT consisted of a patient CBT self-management manual , six 60-Baseline assessments were conducted with follow-ups at three, six, and 12 months following randomisation. The two co-primary outcome measures were the IBS Symptom Severity Score (IBS SSS) and the Work and Social Adjustment Scale (WSAS). The IBS SSS is a 500 point scale with higher scores indicating greater symptomatology , while tCosts were measured from both a health service and a societal perspective (the latter including family care costs and lost employment costs). TCBT and WCBT costs were based on the number of telephone sessions with therapists and the unit cost of therapist time. This was calculated using a therapy cost of £98 per session reported in the Personal Social Services Research Unit’s annual compendium , adjusteCosts to society were estimated by including informal care costs and those resulting from time off work. Informal care costs were calculated by asking participants to say how much time per week family members and friends typically spent providing help in specific areas due to problems associated with IBS. This time was combined with an average hourly wage rate of £15.73 to represent the opportunity cost of informal care . The numComparisons of costs were made at three, six and 12 months as well as the entire follow-up period, with baseline costs controlled for.  The costs of the interventions were only added to the costs for the whole follow-up period. The cost comparisons used regression models with baseline costs entered as an independent variable along with site variables as stratifiers and with bootstrapped percentile 95% confidence intervals generated around the coefficients. Medication costs were calculated but excluded from the analyses due to substantial missing data.Cost-effectiveness analyses were conducted from a healthcare perspective on cases where data on both costs and QALYs were available. Incremental cost-effectiveness ratios were produced, and uncertainty addressed using cost-effectiveness planes (CEPs) and cost-effectiveness acceptability curves (CEACs). The CEPs and CEACs were produced from incremental costs and QALYs generated from bootstrapped resamples of the original data. The range of threshold values used in the CEAC was £0 to £60,000 .We conducted sensitivity analyses by changing the intervention costs upwards and downwards by 25% and 50%. This was done because in other settings therapy might be delivered by professionals at a higher or lower pay grade. Sensitivity analyses were also conducted by using the minimum wage of £7.50 per hour to calculate informal care and lost days from work (assuming 7.5 h per day) and by using the unit cost of a home care worker (£20 per hour) to value informal care. Furthermore, missing cost and EQ-5D-5L data were imputed using multiple imputation methods on 1000 bootstrapped resamples. We assumed that data were missing at random and that potential predictors of costs and EQ-5D-5L scores (at each follow-up point) were corresponding costs and EQ-5D-5L scores from other time points and also the primary clinical measures (IBS-SSS and WSAS). The imputation was conducted on each bootstrapped resample in turn and used chained equations and predictive mean matching based on the five nearest neighbours. The mean of the cost and QALY differences from the 1000 resamples was extracted and reported.Patient and public involvement (PPI) representatives participated in the trial management group and trial steering committee and were included in all phases of trial design, including planning recruitment and recruitment materials.The sample at baseline numbered 558 and is described in detail elsewhere with a consort diagram of patient flow for clinical outcomes [Service use and lost employment data were available for 186 TCBT, 185 WCBT and 187 TAU participants at baseline. At three-month follow-up the figures were 142 (76% of baseline number) for TCBT, 132 (71%) WCBT and 134 (72%) TAU; six-month follow-up 135 (73%) TCBT, 115 (62%) WCBT and 128 (68%) TAU; 12-month follow-up 130 (70%) TCBT, 120 (65%) WCBT and 130 (70%) TAU. During the six-month period prior to baseline assessment, over four-fifths of participants had contacts with a GP  and WCBT had costs that were £151 higher than for TAU . Mean societal costs  were £682 for TCBT, £723 for WCBT, and £836 for TAU. Adjusting for baseline, TCBT had mean total costs that were £25 higher than for TAU  and WCBT had mean total costs that were £218 higher than for TAU .In the three months up to the six-month follow-up, costs were highest for GPs and other doctors. Average non-intervention healthcare costs were £281 for TCBT, £224 for WCBT, and £206 for TAU. After adjusting for baseline costs TCBT had £86 higher costs than TAU  and WCBT had costs that were £39 higher than for TAU  and WCBT having costs that were £420 lower than for TAU .In the six months up to the 12-month follow-up, average healthcare costs (excluding the intervention) were £519 for TCBT, £325 for WCBT, and £393 for TAU. After adjusting for baseline, TCBT had non-intervention healthcare costs that were on average £144 more than for TAU  and WCBT had costs that were £42 lower than for TAU . The higher level of informal care costs for TAU continued during this period. The TAU group also had higher lost employment costs than the other groups. Mean societal costs  were £1055 for TCBT, £1029 for WCBT, and £2103 for TAU. After adjusting for baseline, TCBT had costs that were £873 lower than for TAU  and WCBT had costs that were £751 lower than for TAU .Mean (SD) total healthcare costs, with ACTIB CBT costs included, over the whole one-year follow-up period were £1650 (£1931) for TCBT, £943 (£955) for WCBT, and £715 (£884) for TAU. Adjusting for baseline, TCBT had costs that were on average £943 higher than for TAU which was statistically significant  and WCBT had costs that were £278 higher than for TAU which was also significant . Cost differences for participants where EQ-5D-5L data were available  were £956  for TCBT and £224 for WCBT .The mean (SD) societal costs over the one-year follow-up period, again including ACTIB CBT costs, were £3065 (£5179) for TCBT, £2094 (£3069) for WCBT, and £4374  for TAU. After adjusting for baseline, TCBT had costs that were £866 lower than TAU  and WCBT had costs that were £1028 lower than for TAU .Response to the EQ-5D-5L was relatively low  TCBT participants, 92 (50%) WBT participants, and 100 (53%) TAU participants. Dividing incremental healthcare costs by incremental QALYs differences for those with both cost and QALY data results in the following incremental cost-effectiveness ratios (ICERs): TCBT v TAU £22,284 and WCBT v TAU £7724. The ICER for TCBT compared to WCBT was £52,662.Uncertainty around the ICERs is illustrated in Figs. After multiple imputation for missing EQ-5D-5L-based utility scores and costs it was found that TCBT had an ICER of £27,436 per QALY compared to TAU. WCBT resulted in an ICER of £17,388 per QALY. When therapy costs were reduced by 25% and 50% the ICERs from the healthcare perspective compared to TAU were reduced for both groups and the ICER for TCBT was now well below the £20,000 threshold  and WCBT had costs that were £389 lower than for TAU . When the cost of a homecare worker was used to value informal care, the mean societal costs were £3281 for TCBT, £2275 for WCBT and £5098 for TAU. After adjusting for baseline, TCBT had costs that were £1228 lower than for TAU  and WCBT had costs that were £1241 lower than for TAU .These cost-effectiveness analyses found that TCBT and WCBT resulted in increased healthcare costs over the follow-up period. Higher costs can be justified if outcomes are improved sufficiently. The complete case analysis revealed that the cost per QALY for WCBT was lower than the threshold often assumed to guide NICE decisions in England . TCBT on the other hand had a cost-effectiveness ratio slightly above this threshold. On this basis, WCBT would be the preferred option.However, there was much missing EQ-5D-5L data. This was due to the requirement placed on the trial to use the EuroQol online website to collect the EQ-5D data which required electronic transfer of participants from the trial data collection software platform to the EQ-5D website to input the data. This did not always work effectively, and so data collection was not always completed. With imputation for the missing data, the ICERs both increased substantially. TCBT would be now more unlikely to be considered cost-effective at such a ratio  but WCBT would still be below the NICE threshold.A strength of the study was the comprehensive approach to costing adopted. Both TCBT and WCBT resulted in lower societal costs than TAU. This was because of reduced informal care from family and friends compared to TAU. There was though no evidence that lost work time differed between the groups. From this societal perspective we may deduce that TCBT and WCBT both dominate TAU (i.e. they are more effective and less expensive). However, the confidence intervals are wide and do not exclude zero cost differences. NICE do not usually consider carer costs in their decision-making process though it could be argued with increasing integration of health and social care services and budgets that these are becoming more important to consider. This is a key strength of the study as interventions such as these seem to have benefits outside of the healthcare system. From an economic perspective all relevant effects should be included and costs computed.Elsewhere, Andersson et al found that internet-based therapy resulted in reduced societal costs for patients compared to participating in a discussion group. . This coThere were limitations to this economic evaluation. First, and most importantly, the quantity of missing EQ-5D-5L data was a concern. Those with missing data had worse IBS-SSS and WASAS scores at each time point and so imputation from these resulted in smaller QALY gains for TCBT and WCBT relative to TAU (details available from authors).Second, service use data were provided by participants themselves and there may have been recall accuracy problems. However, this would not likely affect one group more than another and it was the only option for collecting comprehensive data. Third, medication data were not of high quality. We did know what medications were taken but quantities and durations were not complete. Many medications were bought ‘over the counter’ and exact details may not have always been available. However, these costs would be a small proportion of the total healthcare cost and their inclusion would have only a marginal effect. We might though have expected TAU to result in higher medication costs in the absence of an active therapy.Third, the economic analyses were trial based and so limited to the 12-month follow-up period. It could well be that economic impacts continue beyond this time and modelling to extrapolate these results is warranted.Finally, the intervention costs for WCBT were apportioned over the trial population. If both forms of ACTIB CBT were rolled out over a larger patient population then the fixed costs of the intervention would fall per patient.We believe this to be the largest trial of CBT for IBS worldwide to date and the first to evaluate telephone and web-based CBT in the same trial. It was a rigorously conducted trial with broad inclusion criteria and patients recruited from both primary and secondary care. The trial interventions were provided by NHS therapists in a real-world NHS setting. The inclusion of the cost-effectiveness component provides further important evidence for policy makers.In conclusion, the complete case analysis suggests that both the therapies are cost-effective from a healthcare perspective. This is reduced when using imputation methods, but the web-based CBT intervention remained cost-effective. Both interventions produce important savings in terms of carer time and if this is valued then the interventions do cover their cost. These results add important research evidence to support recommendations in NICE Guidelines that CBT should be offered to people with ongoing IBS symptoms.


Trees and herbs that produce fruits represent the most valuable agricultural food commodities in the world. However, the yield of these crops is not fully achieved due to biotic factors such as bacteria, fungi, and viruses. Viruses are capable of causing alterations in plant growth and development, thereby impacting the yield of their hosts significantly. In this work, we first compiled the world′s most comprehensive list of known edible fruits that fits our definition. Then, plant viruses infecting those trees and herbs that produce fruits with commercial importance in the global market were identified. The identified plant viruses belong to 30 families, most of them containing single-stranded RNA genomes. Importantly, we show the overall picture of the host range for some virus families following an evolutionary approach. Further, the current knowledge about plant-virus interactions, focusing on the main disorders they cause, as well as yield losses, is summarized. Additionally, since accurate diagnosis methods are of pivotal importance for viral diseases control, the current and emerging technologies for the detection of these plant pathogens are described. Finally, the most promising strategies employed to control viral diseases in the field are presented, focusing on solutions that are long-lasting. Since the appearance of agriculture, far earlier than previously thought, a few places around the world were sites of origin for the modern crops we see today ,2,3. In Although everyone is familiar with the traits of fruits, such as increased size of the pericarp tissue and high sugar concentrations, these traits evolved initially for dispersing seeds by means of megafaunal species ,12,13,14Putting aside the shortage of fruits and, consequently, higher prices caused by the COVID-19 pandemic, the biggest challenge faced by fruit crops has always been the range of pathogens that can cause disease ,23,24. AAlthough technically nuts and grains are also fruits, we have restricted our review to trees or herbs that produce fleshy seed-associated structures that typically are sweet or sour and edible when raw . Taking EEFs classified as eudicots corresponded to 438 genera, 119 families, and 38 orders, whereas monocots and magnoliids were classified into 32, 16, and 7 and 15, 5, and 2, respectively . Until rRubus (161), Prunus (112) and Crataegus (74), constituted the largest family of Rosaceae (575), order Rosales (685) (Ericaceae (135) and Solanaceae (92) were also numerous, in which the genera Vaccinium (56) and Solanum (36) were highly represented, respectively (Rosaceae are well known (cultivated), including apples , pears (Pyrus sp.), plums, and cherries (Prunus sp.), among others. However, many more EFFs within the family Rosaceae are found in the wild such as Ghingaroo (Pyracantha crenulata), Mehul (Pyrus pashia), and Rubus niveus. In the case of EFFs classified within the clades of monocots and magnoliids (Musa acuminata and Annona muricata), whereas others remain in the wild  (es (685) . EFFs beectively . In factgnoliids , the samctively) .Prunus, Citrus, Capsicum, Ribes, and Solanum, mainly (Solanum lycopersicum), banana (M. acuminata), apple , and grapes (Vitis vinifera) standing out as the most traded EFFs , which are transmitted by a living organism called a vector, are the major infectious agents that cause plant disease ,57,58. OAvr) genes that interfere or suppress PTI . C. CRutaceiraecola ,159,234.iraecola . In combee years ,159,235. hybrids . Despite hybrids . For exa hybrids ,238. In f 40–98% .Viruses can cause huge economic losses by affecting the quality and productivity of various fruit plant crops like banana, apple, grapevine, citrus, and others . Once inThe first way to detect viral infections was according to the symptoms they produced. Such an approach was based on biological indexing tests, which were time-consuming and unreliable, especially in cases in which the virus infection was latent or plants exhibited virus-like symptoms unrelated to virus etiology ,240. TheExtensive reviews and chapters covering diverse diagnostic methods of viruses infecting plants have been described in the literature ,246,247;Potyvirus, Potyviridae) or CMV, PRSV, and groundnut bud necrosis virus  simultaneously in Cucurbitaceae, Solanaceae, and other hosts [ELISA is a cost-effective and robust assay for the routine detection of plant viruses, especially those present in crude extracts, due to its simplicity, detection limit 1–10 ng/mL, ability to analyze a large number of samples, easy interpretation, and semi-quantitative results ,249. Forer hosts .Sadwavirus, Secoviridae), and plum pox virus  infecting apricot, plum, and peach [Orthotospovirus, Tospoviridae) infecting pepper and tomato plants [Comovirus, Comoviridae), tobacco etch virus , TMV, tobacco ringspot virus , tobacco streak virus , ToLCNDV, ToRSV, ZYMV, CGMMV, melon necrotic spot virus , melon severe mosaic virus , PepMV, pepper mild mottle virus , potato virus X , and PVY, among others [LFA is an ELISA variant that simplified virus identification . LFA cannd peach ,264,265.o plants ,267, ando plants . Finallyg others .Serological assays such as dot immunobinding assay (DIBA) and tissue blot immunoassay (TBIA) also allow simultaneous screening of large numbers of samples . In thesBadnavirus, Caulimoviridae) were identified in silico by comparison with caulimoviruses, retroviruses, and rice tungro bacilliform virus. A purified fusion protein p37 was injected in rabbits as an antigen for raising polyclonal antiserum. The antiserum was successfully used in antigen-coated plate-ELISA for specific detection of BSMYV in banana fields and tissue cultures raised. On the other hand, the antiserum was also utilized in immuno-capture PCR (IC-PCR) for indexing of episomal BSMYV infection [New approaches are emerging in order to improve serological diagnosis. Some involve the identification of epitopes in the viral CP by computer simulation or functionally by epitope mapping and its subsequent artificial synthesis . Accordinfection . This biMolecular diagnostics began in the mid-1980s with the introduction of PCR, and the first PCR method for virus detection was published ten years later . PCR-basNested PCR is useful when the virus titer is very low, the target gene is unstable, and it cannot be checked by electrophoresis due to low amplification product. The product from primary PCR amplification is used for second PCR amplification. Several viruses, including prunus necrotic ringspot virus , prune dwarf virus , PPV, and CTV, were detected by this technique. Co-operational PCR also requires four primers; however, one is external, and three are internal primers instead of two external and two internal primers associated with nested PCR. This technique shows similar sensitivity to nested PCR, detection in real-time, capability of coupling with dot blot hybridization, it can avoid false-positive results shown at nested PCR, and it also can be applied to capillary air thermal cyclers. This technique has been useful to detect cherry leaf roll virus  with higher sensitivity than RT-PCR. Digital PCR does not require any reference standards for nucleic acid quantification but rather produces an accurate quantification. The PCR sample is divided into thousands of nanodrops, and after amplification, the drops containing the sequence of the target DNA are detected by fluorescence as positive and those without fluorescence as negative. Then, the statistical analysis of the number of nanodrops gives an exact count of the target DNA, and this helps count the viral charge. Reverse transcriptase droplet digital PCR detected PMMoV in the presence of qPCR inhibitors [Multiplex PCR, Real-time PCR, and Immunocapture-PCR are described in the following sections. Importantly, PCR has been used successfully for the detection of several viruses that infect citrus in India, including CTV, citrus yellow mosaic virus , and Indian citrus ringspot virus  [Many commercial nucleic acid extraction kits  have replaced complicated and time-consuming conventional nucleic acid extraction protocols ,279,280.hibitors . Finallyviridae) . PCR-basviridae) ,285 as wviridae) .Reverse transcription-multiplex PCR can simultaneously detect various target RNA. After reverse transcription of target RNA, cDNAs are simultaneously amplified with a set of specific primers in a single tube by multiplex PCR. In order to quantify viral load, this technique can be performed in real-time mode, known as reverse transcription-real-time multiplex PCR. Real-time multiplex PCR uses a set of species-specific primers and probe that is labeled with different fluorescent dyes for each target species so that approximately 2–5 species  can be detected simultaneously in a single real-time PCR reaction. Compared to real-time single PCR, real-time multiplex PCR shortens the processing time and reduces the use of reagents. Multiplex RT-PCR has been successfully applied for aphid-borne viruses infecting strawberry, such as strawberry crinkle virus , strawberry mild yellow edge virus , strawberry mottle virus , and strawberry vein banding virus  [Pospiviroidae and the ASGV infecting citrus plants [Multiplex PCR is a technique for simultaneous and sensitive detection of different DNA fragments in one single PCR reaction , and it viridae) . Furthers plants . Anothers plants ,296,297,s plants . One of Reverse transcription-real-time multiplex PCR allows detection of various target RNA through simultaneous amplification of cDNAs, produced by reverse transcription, with a set of specific primers in a single tube by multiplex PCR, producing quantitative results [nested real-time reverse transcription PCR is a simple and sensitive method for the detection of pathogenic microorganisms. This technique involves a previous reverse transcription step to synthesize cDNA and the use of two external and two internal primers that are complementary to the target sequence, which increases sensitivity, providing quantitative results [Mandarivirus, Alphaflexiviridae) [Grablovirus, Geminiviridae) in grapevines [One of the limitations of PCR and RT-PCR for virus detection is that PCR products require agarose gel staining with fluorescent dyes such as ethidium bromide, SYBR Green, SYBR Gold, SYBR Safe, Eva Green, GelRed, EZ-Vision, among others , which i results . On the  results . In summviridae) ,302,303.apevines .BSV worldwide [In this technique, viral particles are captured with virus-specific antibodies, followed by the release of viral nucleic acid for PCR amplification. This is very convenient for the detection of plant viruses in which inhibitory plant compounds or low viral titer hinder PCR amplification ,306. It orldwide ,308. It orldwide , and esporldwide .Bacillus subtilis-derived (Bst) NA polymerase, produce amplicons containing loop regions to which further primers bind, allowing amplification to proceed without thermal cycling [Real-time LAMP is a constant temperature amplification method carried out at 60–65 °C, for which only a simple water bath is required. This technique eliminates reverse transcription steps, as well as PCR instrument cooling time, which shortens the amplification time. Adding a fluorescent DNA intercalating dye into the reaction enables monitoring of a fluorescence amplification curve. Compared to conventional LAMP assays, this method avoids visible error, enables quantitative detection, and is more suitable for multi-sample analysis. Reverse transcription LAMP can synthesize cDNA from template RNA and apply LAMP technology to amplify and detect them. As the template is an RNA sample, in addition to the reagents of DNA amplification, reverse transcriptase is added to the reaction mixture. After mixing and incubating at a constant temperature between 60–65 °C, amplification and detection can be carried out in a single step. Multiplex LAMP consists in simultaneous detection of multiple target genes in LAMP reaction, which increases diagnosis specificity. In addition, reverse transcription LAMP in a single tube can be coupled to multiplex LAMP for detection. Electric LAMP is based on an electronic simulation that provides fast and inexpensive putative tests of LAMP primers on target sequences compatibility. This aids to determine the opportunity of existing primers to detect recently discovered sequence variants. Finally, in-disc LAMP is an integrated device composed of micro-reactors embedded onto compact discs for real-time targeted DNA determination. This method requires similar reagents used in conventional LAMP, and it is performed in a micro-reactor placed in a 65 °C oven. During incubation, the disc is cyclically scanned and optically read to obtain quantitative results. LAMP has been developed and standardized for some of the viruses infecting bananas , citrus (CYMV), grapevines (GLRaV-3), and apples (ASGV) [The PCR has been the main tool for viral detection; however, because of temperature cycling, it is time-consuming and less useful than isothermal methods. The LAMP technique has been developed for specific, sensitive, and rapid nucleic acid amplification. This assay uses a group of 4–6 special primers, which together with the strand-displacement activity of  cycling . The who cycling . LAMP pr cycling . There a cycling . Real-tis (ASGV) ,320,321.s (ASGV) . This prAmpelovirus, Closteroviridae), PPV, tomato mottle virus , and TYLCV, infecting fruit plant crops [In this technique, the isothermal amplification of specific DNA targets is achieved by the combination of proteins and enzymes, such as recombinase, single-stranded binding proteins, and the strand-displacing activity of the polymerase. In combination, all these proteins produce amplicons in 10–15 min at a constant low temperature. There is no need for initial denaturation of the dsDNA target. RPA amplicons can be visualized on gel or by fluorescence and/or hybridization ,324. Comnt crops ,324,326.Badnavirus, Caulimoviridae) [Badnavirus, Caulimoviridae) variants causing the leaf streak disease of bananas in India [Badnaviruses, BSV and sugarcane bacilliform virus , and the Caulimovirus, cauliflower mosaic virus  [RCA is another isothermal amplification method for viral detection in fruit plants. It uses exo-resistant random hexamer primers and the strand displacement activity of Phi29 DNA polymerase  to ampliviridae) . Furtherin India . RCA, hoin India . RCA hadin India ,334. Howviridae) .Closteroviridae members, including GLRaV-4, GLRaV-7, and GLRaV-9, were detected for the first time in Chilean grapevines using an oligonucleotide microarray [V. vinifera, V. labrusca, and interspecific hybrids. The majority of infected vines was singly infected, while some were mixed-infected with viruses from two or more families. Representatives of the four main virus families including Betaflexiviridae, Closteroviridae, Secoviridae, and Tymoviridae were found alone and in combination [Some fruit plant crops are infected by a large number of viruses, as is the case of grapevine infected with more than 80 viruses . In suchcroarray . A largecroarray . Besidescroarray . This arbination . The maibination .NGS technologies, developed in 2005, are massively parallel sequencing platforms that have allowed the rapid identification of viruses and viroids. Today, Illumina technology is the most widely used for sequencing. The process begins with DNA fragmentation and incorporation of adapters that contain segments acting as reference points during amplification, sequencing, and analysis . With thBiosensors are portable diagnostic devices based on antigen–antibody interactions (immunosensors) or nucleic acid hybridization coupled to a physicochemical transducer microsystem . A bioseOrthotospovirus, Tospoviridae) diagnosis, which showed ~1000 times more sensitivity than DAC-ELISA [Machlomovirus, Tombusviridae), and they could be useful to detect viruses infecting fruit plants [The biosensor based on the bioelectric recognition assay (BERA) is an intelligent system for the detection of plant viruses, combining the principle of artificial neural networks and biosensors. For example, BERA biosensors detect the electric response produced by the interaction of a cell culture suspended in a gel matrix with the CGMMV . This reAC-ELISA . The antAC-ELISA . Piezoelt plants ,367.Alphaflexiviridae) and odontoglossum ringspot virus  infections in orchids are more sensitive than those based on antibody recognition [Potyvirus, Potyviridae) infections [An optical miniaturized paper-based DNA sensor identified the early infection caused by BBTV. Some DNA biosensors are based on nucleic acid hybridization and QCM detection of DNA molecules ,369. In ognition . Compareognition . A potenognition . Additiofections .Ophiovirus, Aspiviridae), and mulberry ringspot virus  [Pathogen-free planting material and seeds are fundamental for the fruit industry. Disease-free nurseries should be raised by taking propagative materials from disease-free healthy plants that are identified by making tests of various fruit orchards ,374,375.viridae) .Ampelovirus, Closteroviridae) [Nutrition is an environmental factor affecting plant survival and resistance to diseases, pathogen virulence, and presence of biocontrol agents. Proper nutrition is a preventive measure for fruit plant disease, whereas wrong or excessive nutrient applications can bring problems ,385,386.viridae) . Certainviridae) . In a grviridae) . Tomato viridae) ; howeverviridae) .PRV was significantly lower in dicultures of zucchini and buckwheat, white clover, okra, or sunn hemp than zucchini monoculture during three years [Potexvirus, Alphaflexiviridae) [Intercropping of certain crops helps in reducing the inoculum of the diseases or population of root-knot nematodes in the fruit crops. These crops produce chemical compounds that inhibit the microorganism population, protecting the main crop from disease. The antagonistic plants release toxic substances in soil that help in reducing the population of plant-parasitic nematodes . The peree years . In contee years . Additioviridae) ,397.Nucellar embryony is a form of seed reproduction that occurs in certain plant species, such as citrus trees. Nucellar embryos are produced asexually from somatic cells of seed parents . Since cOrchard roguing is another important strategy for controlling viral diseases in fruit crops. This method is a cost-effective and environmentally safe disease management practice to identify and eradicate infected fruit plant sources in order to avoid the spread of the disease ,407. OncEvodia hupehensis are symptomless carriers of CTV, and they should be eradicated from the orchards as they act as foci for the secondary spread of the virus [Cheravirus, family Secoviridae), such as balsamroot, dandelion, and plantain, reduces the virus incidence [Nepovirus, Secoviridae) are seed-borne of Chenopodium quinoa, and their removal helps in controlling the disease [B. tabaci. In addition, the cultivation of papaya near cucurbits should be avoided in order to reduce the incidence of PRSV [Wild plants and weeds may also serve as reservoirs of both virus and vector . Severalhe virus ,418,419.ncidence . For exa disease . Aside f of PRSV . TherefoPentalonia nigronervosa with insecticidal sprays, dust, injection, or encapsulation in situ [Trichovirus, Betaflexiviridae) and peach mosaic virus  are transmitted by bud mite Eriophyes inequalis, and control of the mite reduces the disease incidence [Begomovirus, Geminiviridae) due to protection from whitefly transmission. On the other hand, the spray of fosmite for whitefly control was effective in reducing the disease incidence [Closteroviridae) [Insect vectors are the main pathway for the dispersal of plant viruses. The injuries caused by insects at the time of virus transmission may also serve as entry points for the penetration of other pathogens. In this regard, the successful control of these diseases relies on the proper management of insect vectors. The BBTV could be reduced by eliminating the aphid  in situ . Additio in situ . Similar in situ ,425. Chencidence . In the ncidence ,428. On ncidence . In addincidence . Sprayinviridae) . Finallyviridae) .Ilarvirus, Bromoviridae) [Heat treatments such as sterilization and solarization have been used for reducing or eliminating the inoculum in propagative plant parts, their products and in the soil ,433. Soiviridae) . Heat thviridae) . Banana viridae) . Additioviridae) . Water bviridae) . Finallyviridae) .Chrysoperla rufilabris in caged watermelon contributed to a decrease in the populations of Bemisia tabaci, the vector silverleaf whitefly of geminiviruses, in Texas [Trialeurodes vaporariorum is the vector of criniviruses (genus Crinivirus) such as strawberry pallidosis associated virus , tomato chlorosis virus , and tomato infectious chlorosis virus , among others [Amblydromalus limonicus and Amblyseius swirskii significantly reduced greenhouse whiteflies densities on greenhouse-grown strawberries [Biological control involves the usage of microorganisms for the control of harmful pathogens causing plant diseases without disturbing the ecological balance ,443. Usuin Texas . The greg others . A biocowberries . On the wberries . Cross pwberries . In Indiwberries ,452,453.wberries . Likely,wberries .PRSV, comparable to Q-2000VI, with satisfactory protection until harvest [Potyvirus, Potyviridae), and TMV, infecting tomatoes [Tobamovirus, Virgaviridae) [The application of chemicals is still the principal method for controlling various plant diseases. An ideal pesticide should be foolproof, highly toxic to pathogens at lower concentrations but safer for human beings or animals. In addition, these chemicals should be cheap, easily available, persistent, easy to spread, easy to handle and apply, stable, and simple to prepare . The che harvest . On the  harvest . In addi harvest : 10% tri harvest . These ttomatoes . Thuja iviridae) . On the viridae) .loci [loci included a candidate gene with similarities to the A. thaliana RTM-2 gene, which is implicated in the restriction of the systemic movement of other potyviruses [Lycopersicum and Capsicum spp., resistance to ZYMV, MNSV, and cucumber vein yellowing virus  in Cucumis melo, and resistance to ZYMV and PRSV in Cucumis sativus. However, redundancy among eIF4E genes can restrict the efficient use of knockout alleles in breeding. Similar strategies can be extended to other plant factors required by viruses, such as small GTP binding proteins (AtARL8a/b/c), required for ToMV replication, DNA binding protein phosphatase (AtDBP1), and DEAD-box RNA helicase-like protein (AtRH8/PpDDXL), required for PPV and turnip mosaic virus  replication in A. thaliana and peach [The use of resistant varieties is the most effective and safest means of controlling crop diseases. Several diseases resistant varieties have been developed in fruit crops; however, there is a lot to be achieved against many important diseases ,464. Anyloci . One of yviruses . Howeveryviruses  and the yviruses . Additioyviruses ,475, CP yviruses , CP fromyviruses . eIF4E tyviruses . Conseqund peach .Finally, the generation of virus-resistant varieties also needs to consider the specific climatic conditions for each growing area in order to increase the yield of crops ,481.Quarantine is the legal restriction on the introduction, movement, and spread of a new pathogen to the disease-free area. With the purpose of preventing the diseases spread to the new unaffected areas, mandatory measures such as bud wood certification, crop inspection, and establishing orchards in regions unfavorable for pathogen are taken ,483,484.Bromoviridae, Betaflexiviridae, Secoviridae and Closteroviridae families. This wide host range distribution, together with the potential co-existence of several viruses in the same host, could lead to the emergence of novel viruses that can threaten the fruit industry in the future. Further, we also have presented the most important approaches for the diagnosis of plant viruses as they are of pivotal importance for the timely use of preventive and protective measures to confine the virus(es) and prevent yield losses. This is particularly important for farmers in developing countries, in which, besides facing policy-related and structural barriers, losses caused by plant viruses are of considerable concern.EFFs are among the most valuable agricultural food commodities, and the world seems unable to get enough of them, as is reflected by their fastest growth rates of exportation in recent years. However, fruit trees and herbs pass through a series of growth steps or stages before fruits fully ripen, which not only demand continuous nutrient supply across all their growth but also make them susceptible to be infected by plant viruses during this time. To our knowledge, here we have compiled the world′s most comprehensive list of known edible fruits that fit our definition. Moreover, EFFs have been classified taxonomically according to the major clades within the plant kingdom. Additionally, plant viruses infecting the tiny number of EFFs with significant commercial importance in the global market were addressed according to an evolutionary perspective of plant evolution, finding that EFFs belonging to eudicots were hosts for the major number of plant virus families. In addition, the genome composition of virus species infecting commercially important EFFs was mostly represented by ssRNA molecules, whereas other forms of nucleic acid genomes were represented in minor proportions. Remarkably, the host range for some plant virus families showed a wide distribution such as Currently, food security is of great concern because the human population is growing, soil fertility is declining, and global warming is changing the weather patterns. Thus, agriculture in general, including EFFs, has to face challenging times in the near future. For instance, COVID-19 outbreaks among workers have challenged the fruit industry by idling processing plants, disrupting supply chains, and farmers have to cope with unprecedented threats to fulfill the demand of EFFs. However, in the age of artificial intelligence, robots and computer vision, the aforementioned issues could be solved by automation. This is what AgTech companies are trying to address with innovative ideas. Therefore, by combining science and technology with agriculture, indoor farming is gaining adherents around the world. Smart farming is not free of challenges, but it could be the solution for humanity because conventional agriculture contributes significantly to the greenhouse gas emissions that are causing climate change. In the case of plant viruses, indoor farming could avoid the impact of these plant pathogens significantly, increasing the yield of crops for an ever-increasing human population. Finally, current fruit tree domestication attempts must consider not only early fructification and high rates of productivity but also the molecular determinants that could confer resistance to viruses.


In 2018, there was an outbreak of African swine fever (ASF) in China, which spread to other provinces in the following 3 years and severely damaged China's pig industry. ASF is caused by the African swine fever virus (ASFV). Given that the genome of the African swine fever virus is very complex and whole genome information is currently inadequate, it is important to efficiently obtain virus genome sequences for genomic and epidemiological studies. The prevalent ASFV strains have low genetic variability; therefore, whole genome sequencing analysis provides a basis for the study of ASFV. We provide a method for the efficient sequencing of whole genomes, which requires only a small number of tissues. The database construction method was selected according to the genomic types of ASFV, and the whole ASFV genome was obtained through data filtering, host sequence removal, virus classification, data assembly, virus sequence identification, statistical analysis, gene prediction, and functional analysis. Our proposed method will facilitate ASFV genome sequencing and novel virus discovery. Asfarviridae and the only known DNA arbovirus , which is the only member of the family rbovirus , 2. Deperbovirus . The virthodoros . ASF wasthodoros , 5, 6. Tthodoros –9. In Authodoros .ASFV has a high genetic and antigenic diversity. Based on the p72 protein (B646L), 24 genotypes have been identified, while at least eight serotypes are recognized based on hemadsorption inhibition , 11. TheWe obtained 0.45 μm and 0.22 μm filter (PVDF) membranes from Merck Display Materials Co., Ltd. , OptiPrep Density Gradient Medium purchasedfrom Guangzhou Fan-si Biotechnology Co., Ltd. , DNase I , BSA , and gelatin from porcine skin purchased from Sigma-Aldrich Trading Co., Ltd. . Furthermore, we used an overspeed centrifuge , HiPure Viral DNA Kit (D3191) purchased from Majorbio , and 10 × SM buffer . Op density gradient solution was made using Optiprep original solution, mixed with 10 × SM buffer (9:1).http://www.moa.gov.cn/gk/yjgl_1/yqfb/201812/t20181223_6165395.htm). The spleen tissue was completely fixed with 10% formalin solution for 72 h. Two formalin fixed pig spleens 1 g each were taking and cut into small pieces with sterile scissors, and then poured into 2 × sucrose -Triton washing solution for washing, 1,000 r/min centrifugation for 15 min, precipitation was beaten evenly with TE Buffer (pH 9.0), adding SDS to the final concentration of 1%, protease K to 200 μg/mL, and kept in water bath at 48°C for 48 h. And 5–10 mL of pre-cooled 1 × SM buffer (SM buffer filtered by 0.22 μm) was added. The spleen was evenly homogenized and placed in liquid nitrogen and a 37°C water bath alternately three times. Spleens were centrifuged at 4°C for 5 min at 1,000 rpm and 12,000 rpm in succession, and the supernatant was filtered using a 0.45 μm filter. Before using the 0.45 μm filter, 1% BSA solution was filtered through the wetting membrane. Then, qPCR was performed on each liquid layer using specific ASFV primers to determine the virus Cq value in each liquid layer. The liquid layer with the low Cq value was selected, each two liquid layers were mixed, and 1 × μSM buffer solution was added to fully mix. The solution was centrifuged at 160,000 g for 1 h, and the supernatant was discarded. According to the amount of precipitation, 100–500 μL 1 × SM buffer solution was added for resuspension (with repeated pipette mixing to avoid violent shock). DNaseI was added according to the manufacturer's instructions, with treatment at 37°C for 1 h. Single- and double-stranded DNA were excised simultaneously to fragment the DNA for library construction. EDTA  was added according to the manufacturer's instructions to terminate the reaction. The HiPure Viral DNA Kit (D3191)  was used to extract viral DNA, and the concentration was measured. Sequencing libraries were generated using NEB Next® Ultra™ DNA Library Prep Kit for Illumina®  following manufacturer's recommendations and index codes were added. Finally, the library was sequenced on an Illumina Novaseq 6,000 and 150 bp paired-end reads were generated. Each sample added 507,333,357 reads.  for sequencing. All the experiments involving the ASF virus were carried out in a biosafety level (BSL)-3 laboratory at South China Agricultural University .Cases were identified from the Ministry of Agriculture and Rural Affairs of Huangpu District, Guangzhou ; (2) Comparison between BLASTX (v2.9.0+) and the virus-NR database isolated from NR (including bacteriophages) was used to screen the comparison results with e ≤ 1 × 10−3; (3) Metagenemark (v3.38) was used to predict the genes, and then HMMSearch (v3.2.1) software was used to compare the protein sequences with the HMM database (VPFS and VFAM), and the comparison results were screened with e ≤ 1 × 10−5.A variety of methods  were used for functional annotation information.The predicted gene protein sequence and UniProtKB/Swiss-Prot database sequence of the virus [ViralZone , revieweThe contig on ASFV was identified and compared according to the final virus sequence for analysis. MAFFT  softwareAfter centrifugation, the products were layered into Eppendorf tubes, which were weighed before use and after adding each liquid layer. The weights of the liquid layers were recorded and the density of each liquid layer was calculated  to assesTo ensure the accuracy of subsequent analysis, SOAPnuke (v2.0.5) software was first used to process the raw data from the machine, and high-quality clean reads were obtained. The quality distribution is shown in To avoid the influence of the host sequence on subsequent analysis, BWA  software was used to compare clean reads with the host database. Sus scrofa was used as the host reference information (accession: NC_010443.5). The comparison results where the length of the comparison was >80% of the total read length were filtered, and then the corresponding sequence was removed. The results showed that 16.5% of clean reads (PE: paired-end) were obtained after host removal, as shown in Asfarviridae, red column in the figure) accounted for 87.12% of the total reads (PE). Asfarviridae had only one ASFV member, so it was not necessary to make annotations at the genus level for the display. The statistical results are shown in To quickly obtain virus composition information in samples, BWA  software was used to compare clean reads to virus reference data (isolated from NT data); the comparison results were filtered if the length of the comparison was >80% of the total read length. Statistical analysis showed that virus reads (PE) accounted for 3.19% of clean reads (PE), as shown in MEGAHIT  software was used to assemble clean data, and BWA (v0.7.17) software was used to compare clean reads with assembly results. The calculated utilization rate of the reads was 75.30%. At the same time, BLAST (v2.9.0+) software was used to align the assembled contigs with the host sequence and to remove the host sequence. The statistical results showed that the maximum number of 400 bp contigs was 3,896 , and thee ≤ 1 × 10−5, and high reliability; (2) Suspected: did not meet the conditions in (1), compare length ≥100 bp, e ≤ 1 × 10−5, and low credibility, requiring in-depth analysis and verification; (3) According to NCBI Taxonomy annotation information, the number of phages and other viruses, as well as the types of RNA and DNA, were counted (The obtained contigs were compared to the virus database (isolated from the NT database) using BLAST (v2.9.0+) software. Comparison and screening criteria were as follows. (1) Confirmative: comparison of similarity ≥80%, compare length ≥500 bp,  counted .Based on the comparison of reference data, there were two defects: (1) Only known viruses could be identified; (2) There were false positives in the results. To reduce false positives and identify unknown viruses, new methods were used to further identify virus sequences by combining multiple databases . The anaThe virus sequences obtained by the two methods were compared, and Venn diagram were drawn. Then, the confirmed and novel virus sequences (Section 3.6.1 and 3.6.2) were combined as the final virus sequences .BWA  software was used to compare the host-removed clean reads with the obtained virus contigs, and the proportion of virus reads was calculated when the filter comparison length was lower than 80% of the total read length. The RPKM value of each viral contig was calculated. The maximum RPKM value was 85,317.57 and the minimum RPKM value was 224.62 .Metagenemark (v3.38) software was used to predict the gene corresponding to virus contigs, and sequences with gene nucleic acid length >150 bp were filtered. The statistical results are presented in https://viralzone.expasy.org/). The best hit screened with e < 1 × 10−3 was compared to obtain the function information of the virus. The results showed that the percentage of genes in the database accounted for 37.14% of the total gene number (Based on the BLASTP (v2.9.0+) software, the gene sequences were compared with those collected from the UniProtKB/SWISS-PROT database (e number . Functioe number . The poln = 12), genotype I ASFV strain (n = 8), genotype II ASFV strain (n = 6), genotype I ASFV strain (n = 2), and genotype VIII, IV, XX, III, V, and XII ASFV strains (each one). Phylogenetic analysis of the entire viral genome was performed using the NJ method in Mega X, and the phylogenetic tree bootstrap value was set to 1,000  method directly uses the genetic material of all viruses in the samples for analysis. First, virus particles are enriched, the genome sequence information is obtained, and then the composition and relative abundance of all viruses are identified. This method is a powerful means for the discovery of new viruses and the early detection and control of virus infection. It is of great use in the study of the origin and evolutionary patterns of viruses, genetic diversity, geographical distribution, and the relationship between viruses and their hosts. In the early stage, we used first-generation sequencing to sequence the whole genome of the Guangdong outbreak ASFV, and submitted the sequence to GenBank . Due to the time-consuming characteristic of first-generation sequencing, we improved the sequencing analysis method based on the macro virus group (virome) method. Previous studies used a density gradient buffer of Percoll, sucrose, and cesium chloride , whereasThe results of data quality control showed that the average base mass at all positions was above 30, indicating high data quality. A total of 1,351,282 reads  were mapped to the entire genome region of the MT496893.1 strain, with an average depth of 1,233. Finally, the MT496893.1 strain was used as a reference for genome correction and assembly, and a 188,035 bp genome was finally obtained. If tissue can be ground and supercentrifuged, the high purity and concentration of virus particles can be theoretically stratified for second-generation sequencing, and virus isolation is not required, which will shorten the time to obtain the results . Using cThe original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.HW and GZ: conceived and designed the experiments. CJ, JJ, and YW: performed the experiments. YCh, YW, ZM, MC, HY, GLi, and LG: sample collection. CJ, CQ, YCh, and GLi: analyzed the data. CJ and HW: contributed to the writing. All authors have read and approved the final manuscript.The Key-Area Research and Development Program of Guangdong Province (2019B020211003 and 2019B020211005). National Natural Science Foundation of China (31941004) and China Agriculture Research System of MOF and MARA.CJ, YW, YCh, GZ, and HW were employed by company Shenzhen Kingkey Smart Agriculture Times Co., Ltd. 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The primary objectives of total hip arthroplasty (THA) include mobility improvement and pain relief; however, the correlation between hip range of motion (ROM) and function remains unclear. We aimed to explore how ROM affects hip functions after THA and compare the responsiveness of each component of the modified Harris Hip Score (mHHS) and Oxford Hip Score (OHS) to preoperative and postoperative ROM.This prospective observational study involved 120 patients who underwent unilateral THA. Univariate regression analyses were performed using the University of California Los Angeles activity score and mHHS and OHS to determine the effects of preoperative and postoperative flex ROM on clinical scores at 12 months. Multivariate regressions were performed to adjust for the confounding effects of patient factors: age, sex, body mass index, and diagnosis.P = .0041) at 12 months; the effect on the OHS socks component was not significant (P = .34). A larger flexion ROM at 12 months was associated with higher scores in the mHHS support , stairs , and socks  components but had no significant effect on any OHS component. The effects of ROM on University of California Los Angeles activity score were limited.A larger preoperative flexion ROM was associated with a higher score in the mHHS socks component (standardized coefficient [SC] = 0.26, A discrepancy was noted in the responsiveness to ROM between the two major measurement tools; this difference might be because mHHS and OHS are surgeon- and patient-administered questionnaires, respectively. This discrepancy also suggests that the patients have higher satisfaction than that assumed by the surgeons. Pain and limited mobility are common symptoms of hip osteoarthritis (OA). Restricted hip range of motion (ROM) increases lower limb disability in patients with hip OA ,2. Thus,Limited studies have been conducted regarding the correlation between hip ROM and function score. Therefore, the correlation between hip ROM and function remains controversial; some studies have reported ROM as a determinate of function ,4, whereHip ROM can have different effects on various clinical outcome measurement systems; however, these also remain unknown. The HHS and Oxford Hip Score (OHS) are the two most frequently used clinical measurement tools. The HHS is a surgeon-administered joint-specific questionnaire comprising 10 items: 2 (ROM and absence of deformity) for the physician physical examination component and 8 for the patient-reported outcome measure (PROM) component. When only the PROM component of the HHS questionnaire is used, it is referred to as modified HHS (mHHS). The OHS is a 12-item self-administered questionnaire. These two measurement tools share similar questions regarding function components  and are considered to be strongly correlated ,10. CorrThis study aimed to explore how the preoperative ROM affects postoperative hip functions after THA and compare the responsiveness of each component of mHHS and OHS to preoperative and postoperative ROM.This was a prospective observational study. All patients provided informed consent, and the study protocol was approved by the institutional review board of our hospital.Between July 2014 and July 2016, 200 primary THAs (in 183 patients) were performed at our institute. Among these, 78 THAs were excluded because the condition of the contralateral hip might have affected the clinical activity scores. Specifically, the contralateral hips of the excluded patients had untreated severe OA (n = 39), a total hip joint implanted within 12 months before the index surgery (n = 24), and untreated osteonecrosis of the femoral head (n = 15). Two more THAs were excluded because the procedure involved subtrochanteric shortening osteotomy. Therefore, the present study cohort included 120 THAs in 120 patients whose contralateral hip was either normal or had undergone THA within the prior year.This cohort was the same group of patients used in another prospective study that aimed to evaluate the changes in the PROM after THA .All 120 procedures were performed by one of the senior hip surgeons . All surgeries were performed via the anterolateral approach [The following patient-related covariates were considered: age, sex, body mass index (BMI), and diagnosis.Clinical scores were evaluated preoperatively and postoperatively at 3, 6, and 12 months using the mHHS, University of California Los Angeles activity score (UCLAAS), and OHS. All patients were evaluated using all three scores. The standard HHS comprises 10 questions: two (ROM and absence of contracture) for the physical examination component and eight for the PROM component. In this study, the mHHS comprised only the eight PROM components but not the ROM and contracture components included in the standard HHS.The OHS was calculated in accordance with the modified scoring system; each question was scored from 0 to 4, with 4 representing the best outcome or the least symptoms. The OHS ranged from 0 to 48, with 48 indicating the best outcome. One of the surgeons filled out the mHHS questionnaire by asking each question to the patients; the UCLAAS questionnaire was also filled out by a surgeon in a similar manner. The OHS questionnaire was handed out to the patients; it was collected by the hospital staff after the patients had completed the questionnaire.The hip flexion ROM was measured preoperatively and postoperatively at 3, 6, and 12 months after THA by one of the senior orthopedic surgeons through visual estimation. Previous studies have revealed that the intraclass correlation coefficients (ICCs) for the interobserver reliability and intraobserver reproducibility of hip ROM measurements obtained through visual estimation are the highest for hip flexion ,14. In tDifferences in proportions were analyzed using the chi-square test. Differences in means were analyzed using the Wilcoxon test for the comparison of two groups. Univariate regression analyses were performed to determine the effects of preoperative and postoperative flex ROM on clinical scores  at 12 months. Multivariate regressions were performed to adjust the confounding effects of patient factors: age, sex, BMI, and diagnosis (OA or others). Probability values of <0.05 were considered significant. The sample size required for between-group comparison was calculated using bilateral alpha risk of 5%, 90% power, standard deviation, and anticipated differences for each function score. At least 97 patients were required. With an anticipated exclusion rate of 40%, we included 183 patients for analyses. All statistical analyses were performed using the JMP Pro 14 software . The correlations between clinical measurement scores  and ROM at each time point were also visualized using the nonparametric Kernel density estimation using a standard deviation multiplied by 0.225 (the default setting for JMP Pro 14) for each variable as the Kernel standard deviation for each variable. Quantile density contours were drawn at every 0.05.P = .073).Patient demographics are presented in P = .018) but not preoperatively or at 6 and 12 months. The total mHHS was significantly associated with flexion ROM at 6 and 12 months . At 3 months, total mHHS showed a tendency to be higher when flexion ROM was larger, but this correlation was not significant (P = .074).Correlations between the clinical scores  and flexion ROM at each time point until 12 months of surgery are illustrated in P = .07) and 6 (P = .063) months. The established value for minimally clinically important difference (MCID) are a difference of 8 points for mHHS [The total score and scores of each component in OHS, mHHS, and UCLAAS for all patients in the small and large preoperative ROM groups are shown in for mHHS , 5 pointfor mHHS ,17, and for mHHS . Any of P = .045) and socks (P < .001) components. At 12 months, there was a significant difference only in the socks component (P = .0071).Preoperatively, the large preoperative ROM group had higher scores in the mHHS limp . No differences were noted between the two groups in other component scores preoperatively or at 12 months.The large preoperative ROM group had a higher socks component score preoperatively and at 12 months although the difference was not statistically significant (P = .071). Any of the difference in those three measurements between small and large postoperative ROM groups did not reach the MCID.The total score and scores of each component in OHS, mHHS, and UCLAAS for both groups are shown in P = .031), stairs (P = .011), and socks (P < .0001) components at 12 months.The large ROM group had higher scores in the mHHS limp (There was no significant difference in OHS at 12 months between the two groups in any component.P = .99).In the socks component at 12 months, the scores for both ROM groups were close ; however, this effect was not significant on the OHS socks component (P = .34).The effects of preoperative flexion ROM on the total score and scores of each component at 12 months after surgery are shown in P = .026), stairs , and socks  components. At 12 months, flex ROM had no significant effect on any OHS component.The effects of 12-month postoperative flexion ROM on the total score and scores of each component at 12 months after surgery are shown in The mean flexion ROM increased from 80.9° (range: 10°–110°) preoperatively to 95.0° (range: 40°–110°) after 12 months of THA. Despite this improvement, the mean postoperative ROM did not reach normal levels. This may be because the soft tissues have a major impact on impingement-free ROM after THA ,20. For P = .041). However, the postoperative score in the OHS socks component was not affected by preoperative flex ROM.The present study evaluated the effects of preoperative and postoperative flexion ROM on mHHS, OHS, and UCLAAS. Even after adjustments for confounding factors such as age, BMI, sex, and diagnosis, the preoperative ROM was associated only with the postoperative score in the mHHS socks component .The effects of flex ROM at each time point on total mHHS and OHS were limited . There wP = .026), stairs , and socks components , whereas there was no significant effect on any OHS component. However, the difference in component score in mHHS between small and large postoperative groups was small (0 to 0.83; At 12 months, the effects of ROM were significant in the mHHS support (SC = 0.21, OHS is a PROM questionnaire that patients fill out by themselves, whereas HHS is scored by physicians. OHS and HHS are known to be strongly correlated ,10. HoweKenneth et al. reported that a larger postoperative hip ROM was associated with a higher total mHHS . HoweverTo the best of our knowledge, no reports have described the correlation between the UCLAAS and hip ROM. The present study found no correlation between the UCLAAS and postoperative hip flexion ROM at 12 months . This coOlder age has been associated with a smaller ROM . Loss ofQuality assessment using PROMs is becoming more important in the field of orthopedics ,31. VariThe limitations of this study include the relatively small number of patients and the heterogeneity in diagnosis and the use of cement. Another limitation is the lack of information on the condition of the lumbar spine. A stiff spine would make various activities, including wearing socks, difficult even with a large hip ROM. The difference in each component score between small and large ROM groups was not compared with MCID because of the lack of established MCID for component score in mHHS and OHS. This would be another limitation for this study.In conclusion, smaller postoperative flexion ROM was associated with lower scores in the mHHS support, stairs, and socks components after THA even after adjustment for other patient-related confounders. There were no significant associations between ROM and these component scores in the OHS. This discrepancy in responsiveness to ROM between the two measurement tools may be because the HHS uses a surgeon-administered questionnaire, whereas the OHS uses a patient-administered one. The discrepancy also indicates that patients actually have higher satisfaction than that assumed by the surgeons. Surgeons should be aware that patients may not feel as much difficulty because of small ROM as they assume.S.M. receives research grant from Kyocera and Smith & Nephew; the other authors declare no potential conflicts of interest.