The resulting mesoporous silica can be used for removing/capture of CO2 from emissions or for wastewater treatment. The synthesis of various porous materials using wastes would allow a high level of recycling for a sustainable society with low environmental impact.In order to meet the increasing energy demand and to decrease the dependency on coal, environmentally friendly methods for fly ash utilization are required. In this respect, the priority is to identify the fly ash properties and to consider its potential as raw material in the obtaining of high-value materials. The physico-chemical and structural characteristics of the fly ash coming from various worldwide power plants are briefly presented. The fly ash was sampled from power plants where the combustion of lignite and hard coal in pulverized-fuel boilers (PC) and circulating fluidized bed (CFB) boilers was applied. The fly ash has high silica content. Due to this, the fly ash can be considered a potential raw material for the synthesis of nanoporous materials, such as zeolites or mesoporous silica. The samples with the highest content of SiO Global coal consumption rose with 0.9% in 2018, the main contributors being India and China, followed by Turkey and Russia [By contrast, the United States reached the lowest level in the last 40 years, the coal consumption decreasing 4% in 2018 due to the availability of natural gas at lower prices and to stronger emission legislation .2 emissions are strongly related to coal consumption, and consequently the United States was expected to reduce CO2 emissions by 2.2% in 2019 by further reducing coal consumption, and by 3.6% in 2020 [CO2 emissions costs have been responsible for a decrease of coal consumption in the last 6 years in Europe  ,9, Y (FA2/g) and good stability in thermal conditions [The mesoporous silica materials, known since 1992 as M41S, present great potential in worldwide applications, such as catalysis or wastewater treatment, due to their properties, namely, uniformity of pore distribution (with size between 2 and 50 nm), high surface area,  is significantly different. Moreover, the synthesis conditions, including the pre-treatment step, are often not completely described or some inconsistencies among the literature sources were observed.This overview systematically explores the synthesis of various mesoporous silica materials derived from fly ash by taking into consideration the waste properties (fly ash), pre-treatment procedures or the hydrothermal treatment parameters .2 > Al2O3 > Fe2O3 > CaO > MgO > K2O and large amount of unburned carbon. Furthermore, fly ash contains trace elements that can have a negative impact on the environment [Fly ash is a complex material, being a by-product resulting from the combustion of various coals with high contents of minerals ,21,22,23ironment ,25,26. Tironment . Also, w2 contents were higher in the fly ash derived from sub-bituminous (40%–60%) and bituminous (20%–60%) samples than in lignite samples (15%–45%). The same trend was seen for Al2O3. In the case of CaO and MgO higher contents were observed in fly ash from lignite samples , followed by sub-bituminous samples.As it is shown in 3, Na2O and K2O were observed in all the fly ash samples. The highest percent value for unburned carbon, determined by its loss-on-ignition (LOI), was found in bituminous fly ash (15%).Small contents of SO2 and Al2O3, their recovery being an issue raised by the waste management. Also, the high content of CaO can be used for CO2 capture and permanent sequestration, resulting in CaCO3 [Comparing the sample contents, it can be highlighted that all types of fly ash are rich sources of SiOin CaCO3 . Bitumin2 capture [The intensive investigations carried out for fly ash reuse have resulted in the development of techniques for producing glass ceramics, ceramic wares, silicon carbide, silicon nitride, hollow/masonry/concrete blocks, cordierite or mullite. Recently, fly ash was applied in the development of mesoporous silica for CO capture .The reuse of fly ash must be encouraged for many reasons. For example, the disposal costs would be minimized; also, less landscape would be reserved for its disposal; and the by-products may be used as raw materials.Fly ash may have metal concentrations up to 10 times higher than coal . As it wA typical metallic composition in fly ash collected from different power plants in India is presented in As it can be seen from Regarding the concentration of toxic elements in soil and crops, It was observed that Fe was the major element. As it can be seen in It was also demonstrated that the soil treated with fly ash contributed to the increase of crop yields, by the action of essential plant nutrients, such as Mg, S, K, Ca, Mn, Fe and Zn . The flyAs it can be seen, the use of coal fly ash meets technical, economic and legal barriers. The main technical milestones refer to coal fly ash production, specifications and standards, product demonstration and commercialization.For coal-using plants, the incomes from the sale of fly ash are often negligible, the most important economic barriers coming from the increased cost of transportation of fly ash and competition from locally available natural sources. Legal barriers resulting from the lack of knowledge regarding the potential ash application, insufficient data on environmental and health risks, lack of regulations and procurement guidelines.Worried industry and government representatives, scientists, and engineers have created national and international organizations to overcome the milestones of coal fly ash reuse. An integrated approach must be designed in order to produce a superior quality of fly ash-based materials, which can satisfy the consumer’s expectations. Furthermore, high quantities of fly ash are often followed by emission of greenhouse gases, significantly intruding global warming. As a consequence, to conform to the environmental requirements, major efforts must be achieved in fly ash management, reducing the negative effect on the environment.A deep analysis on the cost of fly ash use versus conventional building materials is required, being necessary to apply the best engineering practices in order to minimize as much as possible the environmental risk.In order to increase the recycling of the coal fly ash and to reduce the environmental risk of contamination with heavy metals, several applications for recovery of valuable components from fly ash were developed.2 reduction and wastewater treatment.Thus, among the fly ash used in conventional applications (cements obtaining), various mesoporous silica materials with improved properties were obtained, able to be applied in environmental depollution, namely COThe methods for obtaining these materials from fly ash were basically similar, the differences consisting in varying different parameters such as: pH, the temperature and the time of hydrothermal or aging treatment and also the quantities of raw materials or reagents. These variations influenced more or less the obtained material properties.The used coal fly ash for the silica synthesis had various particle sizes (2.0–30 µm) and chemical compositions  ,36,37,382/g, pore volume: 0.75 and 0.85 cm3/g and pore diameter: 3.7 and 2.7 nm) it was observed that the Al-MCM-41 from the conventional method was more efficient in the cumene conversion because not all of the Al presented in the fly ash-derived porous material had catalytic activity. Thus, the cumene conversion was threefold higher, ~22%, at the beginning of the reaction for the Al-MCM-41 prepared from pure reagents and decreased drastically during the first hour, slowly decreasing the next hour for the two materials, reaching 6%, two times higher than Al-MCM-41 derived coal fly ash.Porous materials, such as Al-MCM-41 and SBA-15, were obtained from coal fly ash, and they were used as catalysts in the cumene cracking reaction . In this3CN, Toluene, Tetrahydrofuran (THF), CH2Cl2 and H2O). A 90% yield was obtained in 4 h for 0.2 g MCM-41 in ethanol, providing a good catalytic active area for the reaction. The yield remained constant with the increase of catalyst quantity at 0.3 g. Instead, a lower efficiency, down to 50%, was observed with the decrease of catalyst quantity, to 0.1 g, even if the reaction time has been extended to 8 h [MCM-41 was also obtained by adapting the mesoporous synthesis from the previous study , with mod to 8 h . The MCM2/g, the pore diameter decreased from 7.54 to 5.13 nm and the pore volume increased from 0.13 to 0.71 cm3/g, although not reaching the properties of the material obtained through conventional methods [A supernatant with higher contents of Si, Al and Na  was obtained starting also from the method previously described , the fly methods . The Al-2 and Al2O3 ranging from 46.15 wt% to 56.52 wt% and from 18.48 wt% to 31.06 wt%, respectively. The lowest and the highest value of SiO2 were obtained from PC combustion. Compared to other studies [MCM-41 was also synthesized from brown and hard coal fly ash (10 samples) using two procedures, namely pulverized-fuel (PC) and fluidized-bed boilers (CFB) . The X-r studies ,37, wherIt was observed that the impurities presented in the coal fly ash composition, such as Fe, Ca, K, S and P, were also present in the channels of the obtained mesoporous material, conducting to poor properties compared to the commercial material .For comparison, the ash resulting from the rice husk combustion was tested, using similar extraction steps as in the case of fly ash . Even ifIt was concluded that, despite the similar chemical composition, the textural properties of the rice husk and fly ash-derived materials were superior to the commercial MCM-41, presenting higher pore volume for all synthesized materials, higher specific surface area and similar morphologies.2 capture. The results shown higher CO2 uptakes (with 2 wt.%) for the synthesized materials containing 60 wt.% PEI, the materials having faster kinetics due to larger pore volume comparing to the commercial materials.Also, the reaction time influenced the obtained suspension containing the surfactant (hexadecyltrimethylammonium bromide-CTAB) . A longeIt was established that the fly ash desilication rate can reach 46.3%, by using sodium hydroxide at a mass ratio of 1:6.4, during 4 h under thermal condition (95 °C). This solution was used as silica source to obtain SBA-15 mesoporous silica. The method involved further hydrothermal treatment for two days at 110 °C and a triblock copolymer as template-poly(ethylene oxide)/poly(propylene oxide)/poly(ethylene oxide) .2- groups, and toluene as solvent [2+ adsorption [The functionalization of SBA-15 with amino groups involved, as in case of MCM-41, aminopropyltriethoxysilane (APTES), as source of NH solvent ,43. An a2+ initial concentration on amino-SBA-15 adsorbent was studied and it was remarked that the adsorption equilibrium time was achieved after 1 h, reaching 131 mg/g  and the pore volume (~0.96 cm3/g) increased with the increase of Si/Al molar ratio, up to 25. The efficiency of the aluminosilicate on the methylene blue and crystal violet removal from wastewater was evaluated, revealing its high adsorption capacity  from wastewater. In this respect, mesoporous aluminosilicate was synthesized from fly ash by varying the Si/Al molar ratio . X-ray dcapacity . Higher Commercial MCM-41 production is expensive, due to intense energy consumption or utilization of inorganic and organic silicate reagents, the identification of new cheap silica sources being a viable future solution ,45. BesiThe synthesis of silica from waste is not free of cost, because the procedure is similar to the conventional methods, involving hydrothermal treatment with intensive energy consumption. The difference between conventional and unconventional synthesis is the source of silica, which leads to extra synthesis steps. The significant reduction of the synthesis time, from 72 h  to 2 h, could result in the cost decrease, by saving energy . The estTaking into account the depollution and storage costs for fly ash or other silica source wastes, it could lead to a fair price and a sustainable process.In the last few decades, worldwide coal consumption has been monitored. High quantities of fly ash are discharged as a by-product from the coal power plants. The influence of fly ash on soil-crops chain (maize and rice) in different areas was demonstrated. In this respect, an effective process to recycle this waste is mandatory.Fly ash proved to be suitable to use in environmental applications, for example by replacing activated carbon and zeolites as adsorbents for air pollutants or wastewater treatment. Its adsorption capacity mainly depends on the origin and the activation method. To date, no industrial set-up has been developed. Economic milestones must be surpassed. Taking into consideration the amorphous alumina-silicate nature of fly ash, it can be applied as raw material in various industrial reactions, such as obtaining ultramarine blue. Intensive harvesting depletes trace elements in the soil. Despite the fact that many studies were achieved for the use of fly ash as soil amendment, full-scale application has not been accomplished. In the near future, farmers may consider fly ash to be a substitute for lime to enrich the soil.2 from gaseous emissions or for various water pollutants. The synthesis of different nanoporous materials using as raw materials various wastes would enable a high level of recycling for a sustainable society with a low environmental impact.Fly ash has high silica content, which makes it a potential source for obtaining nanoporous materials, such as mesoporous silica. In this overview, various methods for obtaining mesoporous silica materials from fly ash were examined by considering the ash composition, pretreatment procedures, and synthesis conditions, in order to identify the optimal parameters for synthesis. The physical properties of the materials derived from fly ash confirmed their mesoporous nature, similar to the mesoporous silica materials obtained by conventional methods. The mesoporous silica materials with a high surface area and large pore volume derived from fly ash can be applied as a support or as a surface-functionalized host for capturing CO


In the cluster analysis, we categorized the patients into three groups: no adipose tissue accumulation (NAT group), SAT prior accumulation (SAT group), and visceral adipose tissue prior accumulation (VAT group). The results of the three groups revealed that the SAT group displayed a significantly poor prognosis of the Kaplan–Meier curve  of a 5-year survival rate. Propensity score matching analysis of the SAT and VAT groups was performed to adjust the patient’s background, but no significant differences were found between them; however, the prognosis was poorer , and hemostatic factors were still at a lower level in the SAT group. These findings suggest that SAT accumulation type of obesity is a poor prognostic factor in alcoholic LC patients without HCC, and the hemorrhagic tendency might worsen the poor prognosis in such cases.In alcoholic liver cirrhosis (LC) patients, obesity has become a problem that progresses into liver dysfunction. Herein, we investigated the relationship between the prognosis of steatohepatitis and body weight, along with fat accumulation in patients with alcoholic LC. We conducted a single-center retrospective study, enrolled 104 alcoholic LC patients without hepatocellular carcinoma (HCC) based on histological and clinical evidence, and investigated factors related to poor prognosis using multivariate Cox regression and cluster analyses. Cox regression analysis revealed three independent relevant factors: subcutaneous adipose tissue (SAT) index (median 34.8 cm Because of the development of antiviral agents with negligible adverse effects switched from interferon therapies, liver diseases caused by the hepatitis C virus have decreased gradually and undoubtedly . TherefoThe distinction between alcoholic and nonalcoholic liver diseases is often difficult, owing to the unreliability of alcohol consumption history . When chPrevious studies showed that nutritional disorder that focused on malnutrition and vitamin deficiency accounted for patients with alcoholic liver cirrhosis (LC), and heavy drinking, irregular feeding habits, and decompensated cirrhosis with ascites are involved in malnutrition . In factHerein, we investigated and reported the relationship between prognosis and body weight, as well as fat accumulation in patients with alcoholic LC.This retrospective study was performed at Niigata University Hospital. The study was approved by the ethical review board of Niigata University .Data were collected from hospital medical records of patients diagnosed with LC between January 2006 and December 2019. LC diagnosis was based on histological and clinical evidence. Clinical diagnosis of LC was based on a previous report , as follBecause of Niigata University Hospital is the higher-level functional hospital in Niigata prefecture, Japan, patients were referred from other hospitals for professional treatment, such as endoscopic submucosal dissection for esophageal cancer, endoscopic injection sclerotherapy or variceal ligation, or balloon-occluded retrograde transvenous obliteration of gastroesophageal varices, and surgery or transcatheter arterial chemoembolization for HCC (HCC cases were excluded in this study). Therefore, we performed CT in almost all cases (approximately 90%) to determine the therapeutic strategy for these problems.A transverse CT image from the third lumbar vertebra was identified from each retrieved scan. The CT images were analyzed using hand segmentation with the help of image analysis software SliceOmatic version 5.0 .2) and expressed as SMI, SAT index, and visceral adipose tissue (VAT) index in cm2/m2. The total adipose tissue (TAT) was determined by the total of the SAT and VAT. Besides, TAT was also normalized for stature .The identification of skeletal muscle and adipose tissue was based on predefined CT density of −29 to +150  and −190Cox regression and Kaplan–Meier analysis were used to compare the prognosis, and the cumulative incidence plots method was used to compare the plausible incidence of HCC. The Kolmogorov–Smirnov test was used to assess the normality of the distribution of continuous variables. Spearman’s rank coefficient of correlation was used to evaluate correlativity. Mann–Whitney U, Kruskal–Wallis, and Fisher’s exact tests were used to compare data of each cluster. The propensity score matching analysis was used to adjust the factors of SAT and VAT patient groups. Propensity score was calculated using the adjusting factors: age, body mass index (BMI) and total bilirubin level. Cases in the VAT group were selected that corresponded to those in the SAT group by the nearest neighbor method. SPSS Statistics software  was used to perform Spearman’s rank coefficient of correlation, the Kolmogorov–Smirnov, Mann–Whitney U, Kruskal–Wallis, and Fisher’s exact tests, and propensity score matching, whereas the Prism Software  was used to perform Kaplan–Meier and the cumulative incidence plots.s = 0.669), VAT index (rs = 0.484), and TAT index (rs = 0.635). 45 cm2/m2 of the SAT and VAT indexes and 90 cm2/m2 of the TAT index were equivalent to 25 kg/m2 of BMI, and 55 cm2/m2 of the TAT index was equivalent to 20 kg/m2 of BMI in the correlation analysis , with a median age of 60 years. The median observation period in the entire cohort was 1.6 years. Furthermore, 16 patients 15.4%) had HCC as a complication, and 30 (28.8%) died during the study period. The median score of the Charlson comorbidity index [.4% had Hanalysis .2/m2, 5-year survival rates were 66.5 vs 22.3, 2, 5-year survival rates were 67.7 vs 25.1, First, we investigated the relevant factors for prognosis in 104 patients with alcoholic LC by univariate Cox regression analysis  and foun2, 5-year incidence rates were 25.5 vs 49.5, 2/m2, 5-year incidence rate were 31.5 vs 37.0, 2 of BMI. In addition, one-year survival rate was 87.1, and 3-year survival rate was 46.4 after the incidence of HCC , and multivariate Cox regression analysis revealed only two factors, Charlson comorbidity index , and BMI , which remained significantly different . The cume of HCC .2/m2, NAT group), SAT prior accumulation , and VAT prior accumulation  as the cluster analysis , BMI , total bilirubin , and prothrombin time , fibrinogen , and platelet count . Therefore, we compared two groups using propensity score matching analysis after adjusting for the three factors, age, BMI, and serum total bilirubin level. Twenty-seven SAT, and VAT patient groups were 1:1 matched, and no significant differences were found between the two groups in terms of age, gender, BMI, and the ALBI grade . The pro3) were at greater risk for HCC (4.8% a year). In our study, the median age was 60 and median platelet count was 93,000/mm3, which indicated that patients at high risk for HCC were included more than in previous studies. Furthermore, Ganne-Carrié et al. [Obesity is the risk factor for alcoholic LC as previously mentioned . The evié et al.  indicateSince the condition of many LC patients were complicated with ascites that might cause inaccuracy of the BMI measurements, we used L3-level adipose accumulation in CT scan to exclude the influence of ascites and revealed SAT accumulation was more marked than BMI for prognosis in alcoholic patients with LC.2 of VAT area [It is well known that VAT accumulation generates a harmful effect on the human body. A large clinical study in Japan revealed that the mean number of obesity-related cardiovascular risk factors  significantly correlated with the VAT area but not the SAT area . In the VAT area  by the mThere is no precise elucidation regarding the difference in the distribution and function between SAT and VAT; however, it is known that the SAT tends to exhibit differential proliferation but not VAT . For hypAlthough clinical results and mechanisms have been established previously, our data revealed that the accumulation of SAT significantly correlated with poor prognosis. According to our analysis of the cause of the death in alcoholic LC, a significantly high incidence of fatal bleeding complications was found, primarily, non-variceal bleeding, as typified by muscle hematoma. On the other hand, no significant difference between liver failure and HCC was noted. Alcoholic LC had a higher incidence of non-variceal bleedings than other occlusions of cirrhosis , and theOur present study has the following limitations. First, the amount of alcohol intake after the diagnosis of alcoholic LC could not be assessed because this is a retrospective study. It is well known that complete abstinence from alcohol could reduce the risk of liver-related complications and mortality , 35, andSAT accumulation-type obesity in alcoholic LC patients without HCC is significantly related to poor prognosis. In this type, the complication of non-variceal bleeding, including muscle hematoma, characteristics in alcoholic LC was found. To follow these cases, we should pay more attention to the complication of HCC and bleeding. To predict the bleeding risk, we should confirm the hemostatic and coagulation factors and the adipose distribution.S1 FigOne-year survival rate was 87.1 and 3-year survival rate was 46.4.(TIF)Click here for additional data file.


ContiThe continuous progress achieved in terms of the cell size and relative density decrease obtained during these years raised the expectations regarding these advanced materials, aiming to reach even smaller cell sizes and larger porosities. Moreover, from theoretical predictions and the previous experience of the positive effect of the cell size decrease to the micrometric range on the physical properties of these materials, the nanocellular polymer foams were expected to present exceptional physical properties. For instance, nanocellular foams could have improved thermal insulation, low dielectric constant, enhanced mechanical performance, or even optical transparency ,3. This Accordingly, as the nanocellular foams field has continued to grow since 2015, it is necessary to perform a critical analysis of its progress, evaluating the fulfilment of the created expectations, the advances in its understanding, the resolved and ongoing challenges, and the key focus points through which to maintain the growth of this field. Herein, this analysis is structured as follows. First, the latest advances in the production of nanocellular foams are briefly summarized, highlighting the most relevant achievements and challenges. Then, the progress in the study of their physical properties is discussed, followed by an evaluation of the success in overcoming the challenges mentioned above. Finally, the main ideas extracted from this critical analysis are presented, identifying the most challenging, and promising, current focus points in this field, as well as potential applications identified in the last few years, which is the main aim of this Special Issue.15 cells/cm3 [Decreasing the cell size is, without doubt, the motivation behind research on nanocellular foams. In the last few years, several works have achieved the production of nanocellular foams with cell sizes below 100 nm. On the one hand, some crystalline polymers have finally provided such features, but showing severe limitations in terms of density reduction, as cell nucleation and growth can only occur following the pattern of the amorphous phase between crystalline lamellae . Some ofells/cm3 ,7. Neverells/cm3 ,8,9.15 nuclei/cm3, and relative densities between 0.65 and 0.75. Meanwhile, Guo et al. [On the other hand, several amorphous polymers have shown the capability of producing nanocellular foams , acrylonitrile–butadiene–styrene terpolymers, poly(aryl ether ketone), and thermoplastic polyurethane) ,13,14,15o et al.  achieved2 solubility on the PMMA, generally by increasing the saturation pressure or decreasing the saturation temperature, as well as the optimization of other foaming parameters  and the polymer matrix chemistry are the available tools for researchers working on the homogeneous nucleation route. Ono et al. [15 nuclei/cm3, and relative densities down to 0.44. Yeh et al. [w = 81 kg/mol) are more suitable to decrease the cell size, although they present more difficulties for density reduction. In particular, using a saturation temperature of 0 °C and pressure of 13.8 MPa (CO2 solubility of around 38 wt.%) and foaming temperatures of around 80 °C, they reached cell sizes of around 37 nm, cell densities of around 1016 cells/cm3, and relative densities of 0.25. The same conclusions were achieved by Martin-de-Leon et al. [2 solubility up to 45 wt.%), and foaming temperatures up to 80 °C to obtain cell sizes of around 75 nm, cell nucleation densities of 1015 nuclei/cm3, and relative density of 0.24 for the low-viscosity PMMA grade employed , as well as cell sizes around 14 nm, cell nucleation densities of 3.5 × 1016 nuclei/cm3, and relative density of 0.4 for the high-viscosity PMMA grade .The increase of the COo et al.  employedh et al.  studied n et al.  with a s2 nanoparticles (average size 15 nm) on a PMMA with high molecular weight (Mw = 81 kg/mol) and foamed the samples at a saturation temperature of 0 °C, saturation pressure of 13.8 MPa, and foaming temperatures of around 80 °C, achieving inhomogeneous cellular structures with nanocellular regions with cell sizes of around 70 nm and cell densities around 3.5 × 1014 cells/cm3. It should be noted that, in this case, the introduction of the nanoparticles negatively influenced the cellular structure, as better results achieved by homogeneous nucleation under the same foaming conditions were previously mentioned. Shi et al. [2 appears on the samples during the foaming process. These well-dispersed microcrystals were proven to act as nucleating agents, achieving cell sizes of 87 nm and relative densities of 0.86 by foaming PMMA/PVDF blends with 40 wt.% of PVDF, with the small cell size and high relative density being related to the rigidity of the semicrystalline phase. Although the results obtained by the heterogeneous nucleation route in the last few years have not been quite successful in achieving cell sizes below 100 nm, a few additional works are worth mentioning due to their contribution to the understanding and optimization of this approach. Liu et al. [2-philicity, enhances the cell nucleation efficiency of the nanoparticles from 0.04 to a remarkable value of 0.5. Moreover, they found that modified nanoparticles with sizes of around 80 nm present higher efficiencies than nanoparticles with sizes of around 40 nm and explain this behavior in terms of the line tension effects on the nucleation process. Interestingly, they propose that the same theoretical approach could be applied to justify the nucleation efficiency near 1 previously found in PMMA/MAM blends [On the contrary, the heterogeneous nucleation route offers additional tools to the researchers, related to the nature, size, distribution, and physical properties of the additional phases introduced, their proper distribution being the main challenge of this route. Only a couple of works have been able to achieve cell sizes below 100 nm following this approach. Yeh et al.  introduci et al.  producedu et al.  proved tM blends . Needle-M blends , findingM blends . In part2 solubilities, or even to improve the followed process [2 solubility using higher pressures or lower temperatures involves increasing production costs).The promising results briefly summarized above clearly show that the objective of cell sizes below 100 nm and low relative densities has still not been achieved. The gas dissolution foaming approach still has potential to induce higher CO process , while fHowever, simple geometrical considerations and the physical dimensions of the polymer chains are enough to demonstrate that cell sizes equal to or below 10 nm and low densities cannot be achieved with close-cell geometries . On the The expectations of achieving enhanced physical properties by the production of nanocellular polymers were the main motivation behind the development of these materials. In the last few years, significant knowledge about the mechanical, optical, and thermal properties of these materials has been developed. Wang et al. ,30 have The potential optical transparency of nanocellular foams obtained from transparent amorphous polymers was a controversial topic for some time . HoweverThe thermal conductivity of nanocellular foams is one of its most promising physical properties, as the presence of the Knudsen effect, experimentally validated in nanocellular polymers in 2015 , could lg) of the amorphous polymer matrix. The increase in the Tg has been widely reported on PMMA nanocellular foams [Another thermal property which suffers changes in nanocellular polymer foams is the glass transition temperature  exploring the use of dispersed non-nanostructured block copolymers, but mostly focusing on the development of open-cell nanocellular foams. Second, there is still room for the understanding and optimization of the physical properties of these materials. New approaches to decrease the radiation term on the thermal conductivity of nanocellular foams are urgently required, while the potential of bimodal cellular structures should be further explored. Moreover, the production of transparent nanocellular foams requires additional progress to identify a compromise between their transparency and their thermal insulation. In addition, as a key aspect in improving the understanding of these materials and their physical properties, additional work is required in the study of the increased Tg and confinement effect of nanocellular polymers, using both experimental and theoretical approaches employing MD or similar techniques. Third, the development of strategies to effectively remove the solid outer skins, or, even better, to avoid their formation, as well as continuous processes capable of producing nanocellular foams, are essential for several potential applications and the future industrialization of these materials.Accordingly, several current main challenges in the advanced nanocellular polymer foams field have been identified. First, the simultaneous reduction of the cell size and density requires further technical developments (to increase the COAdditionally, new applications should be considered and tested for these materials. For instance, PMMA-based materials have shown the capability of treating polluted water ,42,43, a


Hydatid disease (HD) is a zoonotic parasitic disease caused by the larvae of Echinococcus, It is mainly prevalent in pastoral areas. Bone echinococcosis is rare, accounting for 0.5 to 4.0% of all echinococcosis. It is likely to miss the diagnosis and misdiagnose due to non-specific early symptoms and the clinical manifestations and imaging features. The clinical data of 15 patients with pelvic cystic echinococcosis were analyzed retrospectively, and the X-ray, CT, and MRI imaging features of the disease were discussed, which are reported below.All 15 patients underwent CT scan evaluation. A total of 8 patients underwent coronal, sagittal, and three-dimensional reconstruction with 3-mm-slice thickness, and 4 patients underwent X-ray plain film examination. Five cases underwent MRI scan. Eight cases underwent MRI or CT enhanced scan.X-ray plain film is characterized by continuous cystic bone destruction, irregular low-density shadow when invading soft tissue, and sometimes calcification which can be seen on the wall or inside the cyst. The involved sacroiliac joint or hip joint may narrow or disappear. The involvement of pelvic cystic echinococcosis is relatively wide, and 80% of patients with pelvic cysts in this group had multiple lesions in the same period. Cystic expansive bone destruction was the most common. Pelvic CT revealed a lobulated hypodense lesion of varying size with internal septae, causing cortical thinning and destruction. Most of them had no periosteal reaction. The iliopsoas muscle is most easily invaded. Single cystic echinococcosis of pelvis showed intermediate or low signal intensity on T1-weighted images and hyperintensity on T2-weighted images in the involved bone and surrounding soft tissue on MRI, and the cyst wall showed linear low signal in T1WI, T2WI, and STIR sequences. The polycystic type is characterized by multiple cysts of varying signal intensity (daughter cysts) within a larger cyst is the typical MRI finding, forming “small vesicles” high signal daughter cysts. Osteosclerosis or calcification showed low signal in T1WI and T2WI.The results of this study suggest that the lesions of pelvic cystic echinococcosis are mostly cystic expansive and osteolytic bone destruction, which is easy to invade the surrounding soft tissue, often accompanied with calcification; among them, multiple cystic lesions are characteristic. Hydatid disease (HD) is a zoonotic parasitic disease caused by the larvae of Echinococcus, also known as echinococcosis. It is mainly prevalent in pastoral areas, especially in Xinjiang, Qinghai, Ningxia, Gansu, Inner Mongolia, and Tibet. Bone echinococcosis is rare, accounting for 0.5 to 4.0% of all echinococcosis , 2. It iAltogether, 15 patients with pelvic cystic echinococcosis were confirmed by histopathological examination or clinical and imaging examination in the first affiliated hospital of Xinjiang Medical University from February 2011 to June 2019, including 10 males and 5 females, aged 19–58 (37.7 ± 9.8) years. There were 4 cases of Kazak and 4 cases of Han nationality, 2 cases of Uygur, Hui and Mongolian each, and 1 case of Kirgiz. Fourteen cases had lived in Xinjiang for a long time, and one case had lived in Qinghai for a long time, with a course of 1–20 years. Clinical manifestations are the following: pain in the hip, lower limbs, and lumbosacral region in 5 cases and limitation of movement in 5 cases (including 1 case with nerve compression symptoms such as incontinence). Local infection occurred in 7 cases . There were 1 case of chronic osteomyelitis, 3 cases of pathological fracture, and no symptoms in 2 cases. There were 7 cases of echinococcosis with other systems, 4 cases of pulmonary echinococcosis, 4 cases of hepatic echinococcosis, and 1 case of prostate echinococcosis. Among them, there were 7 cases confirmed according to the clinical history of echinococcosis combined with laboratory examination and imaging findings, and 8 cases were confirmed by histopathological examination.To record the imaging data of the patients according to the medical records, all 15 patients underwent CT scan evaluation (GE 16- or 64-slice CT machine). A total of 8 patients underwent coronal, sagittal, and three-dimensional reconstruction with 3-mm-slice thickness, and 4 patients underwent X-ray plain film (Philips DR system) examination. Five cases underwent MRI scan (Siemens 1.5T Avanto MR scanner). The scanning sequence included T1WI, T2WI, and short time inversion recovery (STIR); 8 cases underwent MRI or CT enhanced scan , with iohexol (300 mg I/ml), 1.5 ml/kg, and injection flow rate of 3.0 ml/s; MRI enhanced contrast agent was GD DTPA, 0.2 ml/kg, injection flow rate 2.0 ml/s.In 15 cases, sacrum, ilium, pubis, ischia, acetabulum, sacroiliac joint, hip joint, and soft tissue of surrounding muscles were the main affected parts. There were 4 cases of echinococcosis in the same side of femur and 3 cases of echinococcosis in the lumbar and sacral vertebra. The lesions occurred in 7 cases of the right pelvis, 6 cases of the left pelvis, and 2 cases of the bilateral pelvis. There were 38 lesions involving 3 or more parts in 7 cases, and 13 lesions invaded the surrounding soft tissue.All the 4 patients underwent X-ray examination showed multiple bone destruction, including 2 cases of single cystic expansive bone destruction, 1 case of irregular bone destruction with small bone shadow on the edge, and 1 case of cystic expansive bone destruction with patchy bone destruction; 4 cases of marginal osteosclerosis without periosteal reaction .Echinococcus granulosus is the main pathogen leading to hydatid disease in the pastoral areas of China, which seriously endangers human health and the development of animal husbandry. Granulosus has a carnivorous animal as a definitive host and an intermediate host such as sheep, cattle, and other herbivores. Humans are incidental hosts due to ingestion of the parasitic eggs from contaminated food or water. The larvae released from the eggs penetrate the intestinal wall and migrate to various organs. In addition to having a clear history of pastoral contact, a relatively higher prevalence has been noted in the rural populations that is mainly attributable to the slaughtering of animals in these areas [se areas . In thisse areas . Serologse areas . Routinese areas . In thisX-ray plain film is one of the preferred methods for the examination of pelvic cystic echinococcosis, which is characterized by continuous cystic bone destruction, irregular low-density shadow when invading soft tissue, and sometimes calcification which can be seen on the wall or inside the cyst. The involved sacroiliac joint or hip joint may narrow or disappear. The radiological appearance of pelvic hydatidosis varies and is influenced mainly by the location of the cyst, age of the cyst, and associated complications, such as secondary infection and rupture, so definite diagnosis sometimes needs to be combined with clinical history and further imaging examination. Anterior-posterior pelvic radiograph showed that most of the 4 cases had multiple cystic expansive bone destruction and all of them had marginal osteosclerosis and no periosteal reaction. The surrounding soft tissue was invaded, and the soft tissue around the involved bone showed low-density spherical shadow of different size, with or without calcification. The sacroiliac joint involvement showed hyperostosis at the lower edge of the sacroiliac joint and blurred joint surface. Among them, 2 cases had a history of hepatic echinococcosis and a history of pulmonary echinococcosis. It is basically consistent with the relevant research results , which sCT examination of pelvic cystic echinococcosis can not only precisely assess the type of bone destruction, calcification, and pathological fracture of echinococcosis , but alsIn addition, the recurrence rate of pelvic cystic echinococcosis is high, and the repeated course of disease can easily lead to secondary infection, or the interruption of treatment can lead to long-term wound healing, resulting in subcutaneous abscess or fistula , which rA total of 5 patients in this group underwent MRI examinations. As MRI examinations can observe the morphological characteristics, scope, location, and the relationship between the surrounding tissues and organs of bone cystic echinococcosis from the sagittal, coronal, and axial planes, it is of great value in the diagnosis of pelvic cystic echinococcosis. Single cystic echinococcosis of the pelvis showed intermediate or low signal intensity on T1-weighted images and hyperintensity on T2-weighted images in the involved bone and surrounding soft tissue on MRI  and highIn terms of management, hydatid disease of the pelvic bone is particularly a serious clinicopathologic entity as the cyst in this location may invade pelvic joints, which can potentially make complete recovery difficult. Although the definitive treatment of bony hydatidosis is surgery, a number of studies have highlighted the combination of antihelminthic chemotherapy and surgery as a feasible choice . In the Aneurysmal bone cyst, giant cell tumor, and bone tuberculosis can be considered as potential differential diagnoses in this case , 11. TheThe results of this study suggest that the lesions of pelvic cystic echinococcosis are mostly cystic expansive and osteolytic bone destruction, which is easy to invade the surrounding soft tissue, often accompanied with calcification. Among them, multiple cystic lesions are characteristic, which can improve the early diagnosis rate and achieve the purpose of early treatment.


Widespread use of antiretroviral treatment (ART) has led to decrease in the incidence of HIV/AIDS-related mortality. Besides the availability of ART, medication adherence is essential for treatment success. There is a scarcity of data reported from Turkey regarding ART adherence among people living with HIV/AIDS (PLWHA). Therefore, this study was undertaken to determine medication adherence and related factors among PLWHA in Turkey. The sample consisted of 158 PLWHA, who were being followed up at Infectious Diseases Outpatient Clinic of Hacettepe University Hospital. Data were collected using an individual questionnaire and the Turkish version of the Morisky Medication Adherence Scale. The median patient age was 38 years, 80.4% were male, and 51.3% were married. The median duration of both HIV infection and ART was 3 years. Sixty-one percent used two drug regimens. Sixty-one percent were highly adherent to ART while 37.9% were moderately adherent. The absence/presence of social support resources, disease duration, ART duration, and being informed about the ART regimen were statistically associated with medication adherence. Our results suggest that medication adherence is excellent among Turkish PLWHA. Interventions, including effective social support, and continuous counseling about ART, might further boost the adherence of PLWHA. There are 36.9 million people living with HIV/AIDS (PLWHA) in the world. Among them, 34.3 million are adults, and 17.4 million are women (1). According to the Turkish Ministry of Health’s National HIV Registry, about 17,000 people are living with the HIV infection in Turkey. However, due to the long latency, lack of adequate target population testing, and inadequate registry systems, this number may not be robust (2). From 2001 to 2013, the annual number of new HIV infections declined by 38% globally, followed by a significant decline in AIDS-related deaths. However, trends in new infections have differed among regions and countries (3), and the number of new HIV diagnoses increased 450% after 2012 in Turkey (2).There is no cure for HIV infection. Nonetheless, antiretroviral treatment (ART) can be implemented to overcome the infection-associated adverse outcomes . ART is the cornerstone of management of PLWHA. Widespread use of ART led to a decrease in the incidence of HIV/AIDS-related mortality both in the developed and developing world (6). Furthermore, ART improves the quality of life and prolongs the life expectancy (7).Medication adherence is defined as ‘‘the act of conforming to the recommendations made by the provider with respect to timing, dosage, and frequency of medication taking’’ (8). Besides the availability of ART, medication adherence must be at least 90% to suppress HIV replication (9–11). Medication adherence is crucial in maintaining therapeutic drug levels, ensuring virologic suppression, reducing drug resistance (12), preventing immunologic decay from HIV, and, especially, reducing the risk of HIV transmission (13). Variation in medication adherence has been reported among PLWHA by various studies. Data from Georgia, Iran, Russia, and Greece suggested that the proportion of the adherent PLWHA was 79.0%, 63.9%, 82.0%, and 81.8%, respectively (14–17). A small and older study composed of 36 PLWHA from Turkey found the ART adherence rate as 38.9% (18). Risk factors associated with low medication adherence were being busy, forgetfulness , side effects of ART, economic problems (8), stigmatization , and emotional stress . In addition, lack of central insurance and therefore high cost is another reason for low adherence (25). In Turkey, it is stated in the 67th article of the Social Insurance and General Health Insurance Law that people with any communicable diseases can benefit from the General Health Insurance system without any delay. HIV infection is also classified in this list. Hence, ART and other medical costs are fully covered by the government (26). PLWHA are charged only for a small amount of the expenditure per prescription. It is called the “contribution margin” and is 10% of drug price (27).It is important to evaluate ART adherence among PLWHA. There is a scarcity of data reported from Turkey regarding ART adherence. This study was undertaken to determine the medication adherence and related factors among PLWHA in Turkey.This hospital-based descriptive study was conducted at the HIV/AIDS Outpatient Clinic of Hacettepe University Hospital in Ankara between 7 September 2015 and 27 April 2016. Hacettepe University Non-invasive Ethical Research Board Study reviewed and approved the study protocol (Ref. No.: 16969557-981–02.09.2016). The researchers worked in collaboration with the clinicians in the identification of HIV/AIDS patients, who were receiving ART. A written statement was also included in the introductory part of the questionnaires, explaining the study purpose and confidentiality of the information collected. All data were obtained from respondents in a private room. A total 310 PLWHA were being followed up at the HIV/AIDS outpatient clinic. The calculated sample size was 138, assuming α = 0.05 and 1-β = 0.95 in power analysis. During the study period, 186 PLWHA applied to the infectious diseases clinic for treatment and follow-up. Among them, 158 PLWHA who fulfilled the inclusion criteria and voluntarily agreed to participate in the research were included in our study. The inclusion criteria were to be on ART for at least 6 months, at least 18 years old, and to provide informed consent. Exclusion criteria included not providing informed consent and not being on ART. A total of 28 patients were excluded, as they did not meet the inclusion criteria. As a result, 85% (n = 158) of the PLWHA who applied to the infectious diseases clinic were included in the final analysis (Table 1). Our results of the 158 study participants are representative of 310 PLWHA treated in the HIV/AIDS outpatient clinic.Data were collected using an individual information questionnaire and the Turkish version of the Morisky Medication Adherence Scale (MMAS). The personal information questionnaire was developed by the researchers, using PubMed and Elsevier databases . This questionnaire included 7 questions about the participant’s demographic characteristics  and 22 questions on medication and disease-related factors  The Turkish version of the MMAS was used to determine adherence to ART among PLWHA. The reliability and validity studies of the Turkish version of the MMAS were previously reported by Vural et al. (30). This scale consists of 6 items, each being a 2-point type, implemented in evaluating ART compliance through a face-to-face interview. Higher scores indicate lower adherence to ART. According to this scale, zero score implies high adherence, 1–2 points denote moderate adherence, and 3–6 points means noncompliant. After informing the patient about the study, the data report form was distributed, and correspondents asked to fill the questionnaire in a private room located at the outpatient clinic, by paying attention to their privacy. rs) allowed identifying whether there was an association between age/duration of illness/duration of medication/number of medicines used/daily dose amount, and the MMAS score. The Mann–Whitney U and Kruskal–Wallis tests were adopted to determine whether there was a statistically significant difference in the MMAS score among the groups . The statistical significance level was set at P < 0.05.All analyses were performed using SPSS version 21.0  and Microsoft Excel 2013. Numbers, percentages, median, and the chi-square (χ2) test were used in the data analysis. The Shapiro–Wilk test was used to determine whether the data were normally distributed. Spearman’s rho coefficient . The median age was 38 , 80.4% (n = 127) were male, and 51.3% (n = 81) were married. The median duration of both HIV infection diagnosis and ART were 3 years. Sixty-one percent (n = 97) used two drug regimens. Table 1 summarizes the demographic values.Nearly all the respondents (98.7%) thought that the drugs are beneficial for them, but 26.6% (n = 42) reported that they missed at least one dose because of being busy. In addition, 28.5% (n = 45) thought that the number of the pills taken daily are too many, and 29.7% (n = 47) had ART-related side effects. Also, 8.2% (n = 13) had problems affording the drugs, and 17.1% (n = 27) did not have information about the importance of adherence to ART and how to use the drugs. Furthermore, 42.4% (n = 67) had stigma-related problems.Sixty-one percent (n = 97) were highly adherent to ART while 37.9% (n = 60) were moderately adherent. Adherence varied among the age groups. Adherent PLWHA was 64.7%, 62.0%, 60.5%, and 54.5% in the age groups 18–25, 26–45, 46–60, and over 60 years, respectively. There were no significant differences in adherence among the age groups . Medication adherence was not associated with sex, age, marital status, education level, and the number of prescribed drugs in the ART regimen (P > 0.05) (Table 1).Medication adherence differed in PLWHA who were previously informed about ART and those who were not. Adherence among the preinformed and uninformed patients was 65.6% and 40.7%, respectively (Table 2). Those who had information on the usage and importance of ART were statistically more adherent than those who did not have such information . The presence of ART-related side effects did not affect adherence. There was no difference in medication adherence between patients with and without side effects   (Table 2). Conversely, a significant association existed between the presence of social support and medication adherence. PLWHA who confirmed the presence of social support were more adherent than the PLWHA who did not have social support   (Table 2). rs = 0.275 and 0.254, P ≤ 0.001, respectively) (Table 3).The relationships of the MMAS score with age, duration after diagnosis, duration of ART, number of medications prescribed, and number of pills taken daily were examined. A low positive correlation between duration after diagnosis, duration of ART, and the MMAS score was demonstrated . The variance in adherence rates may result from differences in the research method, characteristics of the cohort, the administrative structure of the healthcare facilities, and the social security policies of the countries. It may also arise from the economic status of the patients, because people living in Europe, Asia, Africa, and America have different socioeconomic levels.In previous studies, ART adherence was associated with sex (23), age (36), education , economic status , substance use , disease duration (37), stigma , ART-related side effects , satisfaction with healthcare staff , number of pills taken daily , social support , mental health , using reminders , and having information prior to ART (38). In addition, being busy, forgetfulness , being away from home , lack of central insurance , and adverse taste of medicines (45) were the most common reasons for low adherence. After examining the reasons for low adherence, according to the countries, the most leading causes included being busy, being away from home, and forgetfulness, in the USA; side effects, substance abuse, and stigma in Europe; stigma, forgetfulness, and substance abuse in Asia; and lastly, economic status, side effects, satisfaction with healthcare staff, and stigma in Africa. Mental health was the common factor that was associated with ART adherence in all regions. In our study, no correlations were observed between ART adherence and age, sex, marital status, education level, economic status, stigma, ART-related side effects, satisfaction with healthcare staff, and the number of pills taken daily. The fact that our sample is small, that the study is single-centered, and that we work at the referral center for HIV/AIDS may have affected these results. In addition, the small number of people with low adherence-related risk factors, such as lack of health insurance, dissatisfaction with healthcare personnel, and economic problems, may have contributed to these results.In this study, people who were previously informed about ART adherence had higher adherence rates compared to those who were uninformed. Our results agree with those of Schneider et al. who demonstrated a statistically significant relationship between having information about ART and ART adherence (46). Similarly, Demirkıran et al. (47) found that informing patients about medication adherence improves adherence among PLWHA. Providing information to PLWHA on ART and adherence ensures taking medicine regularly, without missing any dose, despite feeling good/bad, having side effects, being bored of the drugs, or feeling no need to continue the medication; thereby maintaining the high ART adherence level. Furthermore, in a routine daily outpatient care, every single PLWHA applying to our clinic was informed about ART and the disease itself. Interestingly, some of the PLWHA under our care were still denying previous education on ART and the disease. This finding restresses the importance of continuous medical briefing at every visit. Another factor that was significantly associated with ART adherence was the absence/presence of social support resources (Table 2) (P = 0.019). Kalichman et al. (48) observed that patients who had lack of support resources were less adherent to ART. In another study, in which pharmacists talked about their experience while working with PLWHA, lack of support resources was the major obstacle for ART adherence (25). Our findings are consistent with those of the previous studies in the literature. The absence of social support increases the feelings of anger, offense, despair, and depression in the later stages of HIV/AIDS infection. Consequently, all of these factors affect patients’ adherence to ART adversely. No statistical association was found between ART adherence and having ART-related side effects, being satisfied/dissatisfied about the healthcare professionals, stigma, using reminders, and economic status (P > 0.05) (Table 2). In contrast to our findings, PLWHA who have ART drug-related side effects , are dissatisfied about health care professionals (50) and had stigma-related problems  were found to be less adherent in other studies. From this perspective, our findings were inconsistent with those in the literature. These discrepancies may be because most of our respondents were satisfied with the healthcare professionals in the HIV/AIDS Outpatient Clinic. Also, the prescribed ART was current, and therefore most of the PLWHA did not experience ART-related side effects. Furthermore, we did not use a comprehensive tool to evaluate the stigma. Instead, it was asked with only one question in the questionnaire form. These reasons may explain the lack of association between stigma, ART-related side effects, being satisfied/dissatisfied with the healthcare professionals, and ART adherence.In our study, weak positive correlations were observed between disease duration, ART duration, and ART adherence (P = 0.001) (Table 3). It was found that taking ART for a long time could increase the motivation for medication adherence. Our findings corroborated the literature . When the duration of ART is long, PLWHA become used to the drugs, require fewer reminders, and the drugs become a regular part of their lives. All these factors are thought to maintain a high medication adherence among PLWHA.This work is the first extensive study from Turkey, determining ART adherence and related factors. Our study was undertaken in a tertiary level referral center, with strict case and outcome definition criteria. The prospective study design minimized the missing data. Our sample size is statistically sufficient to draw a conclusion on the ART adherence among Turkish PLWHA. Conversely, the single-center design, lack of a reliable stigma scale, and lack of pharmacy refill dates are main limitations in our study.In conclusion, the absence/presence of social support resources, disease duration, ART duration, and being informed about the ART regimen were statistically associated with medication adherence. Therefore, PLWHA must be continuously informed about the ART daily dosing regimen, as an intervention to maintain a high ART adherence. Suggesting reminders, ensuring and searching social support resources, and determining the factors that negatively affect ART adherence represent the other solutions and propositions for increasing ART adherence within the framework of nursing. Finally, considering that newly diagnosed patients and those who are new to ART have relatively low drug compliance, informing PLWHA on the use of ART should be further intensified in newly diagnosed individuals and those who have just started ART.This study was supported by a grant from the Yıldırım Beyazıt University Scientific Research Department (Project No.: 3049).


Poultries including chickens, ducks, geese, and pigeons are widely used in the biological and medical research in many aspects. The genetic quality of experimental poultries directly affects the results of the research. In this study, following electrophoresis analysis and short tandem repeat (STR) scanning, we screened out the microsatellite loci for determining the genetic characteristics of Chinese experimental chickens, ducks, geese, and pigeons. The panels of loci selected in our research provide a good choice for genetic monitoring of the population genetic diversity of Chinese native experimental chickens, ducks, geese, and ducks. Laboratory animals are important experimental materials for science research. They play key roles in the investigation of pathogenesis, diagnosis of diseases, pharmaceutical research, and other fields . The genPoultry, including chicken, duck, goose, and pigeon, has become commonly used laboratory animals . They arHence, in this study, we screened out the microsatellite loci with uniform distribution, stable amplification, and rich polymorphism in experimental chickens, ducks, geese, and pigeons with different genetic backgrounds . We deveThree outbred groups and three haplotype groups of experimental chicken were used in this research: outbred group BWEL-SPF chickens ((SCXK (black) 2017-005)), 40 samples, 37 weeks old, 6 males and 34 females, which has been closed for 20 generations; outbred group BM chicken (from BWEL chicken lineage (SCXK (black) 2017-005)), 40 samples, 14 weeks old, 6 males and 34 females; outbred group Beijing oil chickens, 46 samples. MHC haplotype chickens were bred from the 13th generation of BWEL chicken, the haplotype was continuously selected based on the MHC core genes, and the half-sibling or sibling mating method was used to breed to the 8th generation [Two outbred groups and four haplotype groups of experimental duck (bred from Jinding (JD) duck lineage (SCXK (black) 2017-006)) were selected: outbred group 1, 40 samples, 37 weeks old, 6 males and 34 females; outbred group JD duck, 40 samples, 37 weeks old, 6 males and 34 females; 10 A haplotype ducks, 53 weeks old, 1 male and 4 females; 10 B haplotype ducks, 53 weeks old, 1 male and 4 females; 10 C haplotype ducks, 53 weeks old, 1 male and 4 females; 10 D haplotype ducks, 53 weeks old, 1 male and 4 females. All the samples are duck muscle tissue and were from HVRI, CAAS.We collected two outbred groups of experimental geese: outbred group Guangdong Wuzong goose, 44 samples, 37 weeks old, 6 males and 34 females; outbred group Yangzhou goose, 44 samples, 37 weeks old, 6 males and 34 females. All the samples are goose liver tissue. Guangdong Wuzong geese were from Southern Medical University, and Yangzhou geese were from Yangzhou University.Forty pigeons were randomly selected from two populations of white king pigeons and silver king pigeons, half male and half female, with no age limit. All the animals were from Liujinlong pigeon farms in Beijing. Their heart tissues were collected.All breeding is carried out in accordance with Chinese agricultural standards NY/T 1901. What is more, all experiments followed the 3R principle.By searching PubMed and using SSR Hunter software to analyze animal gene information, we obtained microsatellite loci for further screening.μL, stored in -20°C.Phenol-chloroform extraction method was used to extract DNA from muscle, liver, and heart tissue. TIANamp Blood DNA Kits  were used to extract DNA from chicken blood samples. All DNA concentrations were diluted to 50 ng/μL reaction volume containing 10 μL Dream Taq Green PCR Master Mix , 2 μL pure water (ddH2O), 10 pmol each primer, and 50 ng of the extracted DNA template. The PCR protocol was as follows: 94°C for 5 min, followed by 35 cycles of 94°C for 30 s, suitable temperature for 30 s, 72°C for 30 s, and a final extension at 72°C for 5 min. Amplified products were stored at -20°C for further analysis.The PCR was performed in a 20 Amplified products were electrophoresed on a 2% agarose gel at 130 V, 90 min.We performed STR scanning on PCR amplification products of candidate loci. The forward primers of candidate microsatellite loci were fluorescent labelled with FAM, HEX, and TAMRA. The sample genome was amplified with fluorescent primers, and the amplified products were scanned by STR through 3730xl DNA Analyzer . All the STR scanning was performed by Beijing Tianyi Huiyuan Biotechnology Co., Ltd.GeneMarker V2.2.0 software was used to analyze the length of amplified fragments from different populations at each microsatellite locus. Popgene 3.2 software was used to analyze the observed number of alleles, effective number of alleles, Shannon's information index, and effective heterozygosity of microsatellite loci. The polymorphic information content of multiple sites was calculated using PIC calculation software (PIC_CALC.0.6).Firstly, we obtained the microsatellite locus information of experimental chickens, ducks, geese, and pigeons by searching previous reports on PubMed and using the SSR Hunter software to analyze the genetic information of different populations , 15. We In order to clarify the amplification conditions of the microsatellite loci and exclude the loci with poor specificity, we performed temperature gradient PCR and agarose gel electrophoresis of microsatellite loci. Then, we performed PCR amplification on the most suitable conditions and subjected the PCR products to agarose gel electrophoresis to screen out loci with suitable length, good polymorphism in outbred groups, good monomorphism in haplotypes, and high specificity. Taking the chicken GGNCAMZO locus and duck AY264 locus as example, the results are shown in In summary, we selected 37 and 32 microsatellite loci with good polymorphism in the outbred groups and haplotypes of chicken, respectively , 16, 17.μT47 locus as an example, it showed polymorphism in the outbred group of pigeon . Taking the UU-Clif pigeon .We finally determined that in experimental chickens, 28 loci were selected for genetic monitoring in the outbred groups and 14 loci for haplotypes. All microsatellite DNA loci are shown in In experimental duck populations, we chose 25 loci and 15 loci for genetic monitoring in the outbred duck groups and haplotype groups. There are 12 common loci. Microsatellite loci are shown in 14 microsatellite loci with good polymorphism were considered as microsatellite markers in the outbred group of goose. In the outbred group of pigeon, we finally screened out 16 microsatellite loci with good polymorphism, several alleles, and typical stutter peaks. All microsatellite locus information is shown in We inputted the results of STR scanning into Popgene 3.2 to analyze experimental chicken in the outbred groups and the haplotypes at 29 loci. In the outbred groups, 28 microsatellite loci show a high degree of polymorphism, and the average number of observed alleles is 4.571. The average number of effective alleles is 3.270, and the average Shannon's information index is 1.198 . FurtherIn the other 3 haplotype populations, 14 microsatellite loci showed monomorphism in each population but showed different lengths in different haplotype populations. The average number of observed alleles is 1.571. The average number of effective alleles, the average Shannon's information index, and the average effective heterozygosity are 1.433, 0.316, and 0.207, respectively . The speIn the outbred group of duck, 25 microsatellite loci show polymorphism. The average number of observed alleles is 7.520, and the average number of effective alleles in the population is 4.162. The average Shannon's information index is 1.574, and the average effective heterozygosity is 0.683. The average PIC is 0.698. These data showed that in the outbred groups, the genetic diversity of microsatellite DNA loci is better, and the genetic diversity of each locus is quite different. The specific results are shown in In 4 haplotype populations, 15 microsatellite loci show monomorphism in each population. The average number of observed alleles is 4.133, the average number of effective alleles is 2.863, and the average Shannon's information index is 1.153, indicating that the genetic diversity of the loci in these haplotype populations is poor; the average effective heterozygosity is 0.500, indicating that the heterozygosity difference is small and the genetic information of the selected loci is relatively single. See In the outbred colony of experimental goose, 14 loci were selected. The average number of observed alleles, the average number of effective alleles, the average Shannon's information index, the average effective heterozygosity, and the PIC are 4.714, 3.038, 1.195, 0.528, and 0.582, respectively. The microsatellite loci have large interindividual differences within the population, and the population has high gene stability .The selected microsatellite loci all show good polymorphism in the experimental outbred pigeon populations. A total of 16 loci were selected. The average number of observed alleles is 7.875. The average effective allele number is 4.554; the average Shannon's information index and the average effective heterozygosity are 1.559 and 0.649. The average PIC is 0.674 .Among the three outbred chicken groups, the mean number of observed alleles, the mean number of effective alleles, the mean Shannon's information index, and the mean effective heterozygosity are shown in In the haplotype chicken populations, the highest mean observed number of alleles is observed in G7groups. Haplotype G7 has the highest mean effective allele number and the highest mean Shannon's information index. The mean effective heterozygosity of haplotype G7 is 0.364. The genetic heterozygosity of the 3 populations is very low, and the consistency is good .In the two outbred groups of duck, the mean number of observed alleles, the mean effective number of alleles, the mean Shannon's index, and the mean effective heterozygosity of outbred group 1 are higher than those of outbred group JD, indicating that outbred group 1 had better diversity. The results are shown in In the two outbred groups of goose, the mean number of observed alleles, the mean effective number of alleles, and the mean Shannon's index of Guangdong Wuzong goose are higher than those of Yangzhou goose, indicating that Guangdong Wuzong goose has a better diversity .The analysis of the two main experimental pigeon populations used for scientific research shows that the mean effective heterozygosity of two populations is 0.647 and 0.651, respectively. The mean number of observed alleles, the mean effective number of alleles, and the mean Shannon's index are higher in white king pigeons than in silver king pigeons. The comparison of the data is shown in Poultries are widely used and are indispensable supporting conditions for the life sciences and biomedicine industries. Specific pathogen-free (SPF) chicken embryos are used in the manufacture and quality control of biological product ; ducks pCoat colour gene testing method, biochemical marker gene testing method, immune marker gene testing method, and DNA molecular marker method are popular methods for genetic monitoring. Microsatellite DNA, mitochondrial DNA (mtDNA), restriction fragment length polymorphism (PCR-RFLP), single-stranded conformation polymorphism (PCR-SSCP), and specific gene polymorphisms are commonly used DNA molecular marker methods –27. AmonMicrosatellite DNA is characterized by short tandem repeats (STRs) of 1 to 6 nucleotides in eukaryotic genome with a random manner . It has The mean effective number of alleles is an indicator of genetic variation and mutation drift balance. In our study, Beijing oil chicken has the highest mean effective allele number of three outbred chicken populations; outbred duck group 1 has higher mean effective allele number than outbred duck group JD. The outbred goose group Guangdong Wuzong and outbred pigeon group white king have the highest mean number of effective alleles in outbred goose populations and outbred pigeon populations, respectively. The higher mean effective number of alleles indicates that the population can maintain the original gene and avoid new variations under the pressures from genetic drift and artificial selection. The results show that Beijing oil chicken, outbred duck group 1, Guangdong Wuzong goose, and white king pigeon are the most stable strains in the outbred group of experiment chicken, duck, goose, and pigeon groups in this research, respectively.The mean effective heterozygosity of a population is an important indicator of population genetic diversity and can reflect the richness of the detected genes. It is generally believed that when the mean effective heterozygosity of the population is less than 0.5, it indicates that the individual differences in the population are small and the genetic heterozygosity is low, which does not conform to the genetic characteristics of an outbred group animal. When the mean effective heterozygosity of the population is higher than 0.7, its genetic diversity is high .Hence, we found that the mean effective heterozygosity of BWEL, BM, and Beijing oil chicken groups is all greater than 0.5, which conforms to the characteristics of the outbred group. The mean effective heterozygosity of BWEL and BM chicken groups is nearly 0.5. The average effective heterozygosity of G1, G2, and G7 groups is all less than 0.5. It is also consistent with the background that BWEL, BM, and Beijing oil chickens are outbred colonies; Beijing oil chicken has abundant genetic diversity and high selection potential for it has the highest mean effective heterozygosity among the outbred chicken groups in this study. This may be due to the large population. Duck group 1 and JD duck all have a mean effective heterozygosity greater than 0.680 which indicates a high genetic diversity. The mean effective heterozygosity of Guangdong Wuzong goose group, silver king pigeon group, and white king pigeon group is all greater than 0.5 which reflects abundant genetic diversity. The mean effective heterozygosity of three haplotype chicken groups and four haplotype duck groups is 0.207 and 0.500, respectively. The result indicates a good consistency in haplotype chickens and ducks. This may be the result of long-term full-sib and half-sib reproduction. Chickens and ducks are more widely used in biological research, and the breeding standards are stricter, while geese and pigeons are more useful in agriculture. Haplotype chickens have lower mean effective heterozygosity than haplotype duck populations, which is consistent with a longer history of breeding in experimental chickens.When measuring the degree of gene variation, PIC is often used as a variation index. It is generally believed that when PIC is between 0.25 and 0.5, it is moderately polymorphic, and <0.25 shows a low level of polymorphism, when PIC is greater than 0.5, it means a high level of polymorphism . In our In conclusion, we identified appropriate microsatellite marker systems for native experimental chickens, ducks, geese, and pigeons in China. The combination of loci selected in our research provides a good choice for genetic monitoring of the quality and the population genetic diversity of poultry stocks.


At the beginning of June 2020, there were nearly 7 million reported cases of coronavirus disease 2019 (COVID-19) worldwide and over 400,000 deaths in people with COVID-19. The objective of this study was to determine associations between comorbidities listed in the Charlson comorbidity index and mortality among patients in the United States with COVID-19.n = 17,155) were female. The most common comorbidities listed in the Charlson comorbidity index were chronic pulmonary disease  and diabetes mellitus . Multivariate logistic regression analyses showed older age , male sex , being black or African American compared to white , myocardial infarction , congestive heart failure , dementia , chronic pulmonary disease , mild liver disease , moderate/severe liver disease , renal disease , and metastatic solid tumor  were associated with higher odds of mortality with COVID-19. Older age, male sex, and being black or African American (compared to being white) remained significantly associated with higher odds of death in age-stratified analyses. There were differences in which comorbidities were significantly associated with mortality between age groups. Limitations include that the data were collected from the healthcare organization electronic medical record databases and some comorbidities may be underreported and ethnicity was unknown for 24% of participants. Deaths during an inpatient or outpatient visit at the participating healthcare organizations were recorded; however, deaths occurring outside of the hospital setting are not well captured.A retrospective cohort study of adults with COVID-19 from 24 healthcare organizations in the US was conducted. The study included adults aged 18–90 years with COVID-19 coded in their electronic medical records between January 20, 2020, and May 26, 2020. Results were also stratified by age groups . A total of 31,461 patients were included. Median age was 50 years  and 54.5%  has led to a public health emergency internationally.As of June 2020, there were over 400,000 deaths reported with COVID-19 globally and over 110,000 deaths were in the US, but many people have also recovered.Because of the unprecedented outbreak of COVID-19 worldwide, little is known about which underlying health conditions may impact a person’s likelihood of dying with COVID-19.Some previous studies have suggested being older; being from a black, Asian, or minority ethnic (BAME) background; and having certain health conditions may increase risk of death with COVID-19, but further evidence is needed to understand factors which influence this.The research utilized a network of 24 healthcare organizations in the US, which provided deidentified data from electronic medical records of patients.A total of 31,461 adults with COVID-19 coded in their electronic medical records were included in the study after a search of the network between January 20, 2020, and May 26, 2020.We determined associations between age, sex, ethnicity, comorbidities, and death with COVID-19 during the study period.After accounting for the other included factors in the study, being older, being male, being black or African American, and having a history of myocardial infarction, congestive heart failure, dementia, chronic pulmonary disease, mild liver disease, moderate/severe liver disease, renal disease, or metastatic solid tumor were all associated with higher odds of death with COVID-19.There were differences in which comorbidities were associated with death when we stratified the results by age group.Identifying factors associated with death with COVID-19 could help with hypothesis generating for clinical trials and identify patients who may need to be targeted for early intervention or monitoring.The study has limitations; for example, some health conditions may be underreported or incorrectly coded in electronic medical records at the time of data entry, and all deaths of participants may not have been captured. Coronavirus disease 2019 (COVID-19) was first reported in Wuhan, China, in December 2019 .An emerging evidence base has started to identify factors that associate with adverse outcomes for people with COVID-19. Older age is the most consistent risk factor for severity of COVID-19 that has emerged from the literature so far –6. ThereMore studies are needed to determine associations between comorbidities and outcomes for patients with COVID-19. Multimorbidity is closely linked to frailty status , which iThe study used data from TriNetX, a global federated health research network that provided an anonymized dataset of electronic medical records (EMRs). The TriNetX network was searched on June 9, 2020, and a deidentified dataset of patients with COVID-19 aged up to 90 years identified in EMRs between January 20, 2020, and May 26, 2020, was provided. The data on the research network come from academic medical centers, specialty physician practices, and community hospitals. Further details about TriNetX processes and standardization of data are in Patients with COVID-19 were identified following criteria provided by TriNetX based on Centers for Disease Control and Prevention (CDC) coding guidelines . PatientThe study included all patients with COVID-19 recorded in their EMRs from participating healthcare organizations. This included both inpatient and outpatient care settings, but the type of visit was not well recorded. History of comorbidities listed in the Charlson comorbidity index were identified if the patient had a corresponding ICD code for the condition since January 1, 2015, in their EMRs captured in TriNetX . The timp < 0.05 were planned to be inserted into a forward multivariate logistic model. All variables were statistically significant in unadjusted analysis and inserted to the multivariate model, apart from in age-stratified analyses. No imputations were made for missing data. Data were requested from TriNetX and all analyses were conducted with Stata v.14.0.Descriptive statistics included proportions for categorical variables and medians and interquartile ranges (IQRs) for continuous variables. Unadjusted and multivariate logistic regressions were performed to explore associations between age, sex, ethnicity, comorbidities, and mortality. Any deaths during the study period captured in EMRs of the participating healthcare organizations were included in the analysis. Variables identified as statistically significant predictors with a significance level of This study is reported as per the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) guidelines  18]. No. No18]. No ethical approval was obtained, as no patient identifiable information was received and the data were analyzed anonymously.n = 9,040) in the Northeast, 21%  in the Midwest, 30%  in the South, 21%  in the West, and 0.5% (n = 177) unknown. Of the patients identified, 50.3%  were identified from ICD codes only ; 15.2%  were identified from the ICD codes B97.29 or B34.2 only.In total 33,488 patients from 24 healthcare organizations had 1 or more of the specified COVID-19 codes or a positive laboratory test in their EMRs during the study period. Patients in the cohort were distributed between the 4 large Census Bureau–designated regions of the US as follows: 27% (n = 670) did not have age or sex recorded within the TriNetX network and were excluded from analyses. A further 1,357 patients were aged <18 years and were also excluded. Therefore, 31,461 patients were included in analyses. The median (IQR) age was 50 (35–63) years and 54.5%  were female. In total, 45.2%  of participants were white, according to their EMRs, and 27.8%  were black or African American; smaller proportions of people from other ethnicities were identified, and ethnicity was unknown for 23.8% . Of the total cohort, 59.5%  of patients did not have a record of any of the comorbidities listed in the Charlson comorbidity index; 17.4%  had 1 comorbidity listed, 7.9%  had 2 comorbidities, and 15.2%  had ≥3 comorbidities. The most common comorbidities were chronic pulmonary disease  and diabetes mellitus   .n = 1,669) had no end date recorded on a healthcare organization encounter during the study period and therefore may have not yet been discharged. The median (IQR) estimated time in the study was 54 days (36–68). During the study period, 4.1%  patients were recorded as deceased, and the estimated median (IQR) time to mortality after first recording of COVID-19 was 9 days (4–17). Of the total patient cohort, 4.1%  had a procedure code indicating invasive mechanical ventilation during the study period in EMRs. Of the patients who died, 49.5% (n = 642) had a procedure code for invasive mechanical ventilation compared to 2.2% (n = 654) of patients who were not recorded as deceased.Of the 31,461 patients, 5.3% (p < 0.001), male sex , being black or African American compared to white , myocardial infarction , congestive heart failure , dementia , chronic pulmonary disease , mild liver disease , moderate/severe liver disease , renal disease , and metastatic solid tumor . Being native Hawaiian or other Pacific Islander was associated with higher odds of death compared to being white, but there were only 115 people identified as native Hawaiian or other Pacific Islander in this cohort. Having an unknown ethnicity was associated with reduced odds of death compared to being white .In unadjusted analyses, the following variables were associated with higher odds of death: older age, male sex, being black or African American compared to white, and all comorbidities investigated . Being ADifferences between age groups were found when results were stratified by age group . For all age groups, older age, being male, and being black or African American compared to being white were associated with higher odds of death in multivariate analyses. When examining comorbidities, history of myocardial infarction and renal disease were associated with higher odds of death for all age groups, but there were differences across age groups for other comorbidities and associations with mortality. For people aged <50 years, history of mild liver disease and any malignancy were associated with higher odds of death. For people aged 50–69 years, history of congestive heart failure, chronic pulmonary disease, moderate/severe liver disease, metastatic solid tumor, and AIDS/HIV were all associated with higher odds of death. For people aged 70–90 years, history of congestive heart failure and dementia were associated with higher odds of death .In this retrospective cohort study, we identified 31,461 adults with confirmed COVID-19 from EMRs from 24 healthcare organizations in the US between January 20, 2020, and May 26, 2020. Older age, male sex, black or African American ethnicity, and a history of myocardial infarction, congestive heart failure, dementia, chronic pulmonary disease, mild liver disease, moderate/severe liver disease, renal disease, and metastatic solid tumor were associated with higher odds of mortality after adjustment for these factors and other comorbidities.Older age has been frequently reported to be an important factor associated with disease severity or mortality in patients with COVID-19 –6. ThereAge-stratified analyses in the current study highlighted the potential importance of considering age groups when examining associations between comorbidities and mortality with COVID-19. We found a history of myocardial infarction or renal disease to be associated with mortality in all age groups, but there were differences found between age groups for which other comorbidities associated with mortality. In this study, a borderline association was found in this study between AIDS/HIV and mortality with COVID-19 when examining all adults, but age-stratified analyses suggested AIDS/HIV was associated with significantly higher odds of mortality only in people aged 50–69 years. Patients with AIDS/HIV represented a small proportion of the total patient population, and larger studies of patients with AIDS/HIV and COVID-19 are needed to further determine the association. The findings in this study of an association between liver disease and mortality for patients with COVID-19 are in line with a recently published study that used data from TriNetX and showed patients with preexisting liver disease had a higher risk of mortality with COVID-19 .This study has several limitations. The data were collected from healthcare organization EMR databases and some comorbidities may be underreported, and ethnicity was not available for all participants. Residual confounding may include lifestyle factors and socioeconomic status, which were not available from EMRs. Body mass index was only available for approximately 5% of participants and so was not included in analyses. We could also not determine the influence of attending different healthcare organizations, owing to data privacy restrictions. In these analyses, we found a significant association between those with “unknown” ethnicity from EMRs and reduced odds of death compared to white patients. Participants with unknown ethnicity may have been participants who did not fit in the limited prespecified ethnicity categories within TriNetX, but this could not be explored further, given restrictions on data privacy. Only age at death, not date of death, was available in downloaded data, so time-to-event analyses could not be performed. Participants who died with COVID-19 after the study end date would have been recorded as alive in the present analyses, and deaths outside of the participating healthcare organizations are not well captured. The data in EMRs are susceptible to errors in coding or data entry when patient information is translated to ICD codes. In this study, we looked at any history of the comorbidities identified in the Charlson comorbidity index since 2015. We could not determine if the patient was no longer living with the condition. Recording of ICD codes in administrative data may vary by factors such as age, number of comorbidities, severity of illness, length of hospitalization, and whether in-hospital death occurred . The datIncreasing age, being male, being black or African American compared to white, and a history of myocardial infarction, congestive heart failure, dementia, chronic pulmonary disease, liver disease, renal disease, and metastatic solid tumor were associated with mortality in adults with COVID-19. There were differences in which comorbidities were associated with higher odds of mortality depending on age group.S1 STROBE ChecklistSTROBE, Strengthening the Reporting of Observational Studies in Epidemiology.(DOCX)Click here for additional data file.S1 TableCOVID-19, coronavirus disease 2019.(DOCX)Click here for additional data file.S2 TableCOVID-19, coronavirus disease 2019.(DOCX)Click here for additional data file.S3 TableCOVID-19, coronavirus disease 2019.(DOCX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.S1 Data(XLS)Click here for additional data file.


Nuclear receptors (NRs) are highly relevant drug targets in major indications such as oncologic, metabolic, reproductive, and immunologic diseases. However, currently, marketed drugs designed towards the orthosteric binding site of NRs often suffer from resistance mechanisms and poor selectivity. The identification of two superficial allosteric sites, activation function-2 (AF-2) and binding function-3 (BF-3), as novel drug targets sparked the development of inhibitors, while selectivity concerns due to a high conservation degree remained. To determine important pharmacophores and hydration sites among AF-2 and BF-3 of eight hormonal NRs, we systematically analyzed over 10  Nuclear receptors (NRs) are ligand-inducible transcription factors that are attractive drug targets due to their involvement in a multitude of physiological and pathological processes. Currently marketed drugs designed to interact with the buried ligand binding pocket (LBP) of the respective receptor are used in major indications such as oncologic, metabolic, reproductive, and immunologic diseases ,2. HowevWhile several structures with cocrystallized ligands have been determined for the AR, targeting superficial binding sites such as AF-2 and BF-3 of other receptors remains a challenge due to their comparably large size, shallowness, and high flexibility ,31. Knowruggable ,38. Anotruggable ,40,41,42ruggable .Here, we applied cosolvent MD simulations, hydration site prediction, and molecular docking to a set of eight NRs including AR, ERIn contrast to previous works, our objective was the systematic determination of the main pharmacophores and positions of structural waters in order to compare them within our selection of human hormonal NRs to ultimately navigate the design of potent and selective inhibitors for each particular receptor.The structures of LBD of AR, ERved fold B. We conved fold . While tTo this date, the AR is the most intensively studied NR regarding the development of allosteric inhibitors, due to its involvement in the genesis and progression of prostate cancer, which is one of the leading causes of cancer-related death in men. ,13. Evenreceptor . Unfortureceptor , which lreceptor ,37,38.The aforementioned work inspired us to systematically apply this simulation protocol to eight NRs which are known to suffer from poor drug selectivity ,24,27. Bα A. In accine core . In consine core . Notablyhibition . Therefodication . While tdication , our resAs mentioned before, the BF-3 site of the studied NRs displayed a higher degree of heterogeneity regarding the cosolvent densities A. While the site , which i site . T. Tβ AF-2eceptors . Analogoeceptors , are proThe design of numerous allosteric inhibitors considered in this study was itself assisted by computational chemistry methods such as virtual screening ,13,17,24observed D,E. Whil2 series . The sco2 series  confirmeEven though the majority of studies neglected off-target binding of their compounds, one study extensively evaluated their inhibitors against other NRs overlapping with our selection . Althougeraction  between ction [α . The chaAfter a sequence alignment in the UGENE v1.32.0  suite usAll ligands were retrieved from various publications that evaluated their compounds and provided evidence for binding to either of the allosteric sites . Compounep panel  within tep panel  using thep panel  at physiThe protein structures used in this study were retrieved from the Protein Data Bank  and prepdatabase  and, in The simulations in pure water were conducted using the Desmond simulation engine (v.2019-3) . Using tFor the cosolvent MD simulations, we used the Mixed Solvent MD workflow that comes with the Desmond simulation engine . As probworkflow  was condTo quantify the conformational change of the AF-2 and BF-3 sites induced by the probe molecules, we determined the heavy-atom RMSD between representative structures of the cosolvent simulations and the pure water simulations of the individual residues located in the allosteric sites. First, we determined the representative structure of the cosolvent simulations by inputting the last frame of each simulation for each probe into the MaxCluster algorithm and selecting the structure with the highest rank according to the 3D-jury score . SimilarThe simulations to determine the hydration sites were performed using the WATsite 3.0 protocol ,42 that n set to 2.0, similar to other protocols [For each receptor, all crystal structures with a resolution below 2.5 Å were retrieved from the Protein Data Bank and superimposed. Next, only the water molecules were kept in the structures and merged into a single PDB file that was used as input for an in-house python routine that determined the cluster centroids along with their occupancy using the DBSCAN algorithm with an epsilon value of 0.9 and rotocols . ClusterWe used the Glide protocol ,58 to dohttps://github.com/mmodbasel/NR_allosteric_sites. Structure files with the determined hydration sites using both WATsite and the crystal structure analysis are available in PDB format together with files that allow the cosolvent densities to be viewed in CNS format, which is compatible with PyMol.All described in-house python routines and files are available in our public repository at Several allosteric inhibitors for the AR proved their efficacy in blocking AR signaling through experiments with cells or xenograft in vivo studies ,8,17. Be