Moreover, patients who ingested foods rich in SFA, according to FFQ-6, had a higher mortality risk . The hazard ratio for mortality associated with increased SFA content equalled 4.47 (1.63–12.26). Addition of age and inflammation (hsCRP > 5 mg/L) into the Cox model did not modify this relationship. However, SFA content turned out to be significantly higher in patients with diabetes mellitus and cardiovascular disease, as compared to patients free from these co-morbidities. Their addition to the model attenuated the relationship between SFA and mortality, making it statistically insignificant. Conclusion: The serum content of SFA turned out to be a strong predictor of mortality in dialysis patients. However, given the significant associations between SFA, DM, and CVD, interventional studies with controlled SFA intake are needed to evaluate the causal links between SFA, co-morbidities and survival.Background: Cardiovascular mortality in dialysis population remains very high. Saturated fatty acids (SFA) contribute to atherosclerosis and to cardiovascular risk. Aim: The aim of this study was to evaluate the relationship between mortality in dialysis patients and the serum SFA content. Methods: Survival of 54 patients on dialysis was assessed. A total of 21 SFA from patients’ sera were measured by gas chromatography-mass spectrometry (GC-MS). Diet was assessed by food frequency questionnaire FFQ-6. The SFA content is presented as fatty acid proportion (%). Results: During the observation time (median 66 months) 22 patients died. There was a significant relationship between elevated SFA (above SFA mean) and mortality (log-rank 3.13;  The prevalence of chronic kidney disease (CKD) is estimated at a rate as high as 6–13% of the general population . MoreoveIn the general population, lipid disorders are among the most acknowledged risk factors for CVD. In dialysis patients, atherosclerotic dyslipidaemia is prevalent. Hypertriglyceridemia and low-HDL cholesterol concentration are the most typical and potentially deleterious lipid abnormalities in the course of CKD . HoweverFatty acids, carboxylic acids with an aliphatic chain, either saturated or unsaturated, play numerous crucial functions in the organism mainly as energy sources and membrane constituents. Saturated fatty acids (SFA) have only single bonds in their chains. They are mainly obtained through dietary intake of animal fats. In the general population, SFA have been linked to increased cardiovascular risk. Leading to an increase in low density lipoprotein (LDL) cholesterol concentration, they are thought to promote atherosclerosis, and hence, cardiovascular disease and mortality risk ,7. HowevThe aim of this study was to evaluate the relationship between mortality in dialysis patients and the serum content of saturated fatty acids (SFA).n = 27) was undergoing hemodialysis (HD) treatment, while the other half was on peritoneal dialysis (PD). Presence of diabetes mellitus (DM) and CVD was determined on the basis of patients’ medical charts.The study was performed based on a group of 54 ESRD patients, treated with dialysis in a single university-based dialysis centre. Exclusion criteria were as follows: intake of fatty acids supplements, acute inflammatory state, and unwillingness to participate in the study. Half of the group (v/v) following the method by Folch et al. [Blood was collected prior to an HD session in HD patients and during a routine check-up visit in PD subjects. Serum was stored at −80 °C, until analysed. A total of 21 SFA from patients’ sera were measured by gas chromatography-mass spectrometry (GC-MS). Total lipids were extracted from whole serum samples in a chloroform–methanol mixture , the most common dietary assessment tool used in large epidemiologic studies of diet and health and validated for the national population .Protocol of the study received approval from the Local Bioethics Committee at the Medical University of Gdansk (protocol no. NKEBN/614/2013-2014) and informed consents were obtained from all the patients.p value < 0.05 was considered to be statistically significant. Comparisons between two groups were assessed with a Student’s t-test, or a Mann–Whitney test, as appropriate. To assess correlations among the evaluated variables, Pearson’s correlation coefficient (r) was used. Survival analyses were made with the Kaplan–Meier survival curve and the Cox proportional hazard model, presenting data as hazard ratio ). Statistical processing of the results was performed with the use of the statistical software Statistica PL version 13.0 .Results are expressed as mean and standard deviation or median and interquartile range, as appropriate. The assumption of normality was verified with the Kolmogorov–Smirnov test. A p = 0.0017) (p = 0.03) (p = 0.03), and was elevated in patients with CVD when compared to patients without it . In Cox regression, the addition of age and inflammation (hsCRP > 5 mg/L) into the model did not modify the relationship between high SFA and mortality. However, addition of DM and CVD attenuated this relationship, making it statistically insignificant; HR 2.47 (0.78–7.71). According to food questionnaires, SFA content was associated with ingestion of food rich in saturated fat, although these relationships did not reach statistical significance. No associations between SFA and lipoprotein subclasses or with total cholesterol were observed. Sub-group analyses in patients stratified according to the use of HMG-CoA inhibitors revealed comparable results . During the observation, 22 patients died. Kaplan–Meier analysis revealed a significant relationship between elevated SFA (above SFA mean) and mortality  . In the  = 0.03) . However = 0.03) . Indeed, results . There w results .In the present study, the serum content of SFA in dialysis patients turned out to be a strong predictor of mortality. However, since it was heavily dependent on the presence of DM and CVD, including these co-morbidities into the analysis attenuated the associations between SFA and outcome to a statistically insignificant level. Moreover, patients who ingested foods rich in SFA, according to FFQ-6, had a higher mortality risk in our analysis. Similarly, this association lost its significance after accounting for the above confounders.In the general population, SFA intake and their content is generally believed to promote atherosclerosis, and in consequence, the cardiovascular risk . The majn = 13,614 participants) estimated a CVD risk reduction of about 10% for each 5% energy replacement; an effect comparable to that predicted from the effects of the intervention on TC:HDL cholesterol ratio [Cardiovascular disease is responsible for the largest proportion of deaths worldwide, and fatty acid profile is a potentially modifiable risk factor for the development of CVD, e.g., through dietary modifications; hence studies evaluating this issue seem of utmost clinical importance. Most public health dietary guidelines recommend limiting SFA intake ,16,17. Tol ratio . This maol ratio .n = 347,747 participants) did not demonstrate significant associations between the intake of SFA and the risk of chronic heart disease and/or stroke [n = 344,696 persons) demonstrated that replacement of SFA with polyunsaturated fat, but not carbohydrate or monounsaturated fat, was associated with decreased CVD risk [However, there are also opinions neglecting the role of SFA in CVD risk. A meta-analysis summarizing data related to SFA and CVD from 21 prospective epidemiologic studies , as well as patients treated with lipid-lowering therapy. Patients treated with PD are characterized by different CV risk factors than HD patients. For instance, they have a tendency for higher cholesterol concentrations because of the dialysis-associated glucose load, while HD patients are subject to lower hemoglobin concentrations because of blood losses during the dialysis procedure. Still, the CV mortality and the overall outcome are comparable with these two dialysis methods, as are the baseline characteristics of patients in our study who were stratified according to their treatment modality . SimilarIn conclusion, in our cohort of patients undergoing dialysis a strong relationship between serum SFA content, as well as SFA intake and survival, has been demonstrated. This association turned out to be significantly dependent on the co-existing illnesses that themselves constitute strong risk factors for poor outcomes. Whether SFA contributes to these diseases or acts just as innocent by-standers, can be determined solely through interventional studies.


Molecular animations play an increasing role in scientific visualisation and science communication. They engage viewers through non-fictional, documentary type storytelling and aim at advancing the audience. Every scene of a molecular animation is to be designed to secure clarity. To achieve this, knowledge on design principles from various design fields is essential. The relevant principles help to draw attention, guide the eye, establish relationships, convey dynamics and/or trigger a reaction. The tools of general graphic design are used to compose a signature frame, those of cinematic storytelling and user interface design to choreograph the relative movement of characters and cameras. Clarity in a scientific visualisation is reached by simplification and abstraction where the choice of the adequate representation is of great importance. A large set of illustration styles is available to chose the appropriate detail level but they are constrained by the availability of experimental data. For a high-quality molecular animation, data from different sources can be integrated, even filling the structural gaps to show a complete picture of the native biological situation. For maintaining scientific authenticity it is good practice to mark use of artistic licence which ensures transparency and accountability. The design of motion requires knowledge from molecule kinetics and kinematics. With biological macromolecules, four types of motion are most relevant: thermal motion, small and large conformational changes and Brownian motion. The principles of dynamic realism should be respected as well as the circumstances given in the crowded cellular environment. Ultimately, consistent complexity is proposed as overarching principle for the production of molecular animations and should be achieved between communication objective and abstraction/simplification, audience expertise and scientific complexity, experiment and representation, characters and environment as well as structure and motion representation. Modern technology makes video an easily accessible and therefore omnipresent medium in our lives and therefore also in the fields of knowledge and communication where it unfolds its full potential in the form of molecular animations. Molecular animation can be described as motion design for biological macromolecules. Since these nano-scale characters are not directly visible to the human eye, we need to draw on an array of visualisation methods to communicate them. Combining scientific illustration with motion gives us the opportunity to visualise the dynamics of the molecular system.Molecular animations can be used in science communication, education and research  and 2015In 2008 Gael McGill announced “Molecular Movies … Coming to a Lecture near You” , describThis perspective article formulates a number of guidelines with relevance for molecular animations based on knowledge and literature from the main fields design, scientific visualisation, molecular kinetics/kinematics and cinematography/storytelling. It may contribute to a theoretical basis for the field of scientific and especially molecular animation.Molecular animation is understood here as the visualisation of the structure and dynamic of macromolecular biomolecules and their substrates within the context of the living cell, at the molecular nano-scale. Molecular animation therefore can be seen as a subspace of data visualisation  and medical illustration/animation, that includes the biological mesoscale , methods (rising action), results (climax), analysis  and discussion (resolution) . In an aThe cinematic genre of molecular animations is best described as non-fictional documentary. Documentary storytellers must not invent and cannot make compromises when it comes to the facts. Instead, they need to be guided by and find the story in the material itself . The stoContextualised in the wider field of data visualisation, a molecular animation belongs to one of seven genres of narrative visualisation: film/video/animation. The narrative structure tactics is strongly author-driven. As such, an animation is characterised by linear ordering of scenes, heavy use of labels, headlines and annotations (messaging) and a lack of interactivity . The autAny story can be characterised by the three act structure that goes back to Aristotle’s Poetics  and FigMany molecular animation concepts include a storyboard with illustrated signature frames, the narration and possibly animatics for the timing. The more complex a topic and an animation are, the more important a storyboard becomes. Every signature frame, the transitions between them and the relative movement within a scene need to be designed to achieve the specific communication objective of that scene. Technically, this can be achieved by keyframe interpolation, particle or molecular dynamics with defined starting points and dynamic field parameters.The signature frames are individual images that represent important situations of the story. The molecular animator should be able to create a clear design for them and therefore have a good knowledge on design principles from various fields, most importantly graphic design, motion design, user interface design and cinematography/film. The design principles can be categorised into five  areas: draw attention, guide the eye, establish relationships, convey dynamics and create emotion /reaction. The methodology for the selection of principles relevant for molecular animations is described in the Supplement, including a visualisation of the principles in An animation is created by moving from signature frame to signature frame and includes the relative movement of characters and camera view. Characters may enter, stay in or leave the frame. Or they can move with the camera relative to other characters or the environment. Also, camera movement can be combined with character movement. The techniques and principles of non-dialogue cinematic storytelling  help to The motion lessons of interface design  support timing and spacing, easing, mass and weight, squash and stretch, follow through and overlapping, secondary action, arcs, solid drawing, anticipation, exaggeration, staging and appeal. With the help of those principles, natural movements are recreated based on material properties, following the laws of classical physics and building on every-day-life experience in the macro world. An experience that does not exist in the nano-world of atoms and molecules. Here, movement is determined by random collisions, diffusion gradients and thermal motion. So, with the exception of the more general principles staging and appeal, those for the design of motion need to be set aside for the animations of molecules.It needs to be mentioned, that the well known set of “Disney animation principles”  does notClarity is the overall goal of any design process and it is therefore also important for scientific visualisation in general. It is often reached by simplification (displaying fewer items) and abstraction (using simpler forms of an item), or both in combination. Clarity avoids clutter in the frame while unburdening the perceptual system of the viewer. However, it needs to be carefully balanced with the addition of more complex detail in order to advance the audience. In fact, in the nano-world of macromolecules, there is always a certain level of abstraction involved in scientific visualisations which automatically leaves room for interpretation . A taxonThe visualisation of a biological macromolecule (molecular graphics) can range from very simple to very complex. Illustrative and abstract representations include 1D formats (letter codes), 2D formats  and 3D formats . 3D-surface abstractions include beads representations  and coarse approximation. 3D atomistic surface models show more detail and include convolution surface models , molecular skin surfaces, ligand excluded surfaces, solvent excluded surfaces and solvent accessible surfaces. 3D atomistic space filling models are characterised by each atom being represented by a sphere with a radius based on the Van der Waals radii. 3D atomistic bond-centric models include hyperballs, licorice and ball-and-stick representations or even quantum mechanical models. Please see Choosing the adequate representation can be challenging and depends on several factors like the actual, specific communication objective, the experimental data available, the topical context, the target audience expertise level, and others. To avoid misconceptions, it is recommended to avoid causing superficial understanding due to over-simplification and abstraction. A depiction may be taken literally and not interpreted according to underlaying scientific knowledge. Whole biological concepts can be basically understood like this, but actual insight may not go beyond the simplicity of the representation explaining it . Mixed rAt the same time it should also be avoided to imply more knowledge than the data actually provides. The quality and resolution of the available experimental data restricts the level of structural detail shown in a scientific visualisation or animation. However, that does not keep us from integrating data from all kinds of resources or even models, predictions or hypotheses . Even wiThe design of motion requires biophysical knowledge from molecule kinetics and kinematics. For biological macromolecules four types of motion are most relevant: thermal motion, small and large conformational changes  and Brownian motion (with a molecule as one unit), see visualisations in Thermal motion is observed for any atom in a molecule and has a very high frequency, dependent on the temperature. The higher the temperature, the stronger the positional dislocation. It can often be neglected in an animation, especially when other, lower frequency motions are shown. It remains relevant in very complex and highly detailed animations, where visualising the movement of individual atoms or even quantum mechanical detail  increases the accuracy of the representation.Small conformational changes are based on the rotational freedom of a bond between two atoms, e.g., the rotation between peptide bonds see . AccumulBiological macromolecules and especially proteins often have domains, subdomains or other structural motifs  that show a certain rigidity within themselves while flexible loops between them are responsible for movements of those substructures relative to each other. Flexible loops themselves also may undergo large conformational changes to fulfil a function. Many biological processes depend on this type of large conformational changes. They are often visualised by the intrinsic movement of surface representations, but also ribbon-type cartoons.When macromolecules in a cellular environment move as one unit, they usually do so by random collision with other molecules, caused by their thermal motion and often described as random walk. Collisions create an external force that is not directional, so the movement of the macromolecule is random and not caused by long-range attracting forces between two reaction partners. This provides a challenge for a molecular animator, who needs to find a balance between the visualisation of the non-directional nature of the random motion and the actual approach of reaction partners within the timeframe of the scene. The motion-equivalent to structural detail in a representation is called dynamic realism by Jantzen . Both, sIn a crowded cellular environment, the set of principles described by In systems of high dynamics such as the nano-world of biological macromolecules, the medium of video can play out its strength. Compared to explanatory text, an animation can often be more efficient and intuitive. Compared to a diagram or illustration it can be more accurate and detailed. Consequently, an animation used in research and education should reflect the comprehensive knowledge of a system. Only then the complexity of the system can be communicated realistically and used for the development and evaluation of hypotheses. Failure to reflect the complexity leads to the misconception that a complex system is indeed simple and also to flawed future experiment design. As a consequence, it is recommended to use 2D sketches and cartoon style shading when little is known about a system and a 3D animation for established mechanisms .The main determinant for the complexity of a molecular animation is the complexity of the actual communication objective, the point that needs to come across, the focus of attention. The communication objective of a particular scene can be very specific and is usually related to one of these two categories (or a combination of both): a) the properties of the components - structure, chemical and physical properties, relation towards each other, etc.; and b) the dynamic of the system, the changes over time. The complexity of a communication objective is then directly associated to the complexity of the main element. The principle of consistent complexity  should be avoided. This is an issue for mixed representations. A more gradual change of the level of detail helps to avoid visual breaks. The environment should have less complexity than the characters, but may well become the focus of attention for another communication objective and have its complexity increased for another scene.The complexity of the structure representation needs to be matched with the one of the motion representation . While the complexity of a structure representation is easily understood as the level of detail, the complexity of a motion is less well intuitive. We need to look at the different motion types in order to relate the complexity of a representation with the complexity of a motion.


Ficus awkeotsang Makino extract (FAE) on Hs68 fibroblasts and BALB/c nude-mouse models are evaluated in this study. FAE was found to be non-toxic and showed high levels of DPPH, H2O2, and hydroxyl radical scavenging abilities; a ferrous chelating capacity; as well as ferric-reducing antioxidant capability. The antioxidant activity of FAE was strongly associated with polyphenolic content . The anti-inflammatory activity of FAE and the underlying molecular mechanisms were also investigated. The a* value of the mouse dorsal skin after treatment with FAE at 1.5 mg/mL in addition to chronic UVB exposure was found to decrease by 19.2% during a ten-week period. The anti-inflammatory effect of FAE was evidenced by the decreased accumulation of inflammatory cells and skin thickness. Expression levels of UVB-induced inflammatory proteins, including ROS, NF-κB, iNOS, COX-2, and IL-6, were significantly reduced upon FAE treatment in vitro and in vivo. Collectively, our results suggest that the inhibition of ROS and UVB-induced activation of the NF-κB downstream signaling pathway by FAE, indicating considerable potential as a versatile adjuvant against free radical damage in pharmaceutical applications.Antioxidant and anti-inflammatory activities of  The skin is the largest organ of the human body and the first line of defense against pathogens and environmental attacks . Solar uAcer nikoense and Betula platyphylla, was withdrawn from the market in 2013 due to cases of chemical leukoderma as a result of cytotoxicity induced by oxidative metabolites [Plant extracts often include ROS scavenger compounds that are effective in various ailments . Herbal abolites ,11. The Ficus awkeotsang Makino) is an indigenous vine species and a member of the Moraceae family. Jelly fig grows in the mountainous areas of Taiwan at 800 to 1800 m above the sea level. Numerous small achenes are obtained from the fruit of jelly fig after pollination and harvest, and pectin is extracted from dried seeds to produce jelly by mixing with water, which has become a popular traditional pectin-rich dessert for over 150 years in Asia [Ficus species, including fig (Ficus carica L.), Chinese banyan (Ficus microcarpa L.), and cluster fig (Ficus racemose L.), are excellent sources of antioxidants, whose free radical scavenging effects have been demonstrated previously [Ficus awkeotsang Makino extract (FAE) has an antioxidant activity. Hence, the aim of this study is to determine the antioxidant and anti-inflammatory effects of FAE, and the associated molecular mechanisms.Jelly fig  for achene production. The extract of Ficus awkeotsang Makino (FAE) was modified according to procedures previously described [escribed . Briefly2 and 37 °C. Cell viability was measured using 3--2,5-diphenyltetrazolium bromide (MTT)  after treatment with 0.25–2.0 mg/mL FAE, and also examined using a microplate reader at an excitation wavelength of 570 nm  according to the manufacturer’s instructions.Human skin fibroblasts (Hs68 cells)  were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and 100 U/mL penicillin/streptomycin at 5% CO2 solution. Then, the 5 mM ferrozine was added to the solution, and the mixture was shaken vigorously. The absorbance of the mixture was determined at 562 nm using an ELISA reader . EDTA was used as a positive control. The results were expressed as the percentage inhibition of the formation of the ferrozine-Fe2+ complex. The ferrous chelation activity was calculated using Equation (1) as follows:The chelation of ferrous ions was determined using the ferrozine assay with slight modifications, as previously described ,25. A se3Fe(CN)6 (1%) were mixed and incubated at 50 °C for 20 min. The reaction was stopped by adding 2.5 mL of 10% (w/v) trichloroacetic acid, and then centrifuged at 1000 g for 10 min. Finally, 2.5 mL of the supernatant was mixed with 2.5 mL of deionized water and 0.5 mL ferric chloride (0.1%). The absorbance of the reaction solution was determined at 700 nm using a microplate reader . Ascorbic acid and distilled water were used as the positive and negative controls, respectively. The reducing capability was calculated using Equation (2) as follows:The ferric-reducing antioxidant capability of FAE was measured using the potassium ferricyanide-ferric chloride method . Briefly2O2 in PBS. The resulting mixture was then incubated for 10 min in the dark at room temperature. Absorbance was determined at 230 nm using a microplate reader . The hydrogen peroxide scavenging activity was calculated using Equation (3) as follows:Hydrogen peroxide scavenging activity of FAE was measured spectrophotometrically, as previously described . FAE dis−1). The reaction was allowed to run for 30 min at room temperature in the dark, and the absorbance was measured using a microplate reader at 517 nm . Ascorbic acid was used as the positive control. The DPPH radical scavenging activity was calculated according to Equation (4) as follows:The α, α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging activity of FAE was measured according to a previously described method . Briefly3, KH2PO4-KOH buffer, H2O2, and ascorbic acid. Then, TCA and TBA were added, and the resulting mixture was incubated for 15 min at 100 °C. The absorbance of the mixture was determined at 532 nm using a microplate reader . The hydroxyl radical scavenging activity of FAE was reported as the percentage inhibition of deoxyribose degradation. The hydroxyl radical scavenging activity was calculated according to Equation (5) as follows:Hydroxyl radical scavenging activity of FAE was determined using a previously reported method . First, 2 UVB (emission peak at 302 nm) using a CL-1000 M UV crosslinker , and the intracellular oxidative stress of Hs68 cells was quantified using DCFDA fluorogenic dye . Fluorescence was detected using a microplate reader  at emission and excitation wavelengths of 520 and 488 nm, respectively. For immunofluorescence staining, HS68 cells were fixed on coverslips with 4% paraformaldehyde after exposure to 40 mJ/cm2 UVB. The slips were blocked with 5% skim milk in Tris-buffered saline (pH 7.6) containing 0.3% Triton X-100 and incubated with the following primary antibodies: anti-NFκB antibody  and anti-4, 6-diamidino-2-phenylindole (DAPI) antibody  for 30 min at 20 ± 1 °C. Following washing with PBS, the slips were incubated with the secondary-antibody anti-rabbit immunoglobulin (IgG)  for 30 min at 20 ± 1 °C. The cover slips were counterstained with ProLong Gold anti-fade reagent and DAPI , and images were captured using a fluorescence microscope .Cells covered with PBS were exposed to 40 mJ/cmRIPA lysis buffer  was used to extract total protein from Hs68 cells. The extraction was performed on a gradient sodium dodecyl sulfate (SDS)–sulfate-polyacrylamide gel, and transferred onto a polyvinylidene difluoride membrane. Membranes were blocked with 5% skim milk in Tris-buffered saline (pH 7.6) containing 0.3% Triton X-100, and incubated with the following primary antibodies: anti-cyclooxygenase-2 (COX-2) antibody , anti-inducible nitric oxide synthase (iNOS) antibody , and anti-actin antibody  at 20 ± 1 °C. Following washing with TBST, membranes were incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody  for 2 h at 20 ± 1 °C, followed by membrane incubation in an enhanced chemiluminescent (ECL) detection system  for target-antigen visualization. The intensity of the targeted protein bands was quantified and normalized to the actin expression area using the ImageJ 1.8.0v software .2 UVB irradiation 3 times a week for 1 min per exposure, and then treated with 100μL vehicle-polyethylene glycol (PEG) or 100 μL FAE on the exposure area per time for 10 weeks. Mice were divided into the following treatment groups: A, Normal group; B, UVB group; C, UVB + vehicle; D, UVB + 1.5 mg/mL FA extract; and E, UVB + 2.5 mg/mL FAE. The a* value was detected using a spectrophotometer  according to the formula provided by manufacturer’s instructions for 2, 4, 6, 8, and 10 weeks.The murine inflammatory assay was approved by the Institutional Animal Care and Use Committee of China Medical University . Five-week-old wild-type female BALB/c nude mice were purchased from the National Laboratory Animal Center , and acclimatized to the animal facility for one week. Mice were provided free access to water and food in polycarbonate shoebox cages with hardwood bedding at 21 ± 1 °C in a 12 h/12 h light/dark cycle. Mice were exposed to 180 mJ/cmHuman skin fibroblasts (Hs68 cells) were grown on a cover slip and treated with 0.25–2.0 mg/mL FAE after UVB irradiation. After 24 h, the cells were fixed with 4% paraformaldehyde and washed with PBS. They were then blocked with MPBS (containing 5% non-fat milk solution and 0.3% Triton X-100) for 30 min. PBS was again used to wash the coverslip, and incubation was performed with a primary antibody for 30 min and then with a secondary antibody, anti-rabbit immunoglobulin (IgG) . Coverslips were then washed with PBS to remove unbound secondary antibodies, and counterstained using ProLong Gold anti-fade reagent with 4′,6-diamidino-2-phenylindole. A fluorescence signal was observed using a confocal laser scanning microscope .−1) were mixed with 2.5 mL of 10% (w/v) Folin–Ciocalteu reagent and 2 mL of 75% Na2CO3 for 30 min at room temperature, followed by an absorption measurement at 765 nm using a microplate reader . Gallic acid was used as the standard to create a standard curve (0–100 μg mL−1), and the total phenol content was examined as mg of gallic acid equivalents of dry plant extract (mg GAE g−1 DW). Flavonoid content was measured as previously described [w/v) AlCl3-methanol solution, 0.2 mL of 1 M potassium acetate, and 5.6 mL distilled water were added to 1 mL of FAE solution (0.25–2.0 mg mL−1). The resulting mixture was incubated for 30 min at room temperature, and the absorbance was measured at 405 nm. Quercetin was used as the standard to prepare a standard curve (0–100 μg mL−1) to obtain mg quercetin acid equivalents of dry plant extract (mg QE g−1DW).Total phenol content was determined using the modified Folin–Ciocalteu method . Brieflyescribed . Briefly1H. Chemical shifts (δ) were expressed in ppm. Tetramethylsilane  was used as an internal standard [Nuclear magnetic resonance (NMR) spectral analysis was performed to determine the structure of the jelly fig extract using a Varian Unity Inova 500 MHz spectrometer  at 6250 Hz for standard . A high-n = 3). Statistical analyses were performed using the SAS-EG 7.1 software . One-way ANOVA and Tukey’s post hoc test were used to determine statistically significant differences  between data obtained from the control and test samples. Error bars were used to denote standard deviations from the mean (SD).Each experiment was repeated at least three times (p < 0.001) increased the viability of Hs68 cells, demonstrating the biocompatibility of FAE.Human skin fibroblast (Hs68 cell) is a standard cellular model of the human skin layer; it was used to determine if FAE could enhance the protective effect after UVB irradiation in a condition that closely resembled the physiological conditions. First, we determined the cytotoxicity of FAE in the Hs68 cell for further application. The viability of Hs68 cells following FAE application was evaluated using the standard MTT proliferation assay at 37 °C for 24 h . The low2O2 scavenging activity were found to increase by 27.1–82.7%  in iROS level in UVB-irradiated Hs68 cells was observed compared to the control. However, the level of change in the iROS level was decreased to 1.55 fold, 1.53 fold, and 1.74 fold, upon FAE treatment at 0.25, 0.375, and 0.5 mg/mL, respectively (The fold change in intracellular reactive oxygen species (iROS) was evaluated after UV irradiation. A significant increase  after UVB irradiation were determined. Expression levels of DAPI were approximately the same in all treatments, while a clear decrease in NFκB expression level was observed following the addition of 0.25–0.5 mg/mL of FAE in the culture medium, when compared to the UVB-irradiated treatment only . We alsoWe investigated the effect of FAE on skin inflammation following chronic exposure to UVB irradiation for 10 weeks using a BALB/c nude-mouse model. Redness was induced by UVB irradiation in BALB/c nude mice when applied alone or in combination with PEG (vehicle) treatment . To furtp < 0.001) differences in the a* value ranging from 7.1% to 19.2% with that of the non-irradiation control during a four-to-ten-week period. Statistically significant (p < 0.001) levels of suppression  of the redness of murine skin were detected after 2.5 mg/mL FAE treatment after UVB irradiation during a four-to-ten-week period. However, no significant difference was observed in the a* value between 1.5 mg/mL and 2.5 mg/mL FAE treatment. These results indicate that FAE is an effective anti-inflammatory agent following skin injury induced by UVB irradiation. A relatively low FAE dose (1.5 mg/mL) was sufficient to inhibit the inflammatory response following UVB irradiation.We further assessed the redness response of the dorsal skin of nude mice by determining the a* value, which serves as an inflammatory biomarker . Continup < 0.001) by 60.9%, 181.5%, 137.6%, and 127.6%, respectively, compared to that of the non-irradiated control. However, thicknesses of skin layers showed no difference with respect to the non-irradiated control upon FAE treatment at 1.5 mg/mL and 2.5 mg/mL. Immunohistochemical localization analysis . We also determined the total flavonoid content using the modified AlCl3 method. The total flavonoids content was determined to be about 10.3 mg QE g−1 DW using a calibration curve (regression coefficient = 0.9963) constructed using quercetin. The amounts obtained using this curve were thus expressed in terms of quercetin equivalents (QEs) per gram of dry extract weight. The flavonoids/phenol ratio of FAE was found to be 0.1  and high-performance liquid chromatography (HPLC) were used to determine the primary chemical structure of FAE. As seen in Finally, nuclear magnetic resonance spectroscopy (2−), hydrogen peroxide (H2O2), and hydroxyl radicals (HO•), which are formed during mitochondrial oxidative metabolism as well as cellular immune responses. ROS have evolved as regulators of intracellular signaling pathways [Reactive oxygen species (ROS) include radical and non-radical oxygen species, such as superoxide anions , gallic acid (IC50: 20.86 μg/mL), and BHA (IC50: 6.42 μg/mL) [To date, there are no previously separated components for jelly fig extract. Therefore, we first performed a pilot test (data no showed) and the result showed that the bio-activities of individual separated compounds were weaker than those of total jelly fig extract (FAE). The biological activity may be contributed to by the mixture of FAE. However, several studies have shown that flavonoids and phenolic compounds present in plants mainly contribute to their antioxidant activities ,53,54. P studied ,59,60,612 μg/mL) ,63,64. Hp < 0.001) in the presence of FAE, compared to that obtained with the vehicle treatment or control after UVB irradiation. This finding is in good agreement with those of several previous studies, in which the antioxidant of plant extracts were also linked to the inhibition of inflammatory-related activity in skin [Inflammation is an innate immune response that can be triggered by a variety of homeostatic changes, such as autoimmune reactions, infection, injury, toxicity, and disease ,66. The  in skin ,72,73. FWe further identified the HPLC spectrum of FAE was similar to that of β-sitosterol or stigmasterol. Although the compounds of those peaks remain unclear and require further confirmation by LCMS, it is worth mentioning that similar anti-inflammatory action was displayed in phytosterols and FAE. The β-sitosterol or stigmasterol are bioactive phytosterols found in many plant species . β-sitosFicus awkeotsang extract. This was followed by the downregulation of the proinflammatory signal transduction pathway, eventually leading to an anti-inflammatory response in nude mice (Nuclear factor-κB (NF-κB) is a key transcription factor in inflammatory cascades due to its ability to induce transcription of proinflammatory genes . The cytude mice .To our knowledge, this is the first study to identify the major bioactive components of FAE responsible for antioxidative and anti-inflammatory activities. FAE also exhibited significant antioxidative and anti-inflammatory properties without any cytotoxicity. Our data demonstrated that FAE function as suppressors of UVB-induced inflammatory action by downregulating iROS and NF-κB. In summary, our results suggest that FAE is a non-toxic substitute for synthetic antioxidants used in food, cosmetics, and pharmaceutical applications.


Responsive materials, i.e., smart materials, have the ability to change their physical or chemical properties upon certain external signals. The development of nanofibrous halochromic materials, specifically combining the pH-sensitive functionality and unique nanofiber properties, could yield interesting new applications, especially when the common problem of dye leaching is successfully tackled. Therefore, in this article, we studied the fabrication process of polysaccharide-based halochromic nanofibrous materials by using a combination of various halochromic dyes  and cellulose acetate in a spinning solution using a one-pot strategy. The inhibition of leaching was addressed by using a complexing agent: poly-diallyl-dimethylammonium chloride (PDADMAC). The preparation of hybrid spinning solutions, their characterization, and ability to form continuous nanofibers were studied using a high production needle-less electrospinning system. The produced hybrid solutions and nanofibers were characterized, in terms of their rheological properties, chemical structure, morphology, and functionality. Fabricated nanofibrous halochromic structures show a clear color change upon exposure to different pH values, as well as the reduced leaching of dyes, upon the addition of a complexing agent. The leaching decreased by 61% in the case of bromocresol green, while, in the case of bromothymol blue and thymol blue, the leaching was reduced by 95 and 99%, respectively. In the last decade, responsive smart materials became intriguing because of the dynamic and mostly reversible changes they introduce as a result of an external stimuli. In this area of material research, electrospun nanofibers, with their unique characteristics, such as an extremely high specific surface area, high porosity, small pore size, and high absorbance capacity, occupy a prominent position . NanofibSmart materials often contain halochromic dyes that consist of a pH-sensitive chromophore, responding to environmental pH by a color change . DependiSchueren et al. used co-spinning to develop pH-sensitive polyamide 6.6 nanofibers for a limited range of pH values . The stuRecently, there are some reports on incorporating halochromic dyes into polysaccharide-based matrix , in order to improve the biological use of these smart sensor systems. In previous research, Kurečič et al. investigated a pH-responsive drug release mechanism for wound dressing applications combined with the pH-responsive dye . The stuIn this study, the development of pH-responsive polysaccharide-based hybrid nanofibers with reduced dye leaching is presented via the addition of the complexing agent, PDADMAC. Halochromic dyes were homogeneously integrated into the spinning solutions of different CA concentrations, i.e., anionic dyes: bromocresol green (BCG), bromothymol blue (BTB), and thymol blue (TB), to provide the required responsivity to pH change. Cellulose acetate , acetic acid (AcOH ≥ 99.8%), bromocresol green (BCG), bromothymol blue (BTB), thymol blue (TB), and poly-diallyldimethylammonium chloride  were all purchased from Sigma Aldrich, Germany. For all experiments, ultra-pure water was used, prepared from a Millipore water purification system . All buffer solutions  were purchased from Reagecon Diagnostics Ltd., Ireland.Homogeneous CA solutions were prepared at four different concentrations  by dissolving appropriate amounts of CA in 85% AcOH and mixing for 24 h. A total of 4 wt% of complexing agent (PDADMAC) and 0.3 wt% of halochromic dye were added to individual CA solutions to obtain hybrid spinning solutions by mixing until homogeneous mixtures were reached (abbreviated as R-P-X-Y). Different halochromic dyes  were utilized to obtain a range of pH values. The mixing of the hybrid solutions was achieved by using a mechanical mixer  at 300 rpm for 24 h. All the hybrid spinning solutions were characterized, regarding their viscosity, conductivity, and color changing properties. Reference hybrid spinning solutions without the complexing agent PDADMAC were also prepared, in order to compare the effect of complexing agent on spinnability and leaching properties (abbreviated as R-X-Y). To do so, CA was prepared at different concentration from 9 to 15 wt% in 85% AcOH by mixing for 24 h using a mechanical mixer  at 300 rpm, and 0.3 wt% halochromic dye was added to individual CA solutions.All the abbreviations for prepared hybrid solutions are provided in The rheology of reference and hybrid spinning solutions was investigated by using Anton Paar GmbH, Rheometer MCR302 (AT), with cylindrical measuring system from 0.001 to 1000 1/s shear rate at room temperature. The conductivity of the hybrid solutions was measured using a Mettler Toledo International Inc. conductometer (CH), accessorized with conductivity probe InLab 710 for highly acidic conditions.Electrospinning was performed using a pilot scale needle-less electrospinning apparatus , which allows the formation of nanofibers on a support material with 50 cm width and infinite length . In the electrospinning process, wired electrodes were used as both upper and lower electrode, and different process parameters were varied to optimize the electrospinning conditions. The applied voltage was altered from 50 to 75 kV in 5 kV increments. Electrode distance was also adjusted from 100 to 190 mm in 10 mm increments. For optimal electrospinning of the different hybrid solutions, 140 mm electrode distance and 65 kV voltage were chosen for the comparison between different concentrations of CA and addition of different dyes. The ambient temperature was 19.2 ± 0.9 °C, and the ambient humidity was 27 ± 2%. The electrospinning duration was optimized at 30 min to obtain adequate samples. Hybrid solutions were electrospun on aluminum foil, as a support material, with dimensions of 300 × 400 mm. All the abbreviations for produced nanofibers are provided in The morphology of the electrospun nanofibers was observed using a scanning electron microscope  at an accelerating voltage of 1 kV after sputter coating the samples with a thin layer of palladium using a Benchtop Turbo sputtering device . The diameter of the nanofibers was measured using Image J  software. −5 M. UV/VIS Spectrophotometer Agilent Cary 60  was used to scan the prepared solutions, in order to determine the unique wavelength maxima (λmax) values needed for further analysis regarding responsivity and leaching. In order to evaluate the interactions between the dyes and PDADMAC, pH-potentiometric titration in water was performed, in order to quantify the charge present in the solutions for pure dye and PDADMAC–dye mixtures. For potentiometric titrations, 10 mg of dyes were titrated individually, starting from acidic to alkaline, with 0.1 mol/L HCl and 0.1 mol/L KOH as titrants. Additionally, mixtures of 4 wt% PDADMAC (1.2 g) and 0.3 wt% (0.09 g) of each dye were individually prepared and titrated, following above procedure. A two-burette auto-titration unit T70 , and a glass pH electrode DG-111 SC  was used to measure the pH of the solution continuously. Purging with nitrogen gas ensured an inert atmosphere during the measurements. The ionic strength was set to 0.1 mol/L (adjusted by the addition of 3 mol/L KCl). The pKa values of each dye and mixtures were calculated and presented. Data analysis and calculations were performed according to procedures described elsewhere . AdditioMonitoring of the responsivity of the electrospun nanofibers was performed with several buffer solutions with different pH values. Considering the color change of the halochromic dyes, which was shown in The color of samples was evaluated, in terms of the CIE L*a*b* color system, where L*, a*, and b* were the coordinates of the color in the mathematical combination of a Cartesian and cylindrical coordinate system, based on the theory that color is perceived as L* , a* (green—negative axis and red—positive axis), and b* (blue—negative axis and yellow—positive axis). The measurements were performed within a spectral range of 400–700 nm wavelengths using a two-ray spectrophotometer Spectraflash SF600 Plus  at standard illuminant D65 , from which the CIE color values were calculated using the Datacolor Match Textile 2.6.3.19R software  ,32. A XeExtent of dye leaching from each nanofibrous mat sample was analyzed using UV/VIS Spectrophotometer Agilent Cary 60 . Specified amounts of nanofibers (0.003 g) was immersed in separate 1× PBS solutions (5 mL), and leaching of the dyes was investigated by monitoring increase in absorbance at maxima wavelengths, characteristic for each respective dye. A single read UV/VIS measurement was performed at the beginning and end of 24 h immersion. For the successful fabrication of nanofibrous materials using the electrospinning process, it is essential to evaluate and optimize the spinning solution parameters according to their viscosity and conductivity. With optimal spinning solution parameters in place, formation of smooth, uniform, and homogeneous nanofibers can be ensured . There aIn In the case of measured conductivity values, presented in Morphology is an important characteristic of nanofibers, closely connected with their function, which is, in turn, connected to the active surface area. The morphology can be affected by the solution’s properties, as well as the process and ambient conditions ,53,54. DThe addition of PDADMAC to the reference hybrid solutions increased the conductivity of the samples and improved the electrospinning ability. As mentioned before, highly conductive electrospinning solutions produce nanofibers with smaller diameter size, along with the production of disc-like particles. On the contrary, the low conductivity of the reference hybrid solutions introduced a challenge for the electrospinning process. The effect of the viscosity and conductivity of the reference hybrid solutions results in small particle formation, along with challenges in electrospinning, as well as obtaining a significant amount of the sample ,56,57. FTo investigate the possible complexation between the components in hybrid solutions and nanofibers, halochromic dyes were titrated individually and in combination with the PDADMAC, in order to evaluate the pKa values. The acid dissociation constant (pKa) can be defined as the strength of an acid within different solutions . It can When titrated as mixtures (PDADMAC and dye), the pKa values of the mixtures change and shift towards higher values. It can be clearly seen that the charges present in pure dye and the mixtures have very different characteristics. During the titration process, along with pH increase, the negatively charged particles drastically increase for the prepared mixtures. It can also be concluded that, for the dye alone, it is a slow change, and the amount of negatively charged particles are low.From the calculations, it was found that the mixture of BCG and PDADMAC had 5.12 pKa, which indicates that an interaction between PDADMAC and BCG dye . The shiTo evaluate the responsivity of the dyes embedded in the nanofibers, a responsivity test was performed on electrospun nanofibrous mats. The nanofibers were exposed to buffer solutions with the selected pH values . The colors of the samples were also evaluated in terms of the CIE L*a*b* color system, where L*, a*, and b* were the coordinates of the color in the mathematical combination of a Cartesian and cylindrical coordinate system, based on the theory that was explained in the previous chapter. The CIE L*a*b* color space model was employed in this study for color evaluation, since it presents a good simulation of the human vision and is independent of the device used, as well as the nature of creation . As can As already pointed out, one of the main problems in integrating halochromic dyes into polymeric matrices is the management of leaching. The optical characteristics of the indicator dyes are very sensitive to their environment and depend on the way they are incorporated into the polymeric matrices ,64,65. VTo investigate the characteristics of the leaching, firstly, the pure dye solutions were analyzed. The dyes were dissolved in 1× PBS and scanned using UV/VIS spectrophotometer, from 200 to 700 nm, in order to identify their maximum absorption wavelengths. The 618, 430, and 435 nm wavelengths were determined as the unique signals for the dyes BCG, BTB, and TB, respectively, and they were used in the UV/VIS analysis. The nanofibers were peeled off and weighed (0.003 g) before they were separately immersed in 5 mL separate 1× PBS solutions. A UV/VIS measurement, at a specific dye absorption maxima wavelength, was performed at the beginning of the immersion and after 24 h of nanofiber immersion. A comparison between the nanofibers, with or without the addition of the complexing agent PDADMAC, were achieved, in terms of the 24 h leaching test. The results presented in The present study focused on the development of polysaccharide-based nanofibrous mats with sensing ability, which can find the applications in several fields, especially biomedicine. Halochromic dyes are often used as a sensing agent in smart, responsive materials, showing a wide range of color change. Due to the insufficient integration of halochromic dyes in the polymer matrix, the leaching problem is still a pronounced issue. In our study, we overcame this problem by introducing a complexing agent, PDADMAC, thus showing a significant reduction in dye leaching. The leaching of the dye was prevented up to 99% by the addition of the complexing agent, while maintaining the color changing ability of the halochromic dyes. The study also shows that the chemical structure of the dye has a great influence on the leaching mechanism. The introduction of PDADMAC does not disrupt the nanofiber formation; in fact, the addition of PDADMAC improved the spinning ability, as well as homogeneity, of the nanofiber’s morphology.


A macroscopic direct tensile test was employed to assess the polyurethane elastomers’ tensile properties. Our results show that the tensile strength of polyurethane elastomers increased with the increase in isocyanate content and decrease in temperature. Furthermore, the elongation at the break decreased, and the modulus increased, which is consistent with the law of molecular simulation.Mechanical properties determine the use of two-component polyurethane materials. The compatibility of two components in the polyether polyol-MDI molecular system greatly influences the formation of mechanical properties in polyurethane materials. In this paper, we studied and evaluated the compatibility and mechanical properties of two-component polyurethane at multiple scales by combining molecular dynamics simulation with macroscopic experiments, which is an important guideline for synthesizing and preparing two-component polyurethanes. We evaluated the stability of the two-component polyurethane system by calculating the solubility parameter, binding energy, and diffusion coefficient at four temperatures with three isocyanate contents. The Perl scripting language obtained the mechanical properties of the MDI-polyether polyol system. The MD calculation results show that the solubility parameter of two-component polyurethane negatively correlated with temperature, and the intermolecular binding energy and MDI diffusion coefficient positively correlated with temperature. When the mass ratio of polyether polyol to isocyanate was 1:0.6, the solubility parameter difference between the two was 1.43 (J/cm Polyurethane material is synthesized by the polycondensation of low-molecular-weight polyether/polyester and polyisocyanate. The main molecule chain comprises soft and hard segments. The different types and contents of two-component raw materials lead to differences in the number and structure of soft and hard chains, which causes the material to show various properties and mechanical characteristics ,2. ThereMechanical properties determine the use of polyurethane elastomers. However, due to the complex composition and reaction mechanism of two-component polyurethane, current research on the mechanical properties of polyurethane elastomers is mainly based on macroscopic experiments. Researchers often determine the effects of two-component content, additives, and the external environment on polyurethane elastomers’ mechanical properties by testing the mechanical properties of polyurethane elastomer specimens ,9. In adPrevious studies never predicted the mechanical properties of polyurethane materials in terms of the compatibility of the two-component blend. In addition, uniformly characterizing polyurethane is difficult on a molecular scale due to its complex composition. Performing molecular simulation calculations for two-component polyurethane is also difficult. Since polyether polyols and MDI are the main components of polyurethane elastomers, and the two are comparable on a nanoscale level, this paper selects the polyether polyol molecule–MDI molecular system as the research object. It uses Materials Studio molecular simulation technology and macroscopic experiments to study the two-component system at multiple scales. By analyzing the solubility parameters, binding energy, and molecular diffusion coefficient of the polyether polyol-MDI system at different temperatures and ratios, we judged the influence of temperature and ratio on the mixing effect of the two components. We used the Perl scripting language to calculate the mechanical properties of the two-component system and the relationship between stress and strain. Finally, we verified the microscopic mechanism obtained by molecular simulation using macroscopic experiments and analyzed the influence of temperature on the mechanical properties of two-component polyurethane molding specimens. These measures guide the formulation design of two-component polyurethane and the setting of each component parameter in the processing and production processes.t represents time, and ix, im, ia, and F represent the coordinates, mass, acceleration, and force of particles, respectively. In Formula (2), ix represents the coordinate of the particle, E represents the potential energy of the system, and the force F of the particle is equal to the first-order partial derivative of potential energy to the coordinate. MD simulation requires a rational selection of ensembles to reflect the actual physical process of materials. According to the different characteristics of the simulated object, the ensemble is divided into (1) micro-canonical ensemble (NVE); (2) canonical ensemble (NVT); (3) constant pressure, constant temperature (NPT); and (4) constant pressure, constant enthalpy (NPH). In the simulation of different ensembles, temperature and pressure balance control are usually needed to ensure that the simulation proceeds in the right direction. Equilibrium molecular dynamics calculates the molecular system optimization structure in different ensembles: energy, density, diffusion coefficient, heat capacity, etc. For the dynamic properties of the system, non-equilibrium molecular dynamics can be used to calculate the diffusion coefficient, thermal conductivity, viscosity, and other non-equilibrium properties of molecular systems.The basic principle of MD theory is solving Newton’s equation of motion. First set the motion of all particles in the system by following Newton’s equation of motion. The calculation formula isδ) is a physical index that predicts the compatibility between materials and is widely used in the field of polymer materials. Cohesive energy characterizes the cohesive properties of materials. Cohesive energy per unit volume is called cohesive energy density (CED). In polymers, the intermolecular force is a comprehensive reflection of various forces and cannot be expressed by a single force. Therefore, we used cohesive energy or cohesive energy density to measure the size of the force between macromolecules. In 1916, Hildebrand proposed the solubility parameter and defined it as the square root of the cohesive energy density of matter [E is cohesion energy (J/mol); V is the volume (mL/mol); E and V are the cumulative values of the energy and volume of the groups constituting the molecule; mU is the molar evaporation energy of the polymer; and mV is the molar volume of a repeating unit of the polymer.The solubility parameter (f matter .(3)δ= cannot be used as a sufficient criterion for blends’ compatibility; however, different binding energies between the blending systems can predict compatibility. The greater the binding energy, the more stable the formed group and the better the compatibility between polyether polyol and isocyanate. Therefore, the size of bindE is usually used to predict and compare the compatibility of similar systems. The binding energy (bindE) is the negative value of the interaction energy (interE), i.e., bindE = −interE.bindE and interE are the binding and interaction energies between the MDI-polyether polyol interface, respectively. totalE, poE, and MDIE are the average single-point energies of the two-component system, polyether polyol, and the MDI molecule, respectively.Intramolecular bonding interactions include bond stretching, bond angle bending, dihedral angle twisting, and off-plane bending. The role of intramolecular non-bonds includes Coulomb interaction and van der Waals force. Sometimes, intramolecular hydrogen bonds can also be used in intramolecular non-bond interactions. Binding energy (r(0) and r(t) are the atomistic positions of the center of mass at initial time 0 and later time t, respectively. The part in angular brackets denotes the mean square displacement (MSD) of all atoms.According to Brown’s law, molecules are always moving irregularly, and the mean square displacement (MSD) can characterize the activity of molecular chains in the model or the diffusion activity of particles in the system. The higher slope or shift in the MSD curve usually indicates higher molecular mobility, which benefits the effective motion of the two-component molecules. The mean square displacement represents the statistical square of displaced particles in the system relative to the initial time within a period of time. The formula isD can be obtained [D is diffusion coefficient; t is time; r(t) is the coordinate of the molecular; and k is the slope of the MSD curve.In order to quantitatively compare the diffusion ability of MDI molecules to different contents, the slope of the curve versus time is obtained by fitting the MSD curve, which is substituted into the Einstein relation for calculating the molecular diffusion coefficient, and the diffusion coefficient obtained :(7)D=limλ and μ) for isotropic materials are needed to fully describe their stress–strain behavior.λ and μ are Lamé constants; E is the Young’s modulus of the material; K is the bulk modulus of the material; and G is the shear modulus of the material.Mechanical properties are important properties in composite materials. In a molecular simulation, stress is generated inside the system under an external force so that the relative position of the particles changes. Only two independent coefficients (The molecular models of isocyanate and polyol are shown in We present three kinds of polyurethane research ratios in this paper. The mass ratios of polyether polyol (component A) to MDI (component B) were 1:0.5, 1:0.55, and 1:0.6, respectively. Then, we converted the mass ratio to the molar ratio in the amorphous cell section of MS software for two-component model construction. Under the COMPASS  force fi3, which is close to the real density of the material, indicating that the model built for this paper is close to the real state and is highly credible [credible .We conducted laboratory tests to evaluate the performance of polyurethane elastomer raw materials. The physical and chemical properties of each component are shown in We prepared three kinds of polyurethane elastomers by tensile testing. The two-component ratio is shown in The polyurethane elastomer tensile specimen size is shown in δ denotes the tensile strength of the polyurethane elastomer specimens (MPa); P is the ultimate load (N); and A is the initial cross-sectional area (mm2).L denotes the elongation at the break of the polyurethane elastomer test piece (%); L1 represents the distance between the marks when the specimen is broken (mm); and L0 denotes the distance between the front marks of the stretch (mm).The solubility parameter can be used as an index to judge the compatibility of two-component mixing . As shown in δ) between the two molecular models at different temperatures was generally minor, ranging from 1.2 to 1.7 (J/cm3)1/2. At 333 K, the two-component solubility parameter difference satisfies Δδ < 1.4–1.5 (J/cm3)1/2, indicating that the molecular system is the most stable. When the mass ratio of polyether polyol to isocyanate is 1:0.6, the solubility parameter difference between the two components is smaller than other ratios, indicating that the two-component system is more stable with increased isocyanate content. However, the difference in solubility parameters of the same two-component ratio at different temperatures did not show a certain rule. This finding is due to the different sizes and structures of polyether polyols and MDI molecules, resulting in different decrease rates of cohesive energy density and solubility parameters with temperature. It is also related to the physical properties of polyether polyols and MDI.According to the study, when the difference of solubility between polyol and MDI is less than two, the two can be compatible . As showWe calculated the total energy averages for the last 10 frames of the steady-state structures of the three two-component models with mass ratios of 1:0.5, 1:0.55, and 1:0.6 at four temperatures of 243, 258, 293, and 333 K using molecular dynamics. The results are shown in The greater the intermolecular interaction energy of the system, the stronger its intermolecular interaction ability and stability. When the interaction energy is positive, the molecules are mutually exclusive. When the interaction energy is negative, the molecules are attractive . We mainWe obtained the mean square displacement curves of the two components at different ratios using molecular dynamics simulation of the two-component miscible system. The results are shown in The diagram shows that the MSD values of each component under different proportions increase with simulation time. The MSD curve is approximately linear after stabilizing. The higher the temperature, the greater the value of MSD. The MSD value of isocyanate is greater than that of polyether polyol. We analyzed MSD images and found that the two-component MSD image was not a straight line. The curve’s trend is steep initially and tends to flatten at the latter position. Therefore, a straight line was selected as the research object when calculating the slope; moreover, the MSD of the linear interval was selected to calculate the diffusion coefficient. We repeatedly adjusted the range of values to find the most reasonable value.The diffusion coefficients of MDI and polyether polyol molecules at different two-component ratios are shown in In addition, with the increase in isocyanate content, the diffusion rate of MDI molecules gradually slowed down. When A:B = 1:0.6, the diffusion coefficient of the MDI molecule was the smallest because as the number of MDI molecules increases, the density of molecules in the system increases, and the free volume inside the material decreases, decreasing the jump probability of MDI molecules. On the other hand, with the increase in MDI molecular content, the interaction between MDI molecules is enhanced, which also reduces the jump diffusion of MDI between pores in the system.When using MD to calculate the mechanical properties of the system, the isocyanate-polyether-polyol molecular model is subjected to stress in all directions (six directions). The modulus also changes as the relative position of the particles in the system changes. When simulating the mechanical properties of the isocyanate-polyether polyol system, we set the simulation temperatures to 243, 258, 293, and 333 K. The modulus of each system was calculated by Formula (8). The MD simulation results are shown in E, K, and G decrease because, under the stimulation of thermal energy, when the molecule is disturbed, the vibration of the atoms in the system strengthens, and the atoms move in a large range. The slip between molecules increases, resulting in a decrease in modulus. However, as the isocyanate content increases, its mechanical properties significantly improve regardless of the temperature.The modulus of the two-component molecular model is shown in In summary, the polyurethane elastomer’s tensile strength and Young’s modulus increased with increased isocyanate content, whereas the elongation at the break decreased. This result is mainly because the hard segments of polyurethane elastomers are mainly provided by isocyanate. With the increase in hard segment content, the hard segments are connected, and the number of hydrogen bonds increases, leading to increased interaction and physical cross-linking between the segments . TherefoIn addition, with the decrease in temperature, the tensile strength and modulus of polyurethane elastomers increased, and the elongation at break decreased. This result is because the glass transition temperature of polyurethane elastomers is often lower than room temperature . The dec3)1/2, and they blend best. When the mass ratio of polyether polyol and MDI is 1:0.6, the solubility parameter difference between the two components is minor compared with other ratios, indicating that the two-component system is more stable with increased isocyanate content.The solubility parameters of polyether polyol and MDI decrease with the increase in temperature. When the temperature is 333 K, the solubility parameter difference between the two components is from 1.4 to 1.5 (J/cmThe increasing MDI content and temperature increase the binding energy between two-component molecules. When the temperature is 333 K, and the mass ratio of the two components is 1:0.6, the maximum value is 531.68 kcal/mol, and the stability of polyether polyol and the MDI system is optimal.As the temperature increases, the molecular motion in the system accelerates, and the diffusion coefficient continues to grow. The diffusion coefficient of MDI molecules decreases with the increase in MDI molecular content.The macroscopic tensile test results show that the isocyanate content and temperature change significantly affected the mechanical properties of polyurethane elastomers. With the increase in isocyanate content and the decrease in temperature, the tensile strength of polyurethane elastomers increased, the elongation at break decreased, and Young’s modulus increased, which is consistent with the law of molecular simulation.


CFI and ARMS2 genes can influence exudative age-related macular degeneration (nAMD) risk. The aim of this study was to assess the role of CFI rs10033900 and ARMS2 rs3750846 polymorphisms in susceptibility to nAMD for the first time in the Algerian population. A total of one hundred twenty four controls and seventy two nAMD cases were included in the present study. Genomic DNA was extracted from venous blood leukocytes. CFI rs10033900 and ARMS2 rs3750846 variants were determined by using the real‑time polymerase chain reaction method. Differences in allele and genotype distribution between the cases and controls were tested with adjustment for age by logistic regression analysis. A stratification of case and control groups by age (<65 or ≥65) and by gender  was also performed. Statistical analyses were done using SPSS21.0. No statistically significant association was observed between CFI rs10033900 and ARMS2 rs3750846 polymorphisms and nAMD risk . Stratification by age and gender did not show any significant association between these two polymorphisms and nAMD in a sample of the Algerian population. In our study, CFI rs10033900 and ARMS2 rs3750846 polymorphisms did not predispose alone to nAMD in our population. This study is a contribution to the enrichment of the bank data concerning the CFI and ARMS2 genes, reporting, for the first time, the allelic and genotypic frequencies of these genes polymorphisms characterizing the Algerian population. Increasing evidence shows that polymorphisms in  CFI), Complement Factor H (CFH), and Age-Related Maculopathy Susceptibility 2 gene (ARMS2) . Further, Studies, including Wang et al. study, demonstrated a significant association of chromosomal 10q26 genes, especially ARMS2, with AMD, suggesting this gene as the most likely major AMD susceptibility gene . ARMS2 p (p=0.8) , 24. In  (p=0.8) . Moreove (p=0.8) , 25. Als (p=0.8) . Further≤0.0006) . The preCFI and ARMS2 gene, reporting, for the first time, the allelic and genotypic frequencies of these genes polymorphisms characterizing the Algerian population.Finally, this study is a contribution to the enrichment of the bank data concerning the The authors report no conflict of interest.


Treatment intensification may reduce risks of recurrence and cancer-specific mortality, but it may increase adverse events and impair quality of life. However, optimal management for pN1 patients remains unclear. Nevertheless, few randomized control trials for pN1 are under investigation, and then more research is needed to establish an optimal therapeutic strategy for patients with pN1. This review summarizes current evidence on the treatments available for men with pN1, summarizes RCTs that included pN1 prostate cancer, and also discusses future perspectives.Pathological lymph node involvement (pN1) after a pelvic lymph node dissection represents one of the most unfavorable prognostic factors for disease recurrence and cancer-specific mortality in prostate cancer. However, optimal management for pN1 patients remains unclear. Thus, the guideline from the European Association of Urology recommends discussing three following management options with pN1 patients after an extended pelvic lymph node dissection, based on nodal involvement characteristics: (i) offer adjuvant androgen-deprivation therapy, (ii) offer adjuvant androgen-deprivation therapy with additional radiotherapy and (iii) offer observation (expectant management) to a patient with ≤2 nodes and a prostate-specific antigen <0.1 ng/mL. Treatment intensification may reduce risks of recurrence and cancer-specific mortality, but it may increase adverse events and impair quality of life. Few randomized control trials for pN1 are under investigation. In addition, there are limited reports on the quality of life and patient-reported outcomes in patients with pN1. Therefore, more research is needed to establish an optimal therapeutic strategy for patients with pN1. This review summarizes current evidence on the treatments available for men with pN1, summarizes randomized control trials that included pN1 prostate cancer, and discusses future perspectives. Pelvic lymph node dissection (PLND) is recommended during radical prostatectomy (RP) for prostate cancer in clinical practice, depending on risk classification . PLND isSo far, the only randomized clinical trial (RCT) performed for patients solely with pN1 prostate cancer showed that immediate androgen deprivation therapy (ADT) was associated with better overall survival (OS) than deferred ADT in patients with distant metastases or symptomatic recurrences . HoweverClinical evidence on optimal treatment for pN1 prostate cancer is limited, requiring the establishment of a treatment strategy since treatment outcomes for pN1 vary from satisfactory, with observation in patients with favorable characteristics, to unsatisfactory, even with intensive treatment in patients with unfavorable characteristics. This review summaries current evidence on the treatments available for men with pN1, RCTs that included pN1 prostate cancer, and discussed future perspectives.Several retrospective studies reported the prognosis in pN1 prostate cancer . Since tInterestingly, Moschini et al. have reported the natural history of clinical recurrence patterns of pN1 prostate cancer using a database from the Mayo Clinic, where almost all patients received adjuvant or salvage ADT, and 22% received adjuvant or salvage RT . In thatTreatment de-intensification is another approach for pN1 patients with a favorable prognosis. Tilki et al. had demonstrated that BCR-free survival and MFS were better in men who received adjuvant RT compared with no treatment or salvage RT using large case series data from Martini-Klinik . MoreoveRP with PLND could cause various postoperative complications such as urinary incontinence, erectile dysfunction, bladder neck contracture, and inguinal hernia, reducing the quality of life (QoL). The Prostate Testing for Cancer and Treatment (ProtecT) trial, which randomized patients to receive monitoring, RP or RT, found the most significant negative effect on urinary continence and sexual function in those patients undergoing RP . SimilarAdditionally, ADT can cause several adverse effects (AEs) including sexual dysfunction, hot flushes, bone fractures, metabolic effects, cardiovascular morbidity, fatigue, and neurological disorders . A prospSimilarly, salvage RT is also associated with toxicity. A prospective study evaluating salvage RT plus ADT after RP showed increased bowel dysfunction and urinary dysfunction by the end of RT. These rates improved after RT completion, but not completely. Meanwhile, erectile function presented no change during RT but showed an abrupt decline after RP . SimilarBased on this evidence, treatment addition after RP may lead to increased toxicity and reduced QoL. Meanwhile, treatment addition may reduce or delay recurrence, which may lead to a recovery of QoL by avoiding continuous ADT. However, there is little data on QoL and outcomes reported by patients with pN1. Thus, a prospective study on treatment strategies would be necessary.So far, various RCTs have been conducted to develop adjuvant therapy for high risk of recurrence and salvage therapy for BCR after RP. Due to the unfavorable prognosis among men with pN1 prostate cancer, the development of novel treatment to suppress recurrence and improve survival is necessary. However, the frequency of pN1 prostate cancer is low, and so far, only the RCT by Messing et al. enrolled patients with pN1 only . MoreoveTo date (31 March 2022), several RCTs recruiting patients with pN1 are being performed . Among tMoreover, six RCTs recruiting pN1 in addition to pN0 or pNx are under investigation . HoweverIn the future, ongoing RCTs may change the current therapeutic landscape in pN1 prostate cancer. As examined in the PATRON NCT04557501) trial, PSMA PET/CT is a promising modality that has the potential to change the landscape of treatment strategies for prostate cancer. Recently, proPSMA Study Group Collaborators have reported that PSMA PET/CT showed higher accuracy for pelvic nodal metastases and distant metastases than traditional imaging modalities . Despite57501 triIn conclusion, pN1 represents an unfavorable characteristic in patients who underwent RP with PLND. Currently, a gold standard strategy for pN1 has not been established, and survival of patients with pN1 is not satisfactory. Thus, the development of novel treatments with more efficacy and less toxicity is an unmet need. Treatment intensification by adding novel antiandrogen as well as treatment guided by next-generation imaging are currently under investigation, and may become a standard of care in the future. However, a few RCTs are underway. In addition, as studies on QoL and patient-reported outcomes in patients with pN1 are scarce, more research on QoL and patient-reported outcomes is needed to develop a better therapeutic strategy for pN1 patients.


Escherichia coli and Klebsiella pneumoniae accounted for more than 50% of deaths attributable to and associated with AMR, and resistance was high among multiple types of antibiotic class, including fluoroquinolones, carbapenems, and third-generation cephalosporins. There were 2 pathogen-drug combinations that caused more than 6000 resistance-attributable deaths: third-generation cephalosporin-resistant Escherichia coli and fluoroquinolone-resistant Escherichia coli. Conclusions: AMR in UTI is an unignorable health problem, both for the management of urology disease and for global antibiotic resistance. Special tailored strategies, including enhanced surveillance and rational use of antibiotics, should be developed for different regions according to the region-specific pathogen-antibiotic situations and resources.Background: There are still no detailed data about the burden of bacterial antimicrobial resistance (AMR) in urinary tract infections (UTI). Concrete knowledge of global and regional bacterial AMR data is crucial for developing informed programs and policies to control bacterial AMR and for prudent use of antibiotics to optimize antibiotic therapy in patients with UTI. This study aimed to provide comprehensive global and regional estimates for the AMR burden of UTI in 2019. Methods: Data were obtained from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD), including death, disability-adjusted life-years (DALYs), year lived with disability (YLD), and years of life lost (YLL) for bacterial AMR in UTI for 7 GBD super-regions, 21 regions, 14 pathogens, 13 antibiotic classes, and 66 pathogen-antibiotic combinations in 2019. The estimates were based on two counterfactual scenarios: drug-susceptible infection and no infection. Results: Globally, there were 64.89 thousand deaths  attributed to and 0.26 million deaths (95% UI: 0.18–0.36) associated with bacterial AMR in UTI in 2019. Among regions, the all-age death rates were higher in southern Latin America, tropical Latin America, and Europe and lower in sub-Saharan Africa.  Bacterial antimicrobial resistance (AMR) is a major threat to human health and well-being worldwide that requires global and urgent action to address ,2. It caUrinary tract infections (UTI) are among the most common bacterial infections leading to clinic visits and can cause serious sequelae, such as renal damage and urosepsis ,7,8. In Concrete knowledge of regional antibiotic resistance data is crucial for developing informed and location-specific programs and policies to control bacterial AMR and for prudent use of antibiotics to optimize antibiotic therapy in patients with UTI. In this study, using data obtained from the updated GBD study, we comprehensively present the global, regional, pathogen-specific, and pathogen-antibiotic combination-specific burden of bacterial AMR in UTI.In GBD 2019, UTI was defined as kidney infection that can lead to systemic symptoms such as fever and weakness and can cause discomfort and difficulty with daily activities, including nephritis, cystitis, and urethritis. The ICD-10 codes for UTI are N10, N10.0, N10.9, N11, N11.0, N11.1, N11.8, N11.9, N12, N12.0, N12.9, N13.6, N15, N15.1, N15.8, N15.9, N16, N16.0–N16.5, N16.8, N30, N30.0–N30.3, N30.8–N30.9, N34, N34.0–N34–3, and N39.0  attributed to resistance and 72.12 million [95% UI: 53.06–96.02] associated with resistance). However, Central Europe, eastern Europe, and central Asia had the highest death rates . Similarly, South Asia had the highest DALY counts, and Central Europe, eastern Europe, and central Asia had the highest DALY rates .Among the 21 GBD regions, southern Latin America , and tropical Latin America  were the two regions with the highest death rates, for the rates both attributable to resistance and associated with resistance. Then, the ranking is followed by three European regions . The regions of sub-Saharan Africa, including southern sub-Saharan Africa, western sub-Saharan Africa, eastern sub-Saharan Africa, and central sub-Saharan Africa, had the lowest death rates A. In terEscherichia coli dominated the global AMR-related deaths in UTI, accounting for about 40%, of which 26.52 thousand [95% UI: 19.79–36.30]) deaths were attributable to AMR and 101.52 thousand [95% UI: 80.65–130.29] were related to AMR. Klebsiella pneumoniae was the second most common pathogen responsible for the AMR-related deaths in UTI, with more than 10% of deaths attributed to AMR (8.33 thousand [95% UI: 5.48–12.47]) and associated with AMR (27.68 thousand [95% UI: 19.01–40.31])  B. Similawith AMR .Escherichia coli and Klebsiella pneumoniae ranked first and second among almost all 21 GBD regions in terms of the fractions of AMR deaths caused by 14 pathogens  to the lowest in North Africa and Middle East (30.88% of deaths attributable to resistance and 30.62% of deaths associated with resistance)  . For the regions .Globally, four antibiotic classes were each responsible for more than 100 thousand deaths associated with AMR: fluoroquinolones (169.15 thousand [95% UI: 119.64–240.75]), third-generation cephalosporins (113.06 thousand [95% UI: 78.60–163.97]), aminopenicillin (107.72 thousand [95% UI: 84.82–139.29]), and β-lactam or β-lactamase inhibitors (104.54 thousand [95% UI: 76.74–144.36]). For deaths attributed to AMR, fluoroquinolones (21.12 thousand [95% UI: 14.14–31.53]) were still responsible for the most deaths in 2019, followed by carbapenems (21.12 thousand [95% UI: 14.14–31.53]) and third-generation cephalosporins (10.98 thousand [95% UI: 6.15–18.12]) C. The DAGlobally and in almost all 21 GBD regions, fluoroquinolones, carbapenems, and third-generation cephalosporins were the top three leading antibiotics attributed to AMR , but theEscherichia coli (6909 deaths) and fluoroquinolone-resistant Escherichia coli (6871 deaths). Four pathogen-drug combinations each caused between 3000 and 6000 resistance-attributable deaths: carbapenem-resistant Escherichia coli (3617 deaths), trimethoprim/sulfamethoxazole -resistant Escherichia coli (3453 deaths), fluoroquinolone-resistant other enterococci (3294 deaths), and fluoroquinolone-resistant Enterococcus faecium (3004 deaths)  among 21 GBD regions, and for the combination of fluoroquinolone-resistant Escherichia coli, it caused the most deaths in South Asia (2296 deaths) and East Asia (1054 deaths) among 21 GBD regions  . For the regions .Globally, 64.89 thousand deaths from AMR in UTI would be prevented if all drug-resistant UTIs were replaced by susceptible infections, and 0.26 million deaths from AMR in UTI would be avoided if all drug-resistant UTIs were replaced by non-infections in 2019. Based on the counterfactual scenario of susceptive infections, the number of deaths caused by AMR in UTI was almost the same as the sum of deaths caused by testicular cancer, urolithiasis, and other uncommon urology diseases in 2019 . Based oThe all-age death rates of AMR in UTI were high in some areas of Latin America and Europe and lowest in areas of sub-Saharan Africa. These findings differ from the results in terms of the AMR burden for all infectious syndromes, in which the highest death rates occurred in the four sub-Saharan Africa areas . The burEscherichia coli and Klebsiella pneumoniae dominated the deaths attributed to and associated with AMR in UTI. This result was consistent with the results reported in other studies [ studies ,19,20. W studies . MoreoveEscherichia coli and fluoroquinolone-resistant Escherichia coli are the two combinations causing the most deaths from UTI. Targeting measures should be employed to treat UTI caused by Escherichia coli using appropriate antibiotics. Noteworthily, Klebsiella pneumoniae was the second-leading urinary pathogen responsible for AMR-related deaths, yet the number of deaths caused by Klebsiella pneumoniae in combination with any antibiotic class was not high among all 66 bacteria-antibiotic combinations. In 2017, the WHO published a priority list of antibiotic-resistant pathogens that pose the greatest threat to human health to help in prioritizing the research and development of new and effective antibiotic treatments [Escherichia coli and third-generation cephalosporin-resistant Escherichia coli were included in the list, and these two combinations were identified with a critical priority, which is the highest priority in the list. However, fluoroquinolone-resistant Escherichia coli, the second-leading pathogen-antibiotic combination causing AMR-attributed death, was not included. Some countries also issued bacteria priority lists, such as the USA [The regional data about bacteria-antibiotic combinations causing AMR-attributed deaths is helpful for figuring out the most serious category of AMR in UTI and for selecting appropriate treatments in that region. For example, in South Asia and East Asia, third-generation cephalosporin-resistant eatments . Among t the USA  and Cana the USA . These lTo tackle the challenge of bacterial AMR, global actions have been taken such as the Global Action Plan on AMR  and the Another important aspect of controlling antibiotic resistance in UTI is the rational use of antibiotics. Antibiotic treatment is the basic component for UTI therapy  and is fSeveral limitations should be acknowledged. First, although GBD collaborations made extensive efforts to obtain data from multiple sources, data about AMR are sparse in some low-income and middle-income countries. To tackle this problem, several methodological assumptions were made by researchers, and the fits of modeled estimates with available data were validated. Second, as there exists a discrepancy in the diagnostic standards, detection methods, and supervision systems among different regions and countries, the quality of the data for different sources is uneven. Third, data on the UTI-related AMR burden of different age groups, genders, and infection sites are unavailable. UTIs such as lower urinary tract infections often occur in elders and females, data on the burden of UTI-related AMR in these high-risk populations at different infection sites can help in taking measures to control antibiotic resistance.Escherichia coli and Klebsiella pneumoniae play the dominant roles in AMR, and resistance is high among multiple classes of antibiotic including fluoroquinolones, carbapenems, and third-generation cephalosporins. Special tailored strategies including enhanced surveillance and rational use of antibiotics should be developed for different regions according to the region-specific pathogen-antibiotic situations and resources. Bacterial AMR in UTI is a health problem that cannot be overlooked in urology, especially in southern Latin America, tropical Latin America and Europe.