Turmeric has been widely studied as a color indicator for pH variations due to its halochromic properties. It has been tested in solution or included in some polymeric matrices. Some studies have demonstrated that its change in color is due to the tautomeric species of curcumin, and this property can be observed even if turmeric is assimilated in a film or nanofiber. Chitosan/polyethylene oxide (PEO) polymers have been tested in previous studies. Polyvinyl alcohol (PVA) nanofibers are used as potential carriers of drugs once they are insolubilized. The aim of this work is to cross-link PVA with citric acid (CA) to insolubilize the nanofibers and determine the effect on turmeric’s halochromic properties. The nanofibers were treated with a sodium hydroxide (NaOH) solution, and a chromatic study was undertaken to determine color change. The change in color was assessed by eye (subjective) and by spectroscopy (objective). The nanofibers were characterized, in addition to the colorimetric study, by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) as well. The results demonstrate how thermal treatment induces cross-linking between the nanofibers, allowing them to keep their shape once the NaOH solution is applied to them. The opposite effect (solubilization) can be observed for non-cross-linked (NCL) samples. Although the final color varied, the cross-linked (CL) nanofibers’ halochromic behavior was maintained. It was demonstrated that during cross-linking, ester groups are formed from the free carboxyl group in the cross-linked CA and the ketones present in the curcumin under acid conditions. So, CA acts as an acid catalyst to bond turmeric to the cross-linked PVA nanofibers. The substantial progress and development that society has undergone in recent decades has brought about a significant change in many people’s lifestyles. Nowadays, human beings are purchasing smaller, safer, cheaper, and more rapidly produced products. Nanotechnology is a technique capable of producing and manipulating matter at the nanometer scale to create new structures, materials, and products .Nanotechnology has been able to demonstrate that materials at the nanometer scale can offer different characteristics than when they are at a larger scale. For example, ceramic materials have high brittleness except when they are present as nanoparticles, when they can be malleable. When working at the nanometer scale, elements such as copper can form substances that turn from opaque to transparent, and gold can change from insoluble to soluble .The advance of nanotechnology has driven development and innovation in the textile industry, enabling the manufacture of smart and multifunctional textiles with applications in various areas of health, pharmaceuticals, fashion, sports, transportation, protection, and high-performance products. Smart textiles are able to detect an external stimulus, whether mechanical, thermal, chemical, magnetic, electrical, optical, or physiological, and respond to it. Functional fabrics are designed to exhibit functions in addition to their aesthetic properties .The modification of textiles at the nanoscale allows the implementation of new functionalities or improvements in their performance without overshadowing their initial characteristics of lightness, flexibility, comfort, and aesthetics. The new functions provided can include hydrophobicity ,4,5, oleIn the textile industry, nanotechnology has also made its presence felt in the production of fibers. One of the main objectives of chemical spinning is to produce fibers with the smallest possible cross-section. Based on this objective, electrospinning has emerged as one of the most promising technologies capable of producing fibers with a sub-micrometer or even nanometer cross-section. Moreover, electrospinning is also a simple and versatile technique capable of producing nanofibers with a multitude of morphologies using highly diverse materials, from polymeric to ceramic materials, for example. The technique is able to offer exceptional characteristics, such as a high surface-to-volume ratio, an ultrafine fibrous structure connected to itself at a multitude of points, and high porosity and lightness ,19,20,21Another feature of the electrospinning technique is the possibility of adding different compounds to nanofibers, both superficially adhered and encapsulated inside them. Due to the simplicity and versatility of the nanofiber morphologies that can be obtained, this technology has been implemented in many industrial sectors, including self-cleaning textiles with applications in medical devices and protective clothing ,22, agroHowever, most of the research carried out to date has focused on the technical aspects of nanofibers, and their aesthetic side has been completely overshadowed by their excellent and diverse qualities that provide great value across many different sectors. However, their aesthetic aspect can also provide technical value; several authors have used different halochromic compounds encapsulated inside electrospun nanofibers as sensors to detect pH changes in the environment. The incorporation of halochromic compounds into nanofibers involves a dyeing process since the principle of halochromism is based on a color change in the compound as a function of the pH in which it is found.Agarwal et al. made electrospun PA 6 nanofibers that could encapsulate different halochromic dyes, such as Phenol Red, Methyl Red, Bromothymol Blue, phenolphthalein, and Bromocresol Green, with the aim of being able to detect color changes from pH 1 to pH 10, in a study that also determined the effect of the compound on fiber morphology . Van derCurcuma longa, which possesses the ability to change its color due to its diketone groups being converted to the keto-enol form when exposed to alkaline conditions [Naturally occurring dye compounds that offer halochromic characteristics have been used by other authors due to their chemical composition, in addition to being renewable, non-toxic, non-polluting, safe and biocompatible. Devarayan and Kim prepared electrospun cellulose nanofibers containing a natural pigment extracted from red cabbage; the pigment’s anthocyanin content results in a color change across a pH range of 1 to 14. They suggest a possible application in medical monitoring of alcohol intake levels via a saliva test . Maftoonnditions   nanofibers incorporating turmeric as a coloring agent in the polymeric solution for future application as a pH sensor in the field of air filtration. PVA is characterized by being a hydrophilic, non-toxic, biodegradable, biocompatible, semicrystalline polymer, qualities that allow it to be used in a wide variety of sectors. However, it has low water stability, which limits its applicability; as a result, PVA is often treated to ensure prolonged stability and improve its performance . The metThe objective of this study was to determine if cross-linking between PVA and CA influences the ability of the halochromic sensor to detect pH changes and generate color changes in turmeric. The possibility of creating non-water-soluble halochromic PVA nanofibers will increase PVA’s field of application. Due to the possible presence of suspended substances in the turmeric dispersion, the dispersion used in the study was filtered to avoid alterations in the electrospinning process. Another unfiltered dispersion was used to evaluate whether there are differences after filtration in terms of coloration and detection of pH change.The nanofibers were composed of polyvinyl alcohol (PVA) Mw 61,000 g/mol with a degree of hydrolysis of 98.0–98.8 mol% at 99% ± 0.8 purity, purchased from Sigma-Aldrich ; turmeric, purchased from a local herbalist; and citric acid (CA) at 99% purity purchased from Sigma Aldrich. The PVA with CA and turmeric dispersion was prepared in distilled water.The vertical metal collector of the electrospinning system was covered with wax paper supplied by Bosque Verde . The halochromic effect was evaluated using a 10 g/L solution of sodium hydroxide (NaOH) supplied by Sigma Aldrich, which has a pH value of 11–12.w/v and CA at 10% w/v with respect to the weight of PVA and stirred with an electromagnetic stirrer at 80 °C until complete dissolution of the PVA.To produce the dispersion to be electrospun, the turmeric was first dispersed at 5 g/L in distilled water with electromagnetic stirring for 2 h. After this time, half of the prepared dispersion was filtered using a sintered glass fiber disc with a nominal pore diameter of 1.2 µm. Subsequently, in both turmeric dispersions (filtered and unfiltered). PVA was added at 9% To characterize the viscosity of the dispersions, a FUNGILAB Visco Elite R viscometer from Instrumentación Analítica  was used with an R3 spindle and a stirring speed of 200 RPM. The electrical conductivity was measured at a temperature of approximately 22 °C in both solutions with a Crison Basic 30 Conductimeter . Surface tension was measured with a KRÜSS K9 tensiometer . For pH measurement, a GLP 22 pH meter from CRISON  was used.The electrospinning process was carried out using a Spinbox instrument from Bioinicia . The instrument’s vertical metallic collector was covered with white waxed paper, which the nanofibers were deposited onto to facilitate their handling. A 16-gauge capillary was chosen for the extrusion of the solutions to avoid clogging by the non-dispersed compounds. The extrusion had the following characteristics: a rate of 0.8 mL/h, an applied voltage of 19 kV, a distance between the extruder electrode and the collector of 12 cm, and a process duration of 30 and 120 min. The temperature of the environment where the experiment was performed was 25 °C, with a relative humidity of 53%.To cross-link the nanofiber veils, a TCF 120 oven from ARGO LAB  was used at a temperature of 190 °C for 10 min.The first characterization of the nanofibrous veils was undertaken with the naked eye by 5 volunteers to determine whether there was coloration in the electrospun nanofibers and to determine which of them had a greater or lesser color tonality.° standard observer and the standard illuminant D65 [The color of the nanofiber veils was determined by using a MINOLITA CM-3600d  reflection spectrophotometer. UV energy was included. The measurements were made with the CIE-Lab 10nant D65 ; the valAs the electrospun samples were circular in shape see , the colThe samples were examined by electron microscopy using a field emission scanning electron microscope FESEM ULTRA 55. Each sample was placed on a surface and covered with a layer of gold and palladium using a sputter coater in order to make the samples conductive. The samples were analyzed with the appropriate magnification and with an acceleration voltage of 2 kV.−1 resolution.Fourier transform infrared (FTIR) spectra were recorded in order to characterize each fabric’s surface. A JASCO FT/IR-4700 type A spectrometer  with an ATR accessory was used to record 16 spectra with a 4 cmAs a starting basis for sample characterization, a single polyvinyl alcohol dilution was tested against filtered (PVA_F) and unfiltered (PVA_NF) turmeric dispersions.The dispersion was filtered through a sintered glass fiber disc to remove any turmeric particles larger than 1.2 µm that could interfere with the electrospinning process by obstructing the capillary exit or hindering the formation of the Taylor cone. The characterization demonstrated the changes that occurred in the PVA starting solution when citric acid and turmeric were added. It can be seen how the presence of these compounds increased the viscosity up to 192.70 cP in the case of the PVA_NF dispersion; however, when the dispersion was filtered, it had a value slightly lower than that of the starting solution, 129.02 cP. The large increase in the conductivity parameter also observed was due to the addition mainly of CA, since it dissociates into hydrogen ions and citrate ions. The surface tension decreased only slightly in both dispersions with respect to the PVA solution. However, there was a noticeable reduction in pH in the dispersions due to their CA content with respect to the neutral pH from the initial submission.In order to prove that, in addition to the filtration of the solution, there is also a difference in the color of the nanofiber mat due to the electrospinning time, two electrospinning process times were employed, 30 and 120 min; thus, four samples were obtained: the samples from the filtered (F) solution electrospun for 30 min (PVA_F_30) and 120 min (PVA_F_120) and the nanofibers produced from the non-filtered (NF) solution electrospun for 30 min (PVA_NF_30) and 120 min (PVA_NF_120). The visual appearance of the samples can be seen in The visual test was carried out by five volunteers. They were presented with the four samples prior to cross-linking and were asked to rank, from highest to lowest, which sample had the yellowest coloration following the incorporation of turmeric as a dye. All the volunteers, without knowing the order decided by the other volunteers or the conditions for the electrospinning, ordered the samples in the same way, as shown in Reflectance spectroscopy was carried out to determine the coloration of each sample accurately and quantitatively. Measurements of the chromatic coordinates of each sample were taken at four different positions.Due to the high solubility of PVA in water, a cross-linking process must be carried out to facilitate the handling and applicability of the electrospun veils. After cross-linking, a change in the color shade of the samples, including the waxed paper, can be observed. The CIE L*a*b* color coordinates of the electrospun samples after cross-linking are shown in The waxed paper was also subjected to the same cross-linking process to see if it undergoes alterations on its own.The results obtained from the measurements of the cross-linked nanofibers show chromatic differences in relation to the original samples. The values obtained for the color differences with respect to the base surface on which the solutions were electrospun, waxed paper, are shown. The brightness values were hardly altered after cross-linking.The value of 15.37 DE*ab obtained for the cross-linked waxed paper shows a notable color difference with respect to the original sample (0 DE*ab). The cross-linking of the unfiltered samples, PVA_NF_30_CL and PVA_NF_120_CL, hardly alters the initial coloration as the DE *ab values are similar to those before cross-linking. On the other hand, the color difference for the PVA_F_30_CL and PVA_F_120_CL nanofibers has increased with respect to the original samples, with values of 13.75 DE*ab and 14.55 DE*ab, respectively, obtained after cross-linking.In all cases, the samples show high luminosity values. It can be seen that the samples electrospun for 120 min both present a higher luminosity than the samples that have been electrospun using the same solution but for 30 min.After the halochromic test was performed on the electrospun samples, both thermally cross-linked and non-cross-linked, new chromatic values were obtained.The halochromic test was also performed on the electrospun samples that were cross-linked to obtain an insoluble PVA b. The vaThe conditions used for PVA electrospinning were defined based on the microscopy analysis to ensure that no bed or solvent presence was visible. When the citric acid and turmeric were included in the PVA solution, the parameters varied, and although, apparently, a Taylor cone was created, the nanofibers generated beds, which indicated that the conditions should be changed . HoweverOnce the nanofiber veils were obtained, a drop of NaOH solution was placed on the surface of the nanofibers and left to dry. This was done on both cross-linked (CL) and non-cross-linked (NCL) nanofibers. The veils were subsequently analyzed by SEM in order to determine the effect. As can be seen in −1 for the non-cross-linked (NCL) samples and represented by lines, whereas the samples with thermal treatment show a shift towards a higher wavelength centered around 3306 cm−1. When the effect of filtration is compared (filtered (F) versus non-filtered (NF)), it can be clearly seen that there is a slight difference, mainly observed in the region 800–500 cm−1, which corresponds to the ring and OH bands [The FTIR spectra for the different samples were analyzed; the most important findings are shown in OH bands . For theFor the electrospinning samples, the lowest value obtained is for the PVA_NF_30 sample, with a value of 94.63 L*, while the filtered sample with the same electrospinning process time, i.e., the PVA_F_30 sample, has a slightly higher brightness, with a value of 95.38 L*. The samples electrospun for a longer time, 120 min, have higher brightness values. The sample electrospun from the unfiltered solution (PVA_NF_120) has a value of 96.42 L*, while the filtered sample, PVA_F_120, has the highest brightness value, 97.60 L*.Therefore, with respect to the electrospun samples, it is possible to state that the filtration of the turmeric solution increases the luminosity of the nanofibrous mats, in addition to it being possible to observe that a longer electrospinning time, i.e., a higher deposition of nanofibers, is also a parameter that increases the luminosity value.Regarding the color differences (DE*ab) b with reThe results demonstrate in both cases the higher color difference, i.e., the higher presence of color, in the nanofibers when electrospinning takes place for longer process times, 120 min in this case. They also show a notable difference in coloration depending on whether the PVA solution has been filtered or not, with the presence of darker areas observed when the solutions have not been filtered b.In The electrospun nanofibers from the unfiltered turmeric solution have very different chromatic coordinates than those obtained from the filtered solution. The PVA_NF_30 nanofibers have coordinates of −3.72 a* and 15.07 b*, which reflect a noticeable yellow coloration due to their high value on the Y-axis. The PVA_NF_120 sample has coordinates of −2.92 a* and 19.36 b*, where the value of +b*, responsible for the yellow coloration, is higher than that for the rest of the samples, so this sample is the one with the greatest yellow coloration.The chromatic coordinates have demonstrated that a greater coloration is obtained for the nanofibers when the turmeric solution is not filtered, in addition to the fact that a more intense color is obtained with longer electrospinning times. These results, as could be expected, agree with the results obtained from the volunteers’ observations .The filtration of the solution did not make any difference to the appearance of the nanofibers; while the image for the non-filtered samples has some areas with a higher intensity of color see , the colFrom the data shown in The electrospun samples after cross-linking show slightly lower values in terms of brightness, as can be seen in the points plotted in From The +b* value of the heat-treated collector wax paper increased. Initially, it had a cold white shade, and hence its value of −5.70 b* is towards the bluish area of the CIE L*a*b* space. After cross-linking, it had a yellowish hue, as indicated by the 9.37 b* value.The electrospun samples from the filtered solution (PVA_F_30_CL and PVA_F_120_CL) underwent significant color changes after the cross-linking of the polymer. The original samples presented a* and b* values very close to the center of the axes (0.0), thus representing a whitish coloration. However, the location of the points plotted for the cross-linked samples in The visual analysis performed allowed the determination of the color change in the electrospun nanofibers after the halochromic test was performed on them. The data in Firstly, with respect to the brightness values, it can be seen in Therefore, it is possible to affirm that the color shade after the halochromic test is also influenced by the electrospinning process time and the filtration of the solution used. It is noticeable how, in both the filtered and unfiltered solutions, the PVA_NF_120_H and PVA_F_120_H samples, electrospun for 120 min, show a greater color difference after the test than the samples made employing a shorter process time. The unfiltered samples, PVA_NF_30_H and PVA_NF_120_H, show greater color differences with respect to the filtered samples and the collector wax paper.On the other hand, in Therefore, by means of the halochromic test, it has been possible to determine the existence of a color change from yellow to purple for the nanofibers electrospun from the PVA and turmeric solutions. It has also been possible to affirm that the color intensity ratio is maintained with respect to the electrospinning and filtration time of the solutions.−1). This is because the number of OH groups decreases when the curcumin moves from the keto to the enol form, and consequently, this is directly related to the color change from yellow (pH ≤ 7) to a reddish hue (pH > 7), as many authors have reported [The curcumin in turmeric varies from a keto to an enol tautomer depending on the pH. The enol form is present when it is dissolved in organic solvents or alkaline solutions, whereas the keto form dominates in acid media ,40. Thisreported ,40.In the previous sections, the effect of thermal cross-linking on the colorimetric values of the dyed nanofibers has been confirmed; similarly, the color change from yellow to purple after the halochromic test has also been demonstrated. Therefore, in this section, we analyze both of the factors that induce changes; i.e., we evaluate whether the thermal treatment to which the nanofibrous veils are subjected to produce a water-insoluble polymer affects the halochromic capacity generated by the encapsulation of turmeric inside the fibers. The data shown in The electrospun nanofibers from the filtered turmeric solution hardly underwent a color change when the halochromic test was performed after cross-linking. As shown in −) at pH > 7 is reduced. This could be the reason for the color change associated with the halochromic effect when the nanofibers are cross-linked (CL) in comparison to the non-cross-linked ones.The cross-linking between PVA and CA takes place following the reaction shown in −1 for non-cross-linked samples (NCL) to 3306 cm−1 when the treated (CL) samples were analyzed. This indicates that OH groups were involved in the cross-linking reaction [−1. The SEM images show how a drop of alkaline liquid can dissolve the NCL samples, whereas the CL samples remain unaltered. This suggests that the thermal treatment enables the cross-linking of CA with PVA and converts soluble nanofibers into insoluble ones. This thermal procedure increases the applications of PVA as it can now be used in applications in which previously its high solubility would have prevented this use. Furthermore, CA acts by bonding the turmeric within the PVA-CA structure.Samples with thermal treatment show a cross-linked (CL) effect, as can be observed in the FTIR spectra . The OH reaction ,42,43. TThe use of turmeric as a natural pH indicator is well known; however, its halochromic behavior when incorporated into PVA nanofibers insolubilized by cross-linking with CA has not previously been demonstrated.SEM microscopy showed the presence of PVA nanofibers, and their appearance, apart from some beds, was not altered by the inclusion of CA or turmeric in the solution. This means that both the turmeric and CA were completely dissolved. When the solution was filtered, no variations were observed either.The halochromic effect was determined subjectively by visual analysis and objectively by spectrometry. Both results allow us to conclude that the non-filtered samples show higher color intensity, even though some areas of darker color can be observed in the non-filtered (NF) samples.FTIR spectroscopy confirmed the tautomeric variation from the keto to the enol form when NaOH was added to the surface of the nanofiber veil. Furthermore, this technique demonstrated a cross-linking effect between CA and PVA.The cross-linking treatment modifies the halochromic response but does not cancel it out altogether. This is due to the ester formed between CA and the keto form of curcumin, and we can conclude that CA plays a double role, on one hand, cross-linking with the PVA nanofibers to make them insoluble, and on the other hand, acting as an acid catalyst allowing the keto form to react with CA and bonding it to the nanofibers.The advantage of this treatment is that the nanofibers maintain their structure even when in an alkaline environment, whereas the non-cross-linked nanofibers become completely dissolved.This response allows insoluble PVA nanofibers to be generated and used as a non-soluble sensor, opening the door to multiple applications, including applications in the food industry and medicine.In summary, we can highlight that CA acts as a catalyst, converting curcumin into the keto tautomeric form. This form is able to react with CA in an esterification reaction. Consequently, turmeric is entrapped by CA. The ester reaction binds some of the hydroxyl groups and thus reduces the number of O-groups, which causes the change in color. This implies a decrease in the color intensity, but the halochromic effect remains.


Background and aim: As an oncogenic long noncoding RNA, differentiation antagonizing non-protein coding RNA (DANCR) was identified in many kinds of cancers. However, the specific function of DANCR in melanoma remains unclear. Here, we aimed to clarify the role of DANCR played in melanoma progression and the underlying mechanisms.Methods: TCGA data base and patients' tissue samples were used to analyzed the function of DANCR in melanoma progression. Transwell assay was used to detect cell migration and tube formation assay was employed to assess the ability of angiogenesis. Western blot, qRT-PCR, ELISA and IHC assay were used to examine VEGFB expression and secrection. Luciferase assay verified the binding of DANCR and miRNA.Results: We found that the expression of DANCR was positively related to poor clinical prognosis of melanoma. DANCR knockdown suppressed melanoma progression with a more significant suppression in vivo compared with it in vitro. Further detection showed that beyond promoting proliferation, DANCR also enhanced angiogenesis via upregulating VEGFB. Mechanistic analysis revealed that DANCR upregulating VEGFB through sponging miR-5194, which negatively regulated VEGFB expression and secretion.Conclusion: We demonstrated a novel oncogenic role DANCR played in melanoma and suggested a new avenue for melanoma therapy by targeting the DANCR/miR-5194/VEGFB signaling. As the 7th most frequent malignant cancer in females and the 5th most frequent malignant cancer in males worldwide, melanoma, originating from skin melanocytes, is one of the most life-threatening tumors Increasing evidences underlined the important role of lncRNAs in modulating biological activities of melanoma cells, including but not limited to cell migration Emerging evidences exposed diverse molecular mechanisms of DANCR regulating target gene expression. Functioned as ceRNA, DANCR sponged with miR-335-5p to regulate ROCK1 expression, as well as invasion, migration and proliferation of cervical cancer cells Regarding these and the important role of DANCR in several tumorigenesis, in present study, we explored the function of DANCR in regulating melanoma progression as well as the underlying molecular mechanism.2 concentration of 5%. DANCR stably knockdown melanoma cells and the corresponding negative control cells were created using short hairpin (sh) RNA specifically targeting DANCR or non-specific control shRNA contained in the replication defective Lentivirus, and the cells were named as shDANCR or shNC cells respectively. Lipofectamine 2000 reagent  was employed to transfected small interfering (si) RNA to knocking down DANCR or overexpressing miR-5194. miR-5194 inhibitor, purchased from RiboBio , was transfected into cells according to manufactures protocel. Differently treated cells were seeded into 6 cm culture dish with 5×105 cell per dish. 24 h later, cells were washed with serum free medium (SFM) triply and cultured with SFM (5ml) for another 24 h. Then the medium was collected and centrifuged to collect the supernatants. The collected CM was used immediately or stored at -80°C until usage.Human melanoma cell clones, A375 and SK-MEL-28 cells, were bought from American Type Culture Collection , which were raised with DMEM supplemented with fetal bovine serum (10%) with a humidified atmosphere keeping the temperature of 37°C and COWith approval and supervision from the institutional review board and informed consents from all patients, we collected forty melanoma tissues from patients accepted treatment in the First Affiliated Hospital of Xi'an Jiaotong University. All patients participated in this study did not treat with any preoperative therapy, including chemotherapy, immunotherapy and radiotherapy, with their diagnoses determined according to the histopathological evidences.Fresh tissues were carefully washed with sterile PBS  and stored at -80°C immediately after surgery for subsequent RNA extraction. For IHC assay , the tissues were fixed with paraformaldehyde, embedded with paraffin and cut into sections with the thickness of 5 μm before staining with primary antibody against CD31 or VEGFB using the DAKO Autostainer Plus system. Pathologist double-blindly viewed and analyzed the sections under high power (× 400) field.Melanoma microarray data about RNA sequencing of melanoma samples was downloaded from TCGA Data portal. The data were normalized to Z score before statistical analysis.6) were subcutaneously injected into the right ventral of every mouse. All mice tested their body weight every 5 days. After 4 weeks, the mice were sacrificed and the tumors were surgically harvested previous to immediate storage at -80°C for extraction of total RNA and fixed with paraformaldehyde for IHC assay. All of the animal experiments were accomplished following the guide of the institutional review board of the first affiliated hospital of the Xi'an Jiaotong University.A total of 12 BALB/c-nude mice, half male and half female, were allocated to two groups and randomly digitized. When the mice were 5-week-old, shDANCR or shNC A375 cells  was used to extract total RNA before reverse transcription into cDNA with PrimeScriptTM RT reagent kit purchased from Takara . Next, CFX96 real-time PCR system  was used to examine the expression of interested targets with SYBR Green PCR Master Mix (Takara) and specific primers. GAPDH was employed as the internal control for normalization. Sequences of the primers for qRT-PCR are as following:DANCR: forward-5′-GCCACTATGTAGAGGGTTTC-3′ and reverse-5′-ACCTGCGCTAAGAACTGAGG-3′; VEGFB: forward-5'-ACCCCCAACCCTGATAAAAG-3' and reverse-5'-TCCTCATTTCCTCCATCTGC-3'; GAPDH: forward-5′-TCAAGGCTGAGAACGGGAAG-3′ and reverse-5′-TCGCCCCACTTGATTTTGGA-3′.The primer of miR‐5194 was purchased from RiboBio and used according to the protocol of the manufacture.Cold PBS washed cells two times previous to lysis with protease inhibitor added RIPA buffer containing Tris , NaCl , NP-40 (1%) and sodium deoxycholate (0.5%). Protein, 30 μg per sample, was separated with SDS-PAGE gel (10%) before blotting onto the nitrocellulose membranes. The membrane was incubated at room temperature with skim milk (5%) and washed with TBST washing buffer, before incubation with primary antibody against VEGFA  overnight at 4°C. After washing with TBST washing buffer, the membrane was incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature and visualized with Molecular Imagery ChemiDoe XRS System from Bio-Rad Laboratories.Cold PBS washed cells were trypsinized, resuspended in cold ethanol (70%) and stored at -20°C for more than 24 h. Then the cells were centrifuged to remove the ethanol and washed with cold PBS for two times before resuspension with PBS containing RNAse A (0.5 μg/ml) and propidium iodide (50 μg/ml). The resuspended cells were stored without light at room temperature for half an hour previous to subject to flow cytometry analysis.3) were seeded into every well of the 96-well plate and cultured for various times. Then cells were washed sterile PBS and cultured with MTT solution (0.5 mg/ml). Four hours later, medium was carefully moved and 150 μl DMSO was added to each well. After the crystals were fully dissolved, the plate was subjected to the microplate automatic reader with a wavelength for 490 nm to detect the absorbance .Differently treated cells  and stained with crystal violet solution (0.1%). Then the number of visible cell colonies in each well were examined.167 purchased from R&D Systems Inc.  to add to the CMs.Concentration of secreted VEGFB in differently treated cells were detected with Human VEGFB ELISA kit  using CM previously collected. To modify the concentration of VEGF in CMs, we used recombinant human VEGFB5 cell/ml were added to the upper chamber (300 μl). Differently treated CMs (1 ml) or differently treated melanoma cells (0.5×105) were added to the lower chamber as the chemoattractant. 16 hours later, cells in the upper chamber were fixed with paraformaldehyde (4%) and stained with crystal violet (0.1%) with carefully protecting cells in the lower surface of the chamber. After wiping the upper surface of the inserts with cotton swab, cells migrated to the lower surface of the inserts were counted.Transwell migration system was used with 8-μm-pore inserts as the upper chamber . SFM suspending HUVECs with a concentration of 3×105 per well) were added into the 24-well plate coated with matrigel. About 5 hours later, plate was taken out from the cell incubator to take photographs. Two branching points perfectly continuous tube was deemed as a tube.SFM or CM diluted SFM (1:1) suspended HUVECs , which were named as pGL3-DANCR and pGL3-mt. The constructed plasmids, presence or absence of miR-5194, were used to transfect cells with X-tremeGENE HP DNA transfection reagent purchased from Roche . Dual Luciferase Assay kit purchased from Promega  was used to perform the luciferase assay according to the manufacture's instruction. For each data point, cells seeded in 3 independent wells were detected.p value less than 0.05 was identified as statistically significant.Differences between 2 groups were analyzed using student's t-test with GraphPad Prism . Pearson's correlation and linear regression analysis were also performed with GraphPad. The First of all, we analyzed the microarray data profiling of human melanoma tissues from the TCGA database to identify the DANCR expression in melanoma patients. According to the TCGA database, high-DANCR-expression group in melanoma had poorer overall survival comparing to low-DANCR-expression group Figure A. FurtheData from TCGA database also revealed that higher level of DANCR expression was accompanied with higher rate of mitotic cells Figure A, which in vivo, we used the subcutaneous xenograft tumor model with shDANCR-A375 and shNC-A375 cells injected into 4-week-old nude mice. After 4 weeks raising, no significant difference was found in body weight between two groups of mice . All the mice were sacrificed and the tumors were harvested, which showed significantly reduced tumor weight and tumor volume in tumors constituted by shDANCR-A375 cells  from A375 and SK-MEL-28 cells with or without DANCR knockdown, which were put into the lower chamber of a transwell system with HUVECs in the upper inserts. Results in Figure Taken together, results in figure To clarify the potential angiogenic factors involved in DANCR modulating interaction of melanoma cell and HUVECs, we implemented qRT-PCR assay to profile the different expressed angiogenic factors in A375 cells with or without DANCR knockdown. As shown in Figure To verify the function of VEGFB in modulating DANCR-involved angiogenesis in melanoma, we investigated VEGFB expression in the subcutaneous xenograft tumors. Unsurprisingly, suppressed VEGFB expression was found in DANCR knockdown subcutaneous tumors Figure A and 5B.167 into the CM of shDANCR-melanoma cells. We found impaired HUVECs recruitment with CM from VEGFB downregulated shNC-melanoma cells. Consistently, VEGFB167 rescued the weakened HUVECs migration induced by shDANCR-melanoma cells  and starBase (https://starbase.sysu.edu.cn/starbase2/index.php) to analyze the bioinformation of the potential target of DANCR, which indicated that miR-5194 could bind to both DANCR and VEGFB  and found attenuated ability of promoting HUVECs tube formation in miR-5194-overexpressed (miR-5194-mimics) melanoma cells Figure A. FurtheIn this study, we analyzed the microarray data profiling from TCGA database and verified the tumor promoting role of lncRNA DANCR in melanoma progression. Further exploration revealed that except attenuating cell proliferation, downregulated DANCR also diminished angiogenesis of melanoma, which is an important way for supporting tumors with nutrients and oxygen. We further found that VEGFB played the key role in DANCR mediating angiogenesis. Mechanically, we confirmed that DANCR elevated VEGFB expression through directly binding and sponging miR-5194, inhibiting which could rescue DANCR knockdown inducing VEGFB suppression. In summary, our results suggested that DANCR facilitated VEGFB expression and the sequential angiogenesis to enhance melanoma progression through directly binding with miR-5194. Clarifying the function of DANCR in melanoma development may open a new avenue for melanoma therapy and broadened the understanding of DANCR in promoting cancer progression.in vitro, more significant efficiency of tumor suppression was found in the xenograft model. The pale appearance of xenograft indicated the angiogenic role of DANCR, which also contributed a lot in DANCR promoting melanoma progression.Angiogenesis, complex but highly controlled, is a process of blood vessels formation from the pre-existing vascular, which plays pivotal roles in tumor survival, development and metastasis Angiogenesis usually starts from sprouting from the existing vessels that keep quiescent without stimulation of pro-angiogenic factors, among which VEGFB is one of the most important one Preliminary studies showed that miR-5194 could work as a tumor suppressor in Glioblastoma In conclusion, our study revealed the promotional role DANCR played in melanoma progression and neovascularization in melanoma microenvironment. Mechanically, DANCR directly interacted with miR-5194 and sponged it to diminish the suppression of miR-5194 on VEGFB expression. Therefore, DANCR might be a novel target for improving melanoma treatment.Supplementary figures and tables.Click here for additional data file.


The UK Government’s 2015 Antimicrobial Resistance (AMR) Review , betweenThe Lancet estimated that 4.95 million deaths were associated globally with bacterial AMR in 2019. Of these, 1.27 million deaths–more than HIV/AIDS and malaria combined–were directly attributable to resistance [Lack of diagnostics, rapid or otherwise, also continues to challenge our understanding of the true global burden of AMR. A recent report from the Global Research on Antimicrobial Resistance (GRAM) project in sistance , and theTo date, investment in diagnostics to aid in mitigating AMR has largely focused on increasing laboratory surveillance capacity. While AMR surveillance data from LMICs remains limited, the Fleming Fund and other investors in laboratory capacity have helped to improve data availability . The effInvestment in diagnostic development for AMR remains substantially below that for new antimicrobials . FurtherMycobacterium tuberculosis mutations [Distinguishing between bacterial and viral infections is just one enabler of antimicrobial stewardship. Another is identifying the causative pathogen and understanding its resistance profile. Nucleic acid-based tests have revolutionized the field of diagnostics, enabling clinicians to identify pathogens and their resistance genes in a ‘plug and play’ system within one hour . Next-geutations . DevelopAMR may not be as palpable a pandemic as COVID-19, but with an annual death toll in the same order of magnitude, it represents a clear and present danger. The same impetus and enthusiasm for investment and prioritization of diagnostics to help end the COVID-19 pandemic needs to be applied to AMR. Without this, Jim O’Neill’s ambition  of using


Neurodegenerative disorders related to aging pose a significant global health challenge , a distinguished neuroscientist. While pursuing her graduate studies at the Johns Hopkins University School of Medicine during the early 1970s, Dr. Pert made a notable contribution by identifying for the first time an opioid receptor . Animal toxins are products of natural selection and they can be found in a broad range of organisms .Moreover, recent studies might have contributed to a better understanding of how neuropeptides and/or neuroreceptors influence synaptic plasticity, which is crucial for the development, learning and memory processes. Advances in research might have highlighted the role of them in neuroinflammatory processes, potentially providing insights into the mechanisms underlying neuroinflammatory diseases. New findings might have emerged regarding the role of neuropeptides and neuroreceptors in brain development, particularly during critical periods of growth and maturation. One example from our Topic is the significance of L-lactate in learning and memory which is substantial. Research involving rats revealed that introducing external L-lactate to the anterior cingulate cortex and hippocampus led to enhanced decision-making in the former and improved long-term memory formation in the latter. While the precise molecular mechanisms through which L-lactate delivers these advantageous outcomes are actively under scrutiny, this study uncovered the molecular signature from supplementing with L-lactate; that can trigger the upregulation of crucial controllers involved in mitochondrial biogenesis and the reinforcement of antioxidant defense . Of greater significance, the published study expands the shared understanding within the field regarding the molecular variety present in cerebellar cells. The second paper, aimed to explore the involvement of microglia in obsessive-compulsive disorder. It was created a pharmacological mouse model using the serotonin (5-HT) 1A/1B receptor agonist RU24969 to replicate the monoamine dysregulation observed in this condition. The authors assessed the changes in morphology and functionality of microglia within this model and they suggested new evidence points to a strong connection between microglial dysfunction and the manifestation of stereotypic behaviors. In their model, the Brain-derived neurotrophic factor (BDNF) might serve as a viable treatment approach for addressing these behaviors .Finally, two other articles from our Research Topic that can be mentioned is a new molecular discovery regarding cells from the cerebellum. It was described a novel molecular marker, Pou3f1, that delineates specific sets of glutamatergic cerebellar nuclear neurons (In summary, studying neuroreceptors and bringing new Research Topics like ours in honor of Dr. Candance Pert is pivotal for advancing our understanding of the nervous system's function, dysfunction, and potential therapeutic interventions across a wide spectrum of neurological and psychiatric conditions. Many pharmaceutical drugs, including those used to treat neurological and psychiatric disorders, target specific neuroreceptors. Understanding the structure and function of neuroreceptors assists in designing drugs with more effective physicochemical and biological properties. Importantly, as our understanding of neuroreceptors advances, personalized treatment approaches can be developed based on an individual's receptor profile. This can lead to more tailored and effective treatments for various neurological conditions.PZ: Writing—review and editing. JJ-S: Conceptualization, Writing—original draft, Writing—review and editing.


This retrospective study aimed to compare the outcomes of toric implantable collamer lens (TICL) surgery with those of implantable collamer lens (ICL) implantation combined with limbal relaxing incision (LRI) in patients with low myopia and astigmatism.A total of 40 eyes of 28 patients who underwent TICL implantation and 40 eyes of 27 patients who underwent ICL implantation combined with manually LRI between 2021 and 2022 were included. Primary outcomes were manifest sphere and cylinder, intraocular pressure, visual acuity, and astigmatism parameters at 1 day, 1 week, and 1, 3, and 6 months postoperatively.The two surgeries showed comparable effects on manifest sphere and cylinder, intraocular pressure, and visual acuity . Surgery-induced astigmatism (SIA) was maintained as stable in the TICL group , but was significantly reduced in the ICL/LRI group  from preoperative to postoperative 6 months. The TICL group displayed significantly higher SIA and correction index at postoperative 1, 3, and 6 months than the ICL/LRI group  vs., 1.17 , p = 0.010; CI: 0.98  vs. 0.80 , p = 0.018). No complications occurred during follow-up.The effects of ICL/LRI are comparable to those of TICL in correcting myopia. TICL implantation displays better astigmatism correction than ICL/LRI.The online version contains supplementary material available at 10.1186/s12886-023-02941-1. Various techniques have been developed to treat myopia and prevent wearing eyeglasses or contact lenses. An implantable collamer lens (ICL) was first successfully implanted into the eye of a 45-year-old female in 1949 , 3. TICLThe implantation procedures for ICL and TICL are identical except for one extra step in implanting TICL –6. In boLike arcuate keratectomy (AK), limbal relaxing incision (LRI) is one of the methods used to correct astigmatism , 9. In bSince TICL and ICL combined with LRI (ICL/LRI) can correct both myopia and astigmatism, it is interesting to compare results of the two operations. This study aimed to compare the outcomes of TICL and ICL/LRI in patients with low myopia and astigmatism.This study enrolled subjects diagnosed with myopia and astigmatism who underwent TICL surgery or ICL combined with LRI surgeries at Beijing Tongren Hospital between September 2021 and September 2022. Inclusion criteria were: (1) patients who decided to accept ICL surgery; (2) had astigmatism less than 2.0D; (3) had differences between the steepest meridian of astigmatism and the corneal K2 axis of less than 10°; (4) aged between 18 and 50 years; or (5) with regular astigmatism (1D-2.5D). Patients with cataract, glaucoma, or fundus diseases were excluded. Patients were randomly assigned to the ICL/LRI group or the TICL group. All surgeries were performed independently by the same team.The study followed the Declaration of Helsinki. The study protocol was reviewed and approved by the internal review board (IRB) of Beijing Tongren Hospital, and all patients provided signed informed consent at the time of surgery for their perioperative data to be recorded for subsequent study and publication.Patients remained in a sitting position under the slit lamp to mark the horizontal point of the cornea. The patients then moved onto the operating table and lay flat. The cornea was marked with a 1 mL needle tip to label the placement axis for TICL. The main 3-millimeter incision was made at 180° and a 1-millimeter side incision was made with an 15° incision knife (MANI Inc. Japan) at another angle to allow position adjusting for TICL. After implantation, the TICL cylinder axis was aligned with the axis of astigmatism.The TICL and water-filled IA were inserted through the main incision, and the implantable lens adjustment hook was rotated and moved the TICL through the side incision. After the anterior chamber was maintained and the four haptics of the TICL were moved under the iris, the wound was cleaned using sterile saline. Patients were then sent to the recovery room.www.lricalculator.com) to get the incision site and length. The surgically induced cylinder was set at 0.4D. The calculated results were recorded. The incision length was also calculated by the equation developed by the author. In the equation, astigmatism in D was multiplied by 100 and divided by 2. For example, if a patient needs to release 1.6D astigmatism, the astigmatism release arc length is 1.6*100/2 = 80°. The maximum is 90°, with 10° flexibility for the operation. The equation was shown to provide similar results as the online software.Patients’ optometric data were entered into the LRI surgical planning software  was used to cut into about 90% corneal depth. With the help of the auxiliary device, a vertical to the surface and curved incision can be made. The main incision with the desired length was made at the opposite side of the first LRI. The ICL and water-filled IA were inserted through the main incision, and the implantable lens adjustment hook also helped to move the ICL through the main incision. After the anterior chamber was maintained and the four haptics of the ICL were moved under the iris, the wound was cleaned using sterile saline. Patients were then sent to the recovery room.Patients were examined preoperatively and 1 day, 1 week, 1 month, 3 months, and 6 months after surgery. The same optometrist conducted the ophthalmic examination for the patient throughout the whole follow-up procedure. Manifest sphere, Manifest cylinder, intraocular pressure, visual acuity (logMAR), and astigmatism. The CI is the ratio of surgical-induced astigmatism (SIA) to target-induced astigmatism (TIA). A correction index (CI) value > 1 (< 1) means RA overcorrection (undercorrection).We used G*Power 3.1.9.7 software for power analysis with the following settings: MANOVA: Repeated measures, within-between interaction,α = 0.05, power = 0.8, group = 2, number of measurement = 5, and effect size = 0.4. The calculated sample size was 80.Continuous data with normal distribution are presented as mean ± standard deviation (SD) and analyzed by Student’s t test; Continuous data without normal distribution are presented as median  and analyzed by the Wilcoxon rank sum test; Categorical data are presented as n (%) and analyzed by the chi-squared test or Fisher exact test, as appropriate. Generalized estimating equations (GEE) with unstructured matrix were performed to evaluate the association between the surgery type and the postoperative performance of manifest sphere, manifest cylinder, intraocular pressure, and visual acuity. Stratified analyses were performed to observe the difference between the left and right eyes. Vector analysis within different time were using Friedman’s test and presented as median (IQR) because of data without normal distribution. All p values were two-sided and p < 0.05 was considered statistically significant. All statistical analyses were performed using the statistical software package SAS software version 9.4 .A total of 80 eyes of 55 patients were included. The average age was 28.36 ± 6.09, and women accounted for 70% of patients. The TICL group contained 40 eyes of 28 patients and the ICL/LRI group contained 40 eyes of 27 patients.Table Figure Table Supplementary Tables In the right eye Table , no signTable The present study showed comparable effects of myopia corrections in TICL and ICL/LRI surgeries in patients with low myopia, while TICL displayed better astigmatism correction compared to ICL/LRI in patients with low astigmatism.Vector analysis revealed the following information about outcomes. First, the differences between SIA and TIA revealed that neither TICL nor ICL/LRI achieves a perfect astigmatism correction. Second, the median of CI at all the observed time points in the TICL group remained stable over the course of 6 months, and had better CIs. Both TICL and ICL/LRI surgery can maintain its therapeutic effects within 6 months, although TICL displays better astigmatism correction than does ICL/LRI.The observed time points in the present study were from the first postoperative day to 6 months after surgery. Compared with other studies, the starting point of observation was much closer to the date of the operation –13. ThisMyopia and astigmatism need to be minimal to achieve sufficient visual acuity. These emmetropias can be corrected by various procedures besides wearing eyeglasses. For astigmatism, procedures such as arcuate keratotomy (AK), clear corneal incisions (CCIs) along the steep meridian, opposite CCIs, limbal relaxing incisions (LRI), photorefractive keratectomy (PRK), laser-assisted in situ keratomileusis (LASIK), and TICL can be applied –16. AlthGenerally, patients’ corneas will be cut three times in ICL/LRI operation. In the present study, patients with less than 10° difference between the steepest meridian of astigmatism and the corneal K2 axis were included. A 10° difference can be considered the same in ophthalmological surgery. With this requirement, one LRI could be used as the ICL insertion site in our study. This reduced the number of incisions to two rather than three, which shortened the time for surgery and recovery. Based on our observation data for 40 eyes undergoing ICL/LRI, this procedure is acceptable.The auxiliary device used in LRI is worthy of mention. Although anesthetic eye drops and an eyelid holder were used for the LRI operation, the eyeball is still able to move around consciously. An unfixed eyeball increases the complexity and difficulty of the surgery. However, with this auxiliary device, the surgeons can press the eyeball slightly to fix its location when making an incision. This prevents the unnecessary failure of LRI procedures. Besides, the curvature of LRI is important for reaching optimal results. With this patented device, the surgeons can easily make a smooth curved incision rather than a jagged or straight line. A smooth curved incision then guarantees that the tension on the surface of the cornea is relaxed evenly. This will lead to optimal results of LRI operations with few side effects.The present study has several limitations. First, the retrospective study design and single-center nature of the study does not allow generalization to other populations. Second, our study focused on patients with regular astigmatism and less than 10° difference between the steepest meridian of astigmatism and the corneal K2 axis. Patients who were excluded may have different results if applying the same procedure. Third, LRI is only good at adjusting minor astigmatism. Consequently, only patients with astigmatism less than 2.0D were included in the present study. Patients with a higher degree of astigmatism may not be able to be treated with ICL/LRI. TICL might be the only option for such patients. Fourth, the use of this old technique that has been largely substituted by femtosecond laser-assisted incisions. Lastly, the follow-up time was only 6 months, which may not have allowed observation of complications and the long-term effects of TICL and ICL/LRI.In patients with low myopia and low astigmatism, the ICL/LRI surgery has comparable effects to TICL in correcting myopia. TICL displays better astigmatism correction than ICL/LRI.Below is the link to the electronic supplementary material.Supplementary Material 1


This paper describes the design philosophy for our cloud-based virtual reality (VR) co-creation environment (CCE) for molecular modeling. Using interactive VR simulation can provide enhanced perspectives in molecular modeling for intuitive live demonstration and experimentation in the CCE. Then the use of the CCE can enhance knowledge creation by bringing people together to share and create ideas or knowledge that may not emerge otherwise. Our prototype CCE discussed here, which was developed to demonstrate our design philosophy, has already enabled multiple members to log in and touch virtual molecules running on a cloud server with no noticeable network latency via real-time artificial intelligence techniques. The CCE plays an essential role in the rational design of molecular robot parts, which consist of bio-molecules such as DNA and protein molecules. IllustrVirtual reality (VR) is one of the promising ways to solve the above issues by representing biomolecules as virtual objects, which are visible and touchable with the help of a VR headset with a hand motion recognition device . AccordiIn order to answer the above questions, we have developed a cloud-based VR co-creation environment in which multiple members can log in to the molecular simulation of virtual molecules running on a cloud server as if they are all working together in the same room . The envOne of the plausible applications of the CCE is a bio-molecular design system in which one can design useful molecular parts with virtual DNA and proteins in advance of actual wet experiments. Since all molecular interaction results simulated in the CCE are only hypothetical works, careful interpretation and investigation from various backgrounds including biochemistry, biophysics, molecular design, and simulation are needed.In this sense, the multiple-user environment is mandatory for the CCE.Due to the breadth of our CCE project, this paper is intended to be a position paper to explain our design philosophy, and it will then be followed by technical and experiment-based papers as each component is tested to evaluate the merit of our design choices. The organization of this paper is as follows. 2Rational molecular design has already played major roles in countless fields, for example, in medicine and material sciences . However2.1Enhanced perspectives can be provided by using VR in combination with an interactive simulation system , 6. In V2.2Knowledge creation is one of our objectives for developing the cloud VR CCE. The creation of knowledge by connecting users virtually has been studied previously using grid computing, especially for drug discovery and design . SimilarA cloud-based VR CCE can bring people together to share ideas or knowledge in a way that may not be or seem possible in everyday life. To give an example with respect to our own work, when most think about “molecular sciences” it may conjure an image of a group of experts in lab coats speaking in jargon and writing down complex chemical structures, like what is seen in movie laboratories. We believe that if molecular science appeared more approachable, it would allow for more people to participate and understand what is happening at the cutting edge of research.The game-like interactive nature of our co-creation platform makes it a candidate for encouraging wider participation, and it may help to simplify multi-disciplinary collaboration and allow more people of all ages to enter into or learn about molecular science . Althoug2.3Guided experimentation can also be important in accelerating and communicating findings , 18. GuiWe have developed the CCE platform including human-in-the-loop VR simulation, cloud-based VR implementation, and real-time artificial intelligence-controlled hand motion prediction from a scratch in order to deal with the molecular interaction among millions of atoms in VR. In our own prototype, guided experimentation is already essential in the parameter searches and tuning of coarse-grained molecular dynamics simulations. Because the spatial and temporal scales of coarse-grained simulations can vary widely, people often need to find new models and parameters that can reproduce natural phenomena, and initially, there is often no set definition to tune simulations automatically with algorithmic or AI methods .The cloud aspect of our work touches on many areas of study, such as cloud-based collaboration, VR, and remote operation. These technologies have a wide range of applications, and thus there are many groups working to provide solutions with one or more of these technologies. Cloud-based VR could be used for connecting people across society and sharing information with greater immersion . One metThese benefits could expand the possibilities open to people if they could share VR objects and touch them naturally without feeling a disconnect due to network latency. The following sections focus on these points from the viewpoint of VR molecular modeling and real-time inference and training for remote virtual hand operation.2.4Of the systems in Some of the other systems use VR , 32–34 a3Owing to the recent boom in cloud-based services, smaller and more cost-effective cloud solutions are appearing , 46, briThe scalability of the molecular simulation system is critical when designing molecular parts for molecular robots. Conventional molecular dynamic simulation targets from X-ray crystallography have molecular weights from tens to thousands of Daltons . In contConsequently, network performance is critical in cloud VR system development because poor network performance leads directly to a poor user experience . BandwidNetwork bandwidth affects how much data can be sent to the client and how frequently the data can be updated. Therefore, our bandwidth needs are partly tied to the simulation scale, but, in general, if there is more bandwidth available, the user has a smoother experience. There is a limit to the effect of the amount of available bandwidth because updating faster than the simulation rate or the refresh rate of the user’s screen/VR goggles is wasteful. An early evaluation of the VR cloud bandwidth performance is shown in Latency can create a time discrepancy between user actions and the presentation of the simulation results. If the latency is high, the user is more susceptible to suffering from VR sickness and the overall experience is poorer . Thus, n3.1To minimize network latency, we used our real-time inference and training to predict remote virtual hand motions . As we dIn order to touch a virtual object with a virtual hand in the VR simulation running on a remote server, the virtual hand must reach the virtual object before the user’s hand reaches the position of the virtual object to overcome the network latency issue. By having the future state (prediction) of the user, the cloud server begins to create an updated simulation state ahead of time so that even after the time required to create a new simulation state and send it to the user has passed, it is still up to date.We believe that this is a crucial technology that will help us to realize our CCE; the key to a CCE’s success is the user experience. If the interaction in the environment is not fluid like a real-life experience, users will likely leave the platform prior to even testing interactions or the simulation models that have been implemented. Consequently, this was our first priority in developing our CCE and the other components are now actively in development.In our past work on prediction , we pres4This section provides a complete overview of our current CCE prototype, which is a custom implementation that has been built by our group using languages and application programming interfaces, such as C++ and CUDA, for the simulation, and DirectX 12 for the user VR rendering.4.1Our CCE hardware consists of a central server, which in our case is in the cloud; however, it could also be a local server depending on users’ resources and security concerns. As for the users who connect to the cloud, their machines are responsible for rendering the simulation environment reported by the server. A visual overview with additional information is shown in 4.2In contrast to some game streaming services, such as GeForce Now or Google Stadia, and other visualization works , 53, we Some modern multiplayer games include dynamic environments that are simulated based on physical interactions; however, the degree to which the environment can be changed is still limited compared with reality . In addi4.3Currently, users can load VR molecular objects in Protein Data Bank format , and the. This allows us to probe systems live and interactively in any way that we choose to observe the resulting dynamics. The shared dynamic VR objects can be handled through the conventional wide area network or the Internet. This is one of the major differences between our approach and the game streaming or remote rendering approaches [For our prototype in its current state, the primary hypothesized benefit is the ability to share dynamic VR objects easilyproaches , 53, 55,4.4The most important next step for our CCE is to conduct user testing to verify the proposed benefits of our system and to receive user feedback for further improvements. To begin to accomplish, this we have already deployed our system with our collaborators at Hokkaido University and Kansai University, who work on molecular robot research in wet laboratories.Following initial user testing, we are also planning to expand the range of simulation models included within our CCE to enable more users to benefit from our system and to obtain more feedback for continuing to improve our platform. Part of expanding our user base will also include finalizing our mobile VR client application which will reduce the hardware requirements of users further.5In parallel to the development of our CCE, members of our team are working on additional VR environments to be added to our CCE to promote further research collaboration in the CCE. One of these environments is a VR viewer for microscope images. Microscopes, such as atomic force microscopes and confocal microscopes, are essential in the rational design of molecular robot parts . HoweverWe adopted a simple, effective visualization approach in our simulation. In confocal microscopy imaging , for a sCurrently, the method supports volumes with up to 512 × 512 × 512 resolution. 6COVID-19 has dramatically changed the world. One specific area of change that has touched the lives of nearly everyone is the shift in drug and vaccine design . SimilarMolecular robots  could beThe objective of our VR simulation is to observe the dynamics of a wide range of molecular interactions including DNA nanostructures and microtubule motion pattern formation to find a good initial distribution of atomic molecular models to which conventional molecular simulation can be applied. By simulating at atomic levels such as DNA nanostructures we aim to identify and understand the building blocks of molecular robots .Then from there, we can add the findings of such work into higher-level coarse-grained simulations focusing on molecular robots . Due to A molecular robot approach is superior to a mechanical nanorobot approach in terms of miniaturization because a molecular robot can use sources of chemical energy, such as adenosine triphosphate (ATP) , which iSeveral molecular robot prototypes have been reported, including self-propelled liposomes –40, muscAlthough much fundamental research has been reported in the field, practical applications of molecular robotics remain in the preliminary stages. To solve this problem, we have been developing a cloud-based CCE for rational molecular robot part design using VR , 4 and r6.1Our own collaborators and other teams around the world have found that a critical issue is the acceleration of the molecular part design cycle –44. MoleFor simulation technologies, there is also a lack of rational molecular design tools applicable to molecular robot parts, such as DNA origami, protein complexes, or microtubules. Consequently, conventional molecular dynamics simulations have been used, but these often take more than several weeks to run, even on high-performance computers . This haIn an effort to reduce some of the simulation challenges, previously we created a coarse-grained model utilizing GPGPU to accelerate the simulation of microtubule gliding assays , which a6.2To accelerate the molecular part design cycle, we have begun to develop a cloud-based virtual reality co-creation environment (CCE) augmented by real-time artificial intelligence (AI). The CCE aims to provide molecular objects and equipment in virtual reality as a service via the cloud so that researchers can collaborate remotely. To compensate for the latency caused by the distance between researchers, real-time AI technology, known as real-time inference and training (RTIT), has been implemented and plays an essential role in ensuring remote operation feels natural in the cloud-based virtual reality (VR) environment , 4, 51.7A CCE is similar to conventional CAD software, in that it could be used for designing beneficial or harmful things. Although this problem is difficult to address, CAD software has been accepted despite the fact that it can be used to design anything.The democratization of knowledge is another challenging question. It would likely be beneficial to educate more people with the aim of benefitting society as more people could then participate in industries that aim to solve future medical problems or create innovative materials. However, some people may have bad intentions, and limiting access based on intention is a challenging problem.The biological results that may be produced after the CCE is used for the design stage would need their own ethical consideration, which lies beyond the scope of this paper on the CCE. We are currently talking with ethics researchers about the innovative technologies in this project, such as molecular robotics .8By following the design philosophy described in


Contrastive learning has been widely used as pretext tasks for self-supervised pre-trained molecular representation learning models in AI-aided drug design and discovery. However, existing methods that generate molecular views by noise-adding operations for contrastive learning may face the semantic inconsistency problem, which leads to false positive pairs and consequently poor prediction performance.To address this problem, in this article, we first propose a semantic-invariant view generation method by properly breaking molecular graphs into fragment pairs. Then, we develop a Fragment-based Semantic-Invariant Contrastive Learning (FraSICL) model based on this view generation method for molecular property prediction. The FraSICL model consists of two branches to generate representations of views for contrastive learning, meanwhile a multi-view fusion and an auxiliary similarity loss are introduced to make better use of the information contained in different fragment-pair views. Extensive experiments on various benchmark datasets show that with the least number of pre-training samples, FraSICL can achieve state-of-the-art performance, compared with major existing counterpart models.https://github.com/ZiqiaoZhang/FraSICL.The code is publicly available at  As mostuch data . On the uch data . TherefoRecently, as self-supervised pre-trained models (PTMs)  encoder, while preserving the molecular structural information. A multi-view fusion mechanism is introduced to FraSICL to make better use of the information contained in different views. In addition, an auxiliary similarity loss is designed.Aiming at solving this semantic inconsistency problem, this paper proposes a Our contribution are summarized as follows:We raise the semantic inconsistency problem in molecular view construction for molecular constrastive learning and develop an effective method to generate semantic-invariant graph views by changing the graph topology while preserving the structural information of molecules.We propose a novel Fragment-based Semantic-Invariant Contrastive Learning molecular representation model for effective MPP, which is also equipped with a multi-view fusion mechanism and an auxiliary similarity loss to better leverage the information contained in unlabeled pre-training data.Extensive experiments show that compared with SOTA pre-trained MPP models, the proposed FraSICL can achieve better prediction accuracy on downstream target tasks with less amounts of unlabeled pre-training data.In this section, we first formally define semantic-invariant molecular view, then propose a semantic-invariant molecular view generation method and a multi-view fusion scheme. Finally, the structure of the Fragment-based Semantic-Invariant Contrastive Learning (FraSICL) molecular representation model and its loss functions are introduced.semantic-invariant molecular view.In the Section 1, we have given an example to illustrate how noise-adding operations may lead to semantic inconsistency and consequently false positive pairs. Here, we formally define m and its molecular graph G = V denotes the set of nodes that represent the atoms, E denotes the set of edges between nodes, representing the bonds. transformation function molecular graph view (or simply molecular view) G, i.e. Definition 1m is the same as the molecular graph semantic-conflict view of m with regard to (w.r.t.) (Semantic-conflict view) If there is another molecule Definition 2m is the same as a view semantic-ambiguity view of molecule m w.r.t. molecule (Semantic-ambiguity view) If there exists another molecule Given a molecule m, if it has a semantic-conflict view m and Both semantic-conflict views and semantic-ambiguity views will lead to false positive pairs for molecular representation contrastive learning. For example, assume that a GNN On the other hand, if a semantic-ambiguity view is generated as defined in Definition 2, i.e. Definition 3m with graph G, if semantic-invariant view of m.(Semantic-invariant view) Given a view To boost the performance of contrastive learning for MPP, we should avoid the generation of both semantic-conflict views and semantic-ambiguity views. That is, we generate only semantic-invariant views, which are defined as follows:According to Definition 3, semantic-invariant views should be neither semantic-conflict views nor semantic-ambiguity views. Besides, they should also be discriminative so that they can be encoded into different representations by neural network encoders. To achieve these goals, we propose a semantic-invariant view generation method.Although adding noise to molecules may lead to semantic inconsistency problems, it is still an efficient way to generate discriminative views for molecular graphs. Therefore, strategies to add noise should be carefully designed. Recently, researchers have discovered that graph rewiring, i.e. adding or removing bonds to or from the input graph to make the information propagating along a different topology inside a GNN encoder while preserving the original structural information by encodings , will ovm, its molecular graph can be denoted as an annotated graph E, we obtain fragment-pair view.Given a molecule Though this method seems similar to some other edge removal methods, this carefully designed strategy will bring advantages from the following perspectives:E, G. So the GNN encoder will generate a different embedding for By randomly removing one edge from edge set E is modified in i and j in graph i and j recorded in NOT a valid molecular graph of any molecule. Therefore, Since only the edge set ONE single bond is removed in view i and j, and the deviation is only 1. So, the removed edge can only be a single bond between i and j. Thus, there is no other molecular graph Since only Compared with randomly breaking one arbitrary bond of a molecular graph, our method can generate graph fragments that may correspond to some chemical meaningful functional groups, since acyclic single bonds can serve as boundaries between functional groups . And expm with molecular graph molecule view branch for generating molecular view, and the right one is the fragment view branch for generating the fragment view.With the proposed semantic-invariant view generation method, the structure of the FraSICL model is shown in MolEncoder, to capture the representation of the molecular graph In the molecule view branch, a GNN ntive FP  is emploion head  lmol(⋅) FragEncoder, to compute the representation of each fragment-pair view For the fragment view branch on the right, instead of randomly generating views as other contrastive learning models do, in FraSICL we enumerate and break all of the fragment view Then, to better leverage all of the information related to functional groups contained in the S will be used to compute auxiliary similarity loss, which will be introduced in the next sub-section.In addition, representations NT-Xent loss. The training of FraSICL in the pre-training phase is illustrated in N molecules, the model will calculate projections Similarity loss. In addition, although the representations of different fragment-pair views have been fused, from the perspective of contrastive learning, the representations of different fragment-pair views of the same molecule should also be as close as possible. And as demonstrated in our previous study  molecular representation model is developed. FraSICL is an asymmetric model with two branches, the molecule view branch and the fragment view branch. A multi-view fusion mechanism is also introduced to make better use of the information contained in the views of different fragment pairs. Furthermore, an auxiliary similarity loss is designed to train the model to produce better representations.Baseline experiments are conducted on seven target tasks, and experimental results show that FraSICL achieves state-of-the-art predictive performance with the least amount of pre-training data. Further experiments demonstrate that model finetuning is effective in boosting performance and the auxiliary similarity loss can improve the predictive accuracy if a proper hyperparameter btad462_Supplementary_DataClick here for additional data file.


POLE mutant (POLE), mismatch repair deficient (MMRd), p53 mutant (p53abn), and no specific molecular profile (NSMP). According to the WHO algorithm, 219 ECs were subdivided into the following molecular subgroups: 7.8% POLE, 31% MMRd, 21% p53abn, 40.2% NSMP. Molecular classes as well as ESGO/ESTRO/ESP 2020 risk groups were statistically correlated with disease-free survival. Considering the impact of histopathologic features in each molecular class, stage was found to be the strongest prognostic factor in MMRd ECs, whereas in the p53abn subgroup, only lymph node status was associated with recurrent disease. Interestingly, in the NSMP tumor, several histopathologic features were correlated with recurrence: histotype, grade, stage, tumor necrosis, and substantial lymphovascular space invasion. Considering early-stage NSMP ECs, substantial lymphovascular space invasion was the only independent prognostic factor. Our study supports the prognostic importance of EC molecular classification and demonstrated the essential role of histopathologic assessment in patients’ management.The molecular characterization of endometrial carcinoma (EC) has recently been included in the ESGO/ESTRO/ESP guidelines. The study aims to evaluate the impact of integrated molecular and pathologic risk stratification in the clinical practice and the relevance of pathologic parameters in predicting prognosis in each EC molecular subgroup. ECs were classified using immunohistochemistry and next-generation sequencing into the four molecular classes:  Endometrial carcinoma (EC) represents the most common gynecological cancer in Western countries with an increased incidence in recent years . UsuallyPOLE mutant (POLE) tumors with an excellent prognosis, mismatch repair deficient (MMRd) and no specific molecular profile, (NSMP) tumors with intermediate prognosis, and p53 mutant (p53abn) with poor prognosis [POLE sequencing and immunohistochemistry for p53 and MMR proteins, resulting in a practical and clinically useful molecular classification tool [Recently, new insights into the molecular landscape of endometrial carcinoma are revolutionizing the diagnostic–therapeutic approach to this cancer. The integrated genomic characterization of The Cancer Genome Atlas (TCGA) has defined four distinct prognostic subgroups of endometrial carcinoma: rognosis . Using mion tool ,11,12. ITherefore, the present study aims to evaluate the relevance of molecular classification and traditional pathological factors in a cohort of patients with endometrial carcinoma treated at a referral center. Specifically, the objectives of this study were: (1) to evaluate the role of an integrated risk assessment algorithm in clinical practice that includes both pathologic features and molecular classes; (2) to explore the prognostic impact of pathologic parameters in each EC molecular subgroup to achieve more personalized management of patients.The present study recruited all cases of primary endometrial carcinoma from January 2018 to September 2022. The patient cohort consisted of a preliminary group of retrospectively studied patients  and a suHistologic typing was performed according to the 2020 World Health Organization (WHO) classification of female genital tumors . Tumor gLymphovascular space invasion (LVSI) was defined by the presence of tumor cells within endothelial-lined vascular/lymphatic spaces beyond the tumor invasive front. A 3-tiered system, based on the recommendations by Bosse et al. , was useThe pattern of myometrial invasion was reported, specifying the presence of so-called microcystic, elongated, and fragmented (MELF) glands  and/or aExtensive tumor necrosis has been reported; necrosis only within the glands or on the tumor surface has not been evaluated.The number of mitoses per 10 high-power fields (×400 magnification) served as the mitotic index’s unit of measurement.Four-micron serial sections were cut from each paraffin block and rapidly processed in an automated Benchmark Ultra immunostainer  for immunohistochemical expression of p53, PTEN, MLH1, PMS2, MSH2, and MSH6 using Ventana antibodies and OptiView DAB detection kit (brown color) . Sections were counterstained using Hematoxylin and Bluing reagent following Ventana indications. Immunohistochemical staining of p53 was evaluated as abnormal/mutant-like (p53abn) in case of: (i) protein overexpression, (ii) “null” phenotype, or (iii) positive cytoplasmic staining; otherwise, it was considered normal (wild-type) .PTEN expression was defined: (i) positive (uniform or heterogeneous staining in the neoplastic cells); (ii) negative (no cytoplasmic/nuclear immunostaining) . MismatcBRAF (exon 15), cKIT , CTNNB1 , HRAS (exons 2–4), KRAS (exons 2–4), NRAS (exons 2–4), PIK3CA , POLE (exons 9–14), and TP53 (exons 4–9) are among the genes with the biggest CDS region [http://software.broadinstitute.org/software/igv/ accessed on 1 February 2023) were used to analyze the obtained sequences. The Varsome database , was used to evaluate the pathogenicity of each mutation.From two to four 10-µm-thick FFPE tissue sections were used for DNA extraction, according to the neoplastic areas selected on the hematoxylin and eosin (H&E) control-stained slide. A lab-developed panel including the following genomic areas was used to amplify about 30 ng of gDNA : S region ,23. The S region ,23. OnlyPOLE, MMRd, p53abn, and NSMP subgroups were classified according to the molecular classification included in the 5th edition of the WHO Classification of Female Genital Tumors [POLE mutations to identify the POLE subgroup [POLE mutations, IHC analysis for MMR proteins was used to define MMR deficient (MMRd) tumors. Subsequently, IHC for p53 was performed to detect p53abn carcinomas. Tumors with normal IHC expression of p53 and MMR and no POLE mutations were defined as “no specific molecular profile” (NSMP).l Tumors . As showsubgroup . In the t-test, Mann–Whitney test, and Kruskal–Wallis test were used for comparison between groups. The Kaplan–Meier method with the log-rank test was used to create survival curves; all recurrences  were treated as events. All given p values were based on two-sided tests, and statistical significance was defined as p < 0.05. Analyses were performed using Stata software .Summary statistics are reported as numbers (percentages) or mean ± standard deviation (SD). Chi-squared test, Fisher’s exact test, Clinicopathologic features and molecular classification of the 219 cases of endometrial carcinoma are detailed in 2 = 54.88, p-value < 0.001; see FIGO stage was significantly associated with disease-free survival  MMRd group, 46 (21.0%) p53abn group, 88 (40.2%) NSMP group. Molecular groups are associated with different clinicopathologic parameters  shown in POLE-mutated and p53 abnormal, 2 were triple positive , and one was POLE-mutated and MMRd.Following the 2020 WHO algorithm, EC cases were classified into the following molecular subgroups: 17 (7.8%) Illustrative histopathologic features of EC molecular subgroups are shown in 2 = 26.07, p-value < 0.001; see POLE-mutated cases relapsed, while disease recurrence was identified in 19 (41.3%) p53abn, 10 (11.4%) NSMP, and 9 (13.2%) MMRd cases.Molecular subgroups were significantly correlated with disease-free survival  low-risk, 19 (8.7%) intermediate-risk, 23 (10.5%) high-intermediate, 77 (35.2%) high-risk and 9 (4.1%) advanced/metastatic. ESGO/ESTRO/ESP 2020 risk groups were significantly correlated with disease-free survival .In MMRd subgroup, FIGO stage and lymph node status were correlated with disease recurrence , extensive tumor necrosis  NSMP carcinomas, multivariate analysis showed lymphovascular space invasion results to be an independent prognostic factor see .CTNNB1 exon 3 mutations, L1CAM overexpression, lack of estrogen receptor (ER) and progesterone receptor (PR) expression, ARID1A alterations, chromosome 1q amplification or other copy number alterations, miRNA expression [POLE mutated carcinomas are characterized by lower age at diagnosis, low BMI, and are usually high-grade, endometrioid, and undifferentiated/dedifferentiated histotype. MMRd subgroup has similar features to POLE carcinomas, but patients have older age at diagnosis and higher BMI. MMRd tumors are associated with substantial LVSI and a higher rate of lymph node metastases. p53abn carcinomas arise in older patients with low BMI, tumors are high-grade, histologically serous, endometrioid, and carcinosarcoma, and are characterized by aggressive histopathologic parameters . NSMP tumors represent the majority of cases, related to higher BMI, and they are predominantly early-stage low-grade endometrioid carcinomas. Consistently with literature data, we also demonstrated that molecular subgroups have a significant prognostic impact in our cohort: POLE carcinomas had excellent prognosis without any recurrent event, MMRd and NSMP carcinomas showed intermediate prognosis, and p53abn carcinomas presented worse prognosis. The heterogeneity of the NSMP group comprising both relatively indolent and aggressive ECs also emerges in our cohort. Furthermore, according to the ESGO/ESTRO/ESP guidelines, the combination of clinicopathologic and molecular characteristics has been proven to be essential in accurately assessing prognostic risk classes [POLE carcinomas and did not reach statistical significance in MMRd and p53abn subgroups. Our novel finding that substantial LVSI is an independent adverse prognostic factor in stage I/II NSMP carcinomas implies that this parameter would be reproducible among pathologists, as demonstrated by recent work [The molecular classification of endometrial carcinoma emerged from The Cancer Genome Atlas (TCGA) study provides attractive and useful prognostic insights with relevant diagnostic and therapeutic impact. The molecular subgrouping is considered potentially superior to conventional typing and histologic grading. However, this classifier does not replace clinicopathologic risk assessment based on conventional histopathologic parameters. In this regard, the recent ESGO/ESTRO/ESP guidelines have proposed risk assessment based on the integration of clinicopathologic and molecular features ,25. Furtpression ,29,30,31 classes ,33,34,35 classes . Among d classes ,37,38. O classes ,40. The ent work , and itsIn conclusion, our study showed the importance of applying molecular classification of endometrial carcinoma for precision medicine; however, to date, the accurate and consistent reporting of pathologic parameters, including the evaluation of LVSI, is still vital to ensure optimal patient management.


Dopamine plays a central and multifaceted role in several cognitive functions. It is synthesized and released by dopaminergic neurons that originate from different brain regions, mainly the ventral tegmental area (VTA) and the substantia nigra, and innervate several brain areas, including the nucleus accumbens, amygdala, medial prefrontal cortex, striatum, and hippocampus  have different patterns of D1/5R expression in the hippocampus and found that such altered balance results in changes in dopaminergic-dependent synaptic plasticity. Thus, differences in D1/D5R expression may explain, in part, strain-dependent variations in synaptic plasticity in the murine hippocampus. The complex relationship between reward, motivation, dopamine, and learning prediction error coding remains a challenging question. The contribution by de Oliveira Alves et al. examines the influence of sex and the estrous cycle on the effects of a D2R-antagonist on contextual fear conditioning in rodents and reports that the involvement of D2R during contextual conditioned freezing response shows specificity in female rats. They show that a D2R-antagonist reduces fear expression only in female rats, especially during the proestrus/estrus phases. These articles show that variations in dopaminergic receptor expression across strains, sexes, and individuals have an important— and perhaps underappreciated—role in synaptic plasticity and cognitive functions.How the different dopamine receptors affect brain functions is not well understood and was one of the questions addressed in the current Research Topic. Aberg and Paz report that average reward rates enable motivational transfer across independent reinforcement learning tasks. The mini-review by Zafiri and Duvarci explores the role of various dopaminergic circuits in associative aversive learning, focusing on the amygdala and medial prefrontal cortex circuitry. They highlight how understanding neural circuitry underlying dopamine-dependent aversive learning and memory has high clinical relevance. The review by Sheynikhovich et al. examines dopaminergic modulation of the prefrontal cortex in relation to synaptic plasticities and executive functions, such as spatial strategy selection, memory formation and storage, and extinction learning. The authors show how rodent mPFC is directly involved in long-term memory storage and that memory formation is highly dependent on dopaminergic innervation. In the contribution by Park, the circuit-specific role of D1R on learning and behavioral flexibility was investigated. The author reports that environmental novelty evokes learning-associated plasticity in the vHPC-mPFC and VTA-vHPC circuits, which is dependent on the activation of D1R in the ventral hippocampus. They postulate that this process helps to flexibly overcome previously established spatial bias.Another research challenge comprises understanding how dopaminergic neuromodulation in different brain circuits affects cognition. Aberg and Paz describe how the average reward rate (ARR) affects subsequent motivation and performance in a novel human reinforcement learning paradigm. They found that a low reward probability impairs performance and that this effect can transfer across independent tasks. The study by Isotalus et al. asked the important question of whether L-DOPA, a dopamine precursor used to treat Parkinson's disease, affects sleep and memory. They show that the timing of L-DOPA application relative to sleep augments non-REM sleep and has the potential to disrupt long-term memory in patients with Parkinson's Disease.This Research Topic also explored the role of dopamine in human health and disease. Open questions about the role of dopamine in experience encoding, memory, and cognition can be grouped into two main research domains: fundamental neuroscience and clinical applications. Clinical applications could target three main groups of disorders in the following ways: (1) movement disorders within the nigro-striatal system and dopamine-based therapies require further scrutiny concerning their efficacy in diseases such as Parkinson's disease, dystonia, and Huntington's disease. (2) Memory impairments in neurodegenerative disorders require more clinical trials to elucidate how the pharmacological regulation of dopaminergic neurotransmission may aid patients suffering from neurodegenerative dementias. (3) Psychiatric disorders such as depression, anxiety, panic disorder, and PTSD would strongly benefit from more clinical trials exploring the dopaminergic control of memory consolidation for aversive memories. In fundamental neuroscience, dopamine could be subjected to further scrutiny at three levels: (1) on a cellular level, one should further address how D1/5R and D2R affect different types of neurons and information processing; (2) on a network level, one should further explore how dopamine regulates the functional connectivity of large neuronal populations, particularly in structures that are known to mediate behavior, such as the extended hippocampal system, prefrontal cortex, amygdala, and striatum; (3) on a behavioral level, how theoretical models of reinforcement learning relate to their biological substrates, and how dopamine shapes these to modulate cognitive function, should be addressed in more detail.In conclusion, this collection of articles highlights the crucial role of dopaminergic modulation in different aspects of synaptic plasticity and cognition. The studies reveal that variations in the expression of dopaminergic receptor subtypes across rodent strains, sexes, and individuals can have profound effects on brain function related to learning and memory. The articles also shed light on the role of dopamine in human motivation, sleep, and brain dysfunction, pointing toward potential therapeutic targets. Overall, these findings open up exciting avenues for future research in the field of dopamine and cognitive neuroscience.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.