Ligand-based virtual screening (LBVS) is a promising approach for rapid and low-cost screening of potentially bioactive molecules in the early stage of drug discovery. Compared with traditional similarity-based machine learning methods, deep learning frameworks for LBVS can more effectively extract high-order molecule structure representations from molecular fingerprints or structures. However, the 3D conformation of a molecule largely influences its bioactivity and physical properties, and has rarely been considered in previous deep learning-based LBVS methods. Moreover, the relative bioactivity benchmark dataset is still lacking. To address these issues, we introduce a novel end-to-end deep learning architecture trained from molecular conformers for LBVS. We first extracted molecule conformers from multiple public molecular bioactivity data and consolidated them into a large-scale bioactivity benchmark dataset, which totally includes millions of endpoints and molecules corresponding to 954 targets. Then, we devised a deep learning-based LBVS called EquiVS to learn molecule representations from conformers for bioactivity prediction. Specifically, graph convolutional network (GCN) and equivariant graph neural network (EGNN) are sequentially stacked to learn high-order molecule-level and conformer-level representations, followed with attention-based deep multiple-instance learning (MIL) to aggregate these representations and then predict the potential bioactivity for the query molecule on a given target. We conducted various experiments to validate the data quality of our benchmark dataset, and confirmed EquiVS achieved better performance compared with 10 traditional machine learning or deep learning-based LBVS methods. Further ablation studies demonstrate the significant contribution of molecular conformation for bioactivity prediction, as well as the reasonability and non-redundancy of deep learning architecture in EquiVS. Finally, a model interpretation case study on CDK2 shows the potential of EquiVS in optimal conformer discovery. The overall study shows that our proposed benchmark dataset and EquiVS method have promising prospects in virtual screening applications.  Virtual screening  adopts cWith the massive development of public pharmaceutical databases and artificial intelligence technologies, data-driven deep learning frameworks have been widely used in the vast majority fields of drug discovery, such as de novo drug design ,12, ADMEbioactivity prediction benchmark dataset with multiple molecular conformers is still lacking, and the end-to-end deep learning architecture that effectively learns molecular representations from molecular conformers has not been designed and estimated.However, the molecular 3D structures-molecular conformers, have seldom been considered in these studies. Previous studies have proven that there were certain relations between different molecular conformers and bioactivities ,27,28, aTo address the above challenges, we first constructed a large-scale molecular bioactivity benchmark dataset with over 3 million endpoints, including nearly 1 thousand targets, 1 million molecules, and 10 million calculated molecular conformers. Then, we proposed a new deep learning method called EquiVS to introduce molecular conformation information into LBVS, which effectively improved molecular bioactivity prediction. EquiVS was designed as an end-to-end architecture with graph convolutional network (GCN), equivariant graph neural network (EGNN), and deep multiple instance learning (MIL) layers, which can directly learn molecular high-order representations from the 2D topological level and 3D structural level. The model performance comparison results on large-scale benchmark datasets indicated that EquiVS outperformed multiple classical machine learning-based and graph neural network-based baseline methods. Furthermore, the ablation study emphasized that efficient representations and attention-based aggregations of multiple molecular conformers play an important role in the accurate bioactivity prediction in EquiVS. To enhance the interpretability of EquiVS in real LBVS scenarios, we introduced an attention-based mechanism in deep MIL for optimal molecular conformer discovery which was investigated by a case study.We examined the quality of our constructed bioactivity benchmark dataset through visualization analysis. Extensive chemical structure  and bioactivity distribution  analysis for the initial source bioactivity data can be found in . In thisMeanwhile, As for the selection of candidate targets, Hence, eight physiochemical and structural properties were calculated for all molecules in the benchmark dataset to explore the distributions of Linpski rules  and QuanWe then trained EquiVS and 10 baseline models on our organized bioactivity benchmark dataset to compare the overall performances of these LBVS bioactivity prediction models, which were listed in 2: GBDT_ECFP, MSE: GBDT_ECFP, MAE: GAT), the relative improvements of R2, MSE, and MAE of EquiVS reached 3.60%, 33.01%, and 5.45%, respectively. Also, the performances of EquiVS are generally stabler than other deep learning methods.Furthermore, as the qualities and confidences of bioactivity endpoints varied in different bioactivity sub-dataset, there are a proportion of bioactivity sub-datasets that failed to be used to train bioactivity prediction models with sufficient performances. Therefore, it is necessary to exclude low-quality bioactivity sub-datasets corresponding to infeasible targets and focus on those feasible ones which have sufficient bioactivity data with fewer noises and better consistencies. Considering this, we set The overall model performance comparison experiments emphasized that our designed EquiVS method can give relatively accurate predictions on molecular bioactivities rather than widely used machine learning and deep learning-based baseline methods. Especially in the scenario of bioactivity prediction with good-quality training data, the superiority of EquiVS is highlighted.EquiVS-Single: EquiVS architecture which adopts one conformer for each molecule for structural representation learning.EquiVS-w/o GCN: EquiVS architecture which replaces the GCN layer with a simple linear layer.EquiVS-w/o Skip: EquiVS architecture which deletes the skip connection between different EGNN layers.EquiVS-w/o AA: EquiVS architecture which replaces the attention-based conformer representation aggregation process with a simple sum calculation.EquiVS-w/o IP: EquiVS architecture which deletes the conformer-level predictor and the corresponding conformer-level prediction loss.After the effectiveness of EquiVS on large-scale bioactivity prediction has been proven, we then explored the potential mechanisms and core modules that play important roles in the molecular representation learning and model performances of EquiVS through an ablation study. Specifically, we designed five variant models, and each of them deleted or replaced a core module in EquiVS and left other modules. These variant models are as follows:After these variants were defined, considering the running time and computing complexity, we randomly selected 50 targets and their bioactivity sub-datasets and trained EquiVS and these variants on them for performance comparisons. The data splitting settings are the same as those in model performance comparison experiments. The performances of EquiVS and its 5 variants were shown in 2, 35.14% on MSE, and 27.94% on MAE. The results indicated that multiple sampling of molecular conformers can enhance molecular representation learning, and it is easier than a single conformer to obtain the right conformer which matches the bioactivity endpoint best, thus “diluting” the potential molecular three-dimensional structural noises caused by unsupervised conformer generation methods, which is also supported by a previous study  n2)cc1” as a case active molecule , visualized its conformers, predicted the bioactivity values by the conformer-level predictor in EquiVS, generated the binding score and pose by Autodock Vina , to inveThe overall results suggested that the attention-based conformer-level model interpretability of EquiVS can differentiate between molecular conformers with varied structure reliabilities and discover the optimal conformers for identifying specific bioactivity endpoints and targets. Compared to the machine learning-based LBVS methods, which could not directly handle molecular 3D structures and lacked interpretabilities, the attention mechanism in EquiVS provides valuable insights regarding the optimal conformers and discovery of binding poses.We introduced the data collection, integration, filtering, and preprocessing process of the molecular bioactivity benchmark dataset for drug virtual screening in detail. In the first steps, we collected large-scale molecular bioactivity data from five public databases, including ChEMBL , PubChemAs for the data integration, in , the bioIn the second step, we proposed a multilevel data filtering strategy to identify high-quality bioactivity data. For bioactivity data, we selected endpoints with five widely used and studied activity types  as candidates and filtered the last ones. For molecular structure, we adopted MolVS  for struCandidate bioactivity endpoints were determined by the above comprehensive process. However, these data contained only one-dimensional SMILES representations of molecular structures and lacked three-dimensional structural information. To address this issue, multiple molecular conformers were computationally generated and assembled with bioactivity endpoints to finish the bioactivity benchmark dataset construction. In this study, we used the RDKit package to generate and optimize molecular conformers. Specifically, hydrogen atoms were first added to the molecular structure to simulate the real molecular geometric conformation. Then, ETKDG  was used for conformation generation. ETKDG is a knowledge-based conformer generation method that combines distance geometry and torsion angle preferences proposed by Riniker et al . AdditioHere, we aligned the three-dimensional coordinates of different conformers of each molecule to normalize coordinate information. All hydrogen atoms were removed from the molecular structures to reduce the computing complexity and training time of LBVS methods. Finally, we assembled all conformers to corresponding molecule SDF structure files to finish the construction of the large-scale bioactivity benchmark dataset.To introduce the molecular conformer information to our GNN-based EquiVS method, we constructed each molecular graph with an adjacency matrix, a node feature matrix Given a molecule, its graph As for feature matrices in the molecular graph, we first calculated atom physiochemical properties as In this study, we proposed an EGNN and deep MIL-based virtual screening method, EquiVS, to facilitate the representation learning of current LBVS methods via utilizing molecular conformations. The architecture of EquiVS  compriseGraph representation learning module. We considered learning molecule high-order representations from both two-dimensional topological level and three-dimensional structural level. Therefore, a stepwise molecular graph learning strategy with skip connection was designed to learn and aggregate the molecular representations. Specifically, a GCN layer is first used to learn graph topological representations Finally, as the “over-smoothing” and “vanishing gradient” widely exist in deep graph neural network models, which could significantly harm the model performances, we built skip connections between different EGNN layers to allow direct gradient propagation to the shallow layers by simply concatenating the graph feature output of each layer as the final graph representations. For conformer Deep multiple instance learning module. After EquiVS captures the high-order representations for molecular conformers, the aggregation process of these conformer representations to generate comprehensive molecular representations, and effective bioactivity prediction through both conformer-level and molecule-level should be elaborately considered and designed. Regarding this, we introduced MIL theory into bioactivity prediction and designed our deep multiple instance learning module with an interpretable attention mechanism. Based on MIL theory, a molecule can be regarded as a “bag”, and its multiple conformers are “instances”. The bioactivity value is only labeled at the molecule level, but the conformer-level bioactivities still remain unknown. Therefore, the training object of MIL is to accurately predict the molecular bioactivity value, while identifying the conformer which fits this bioactivity value best, simultaneously. Regarding this, EquiVS first dynamically aggregates the conformer instance representations with an attention mechanism. The attention score of The attention coefficient represents the importance weight of a conformer. Hence, the conformer instance representations can be aggregated to acquire molecule representation Then, EquiVS predicts the bioactivity value from both conformer-level and molecule-level. For conformer-level prediction, a multilayer perceptron (MLP) is used for bioactivity prediction:Inspired by loss-based deep multiple instance learning , we adopAlgorithm 1: The computing flow of EquiVSInput: A molecule graph Output: Predicted bioactivity   2: Acquire 2D topological graph representation Equation (3).   3: for Molecular conformers do  4:    for EGNN layerdo  5:        Acquire 3D structural graph representation       Equation (8).   6:   end for  7:   Concatenate the conformer representation Equation (13).   8: end for  9: Acuiqre aggregated graph representation with attention mechanism     Equations (14)–(16).10: Generate conformer-level prediction Equation (17).11: Generate molecule-level prediction Equation (18).12: Calculate conformer-lebel loss Equations (19) and (20).13: Update parameters by optimizing Equation (21).14: Ouput   1: Initialize the trainable parameters in EquiVS. The overall EquiVS computing flow was shown in Algorithm 1. Furthermore, we used Adam optimizer for training optimization and added a dropout layer after each GNN layer and MLP layer to alleviate overfitting.Deep multiple instance learning module, EquiVS employs the attention mechanism to aggregate the conformer representations, which could also be used for conformer-level model interpretation. Given a bioactivity endpoint with a molecule and multiple conformers, we ranked the importance of different conformers based on the attention coefficients As described in For model hyper-parameter settings, we set the learning rate as 0.001, the dropout rate as 0.05, the number of epochs as 200, batch size as 128, the hidden feature dimensions as 128, and the number of EGNN layers as 2 to construct and train EquiVS on our proposed bioactivity prediction benchmark dataset.For baseline settings, we adopted two molecular fingerprints (ECFP4  and MACC2), MSE, and mean absolute error (MAE) to assess the performances of bioactivity prediction. These metrics can be formulated as:For experimental settings, we randomly split the benchmark dataset into training set, validating set, and testing set with a ratio of 8:1:1. EquiVS and all baseline methods were trained on the training set, adjusting hyper-parameters on the validating set, and evaluated on the testing set. We adopted the coefficient of determination (RIn this study, we investigate the role of molecular conformation in LBVS and bioactivity prediction scenarios. A large-scale bioactivity prediction benchmark dataset is proposed to assemble the requirement of molecular conformation to bioactivity endpoints, which is collected from multiple public pharmaceutical databases and contains thousands of targets and millions of bioactivity endpoints, molecules, and molecule conformers. Then, an EGNN and deep MIL-based LBVS method is designed for bioactivity prediction, which is called EquiVS. Compared to other widely-used ML-based and GNN-based methods, EquiVS achieved notable improvements on our large-scale benchmark dataset. Combining the ablation analysis, the performance results prove employing molecular conformation could enhance molecular representation learning and further contribute to better bioactivity prediction with elaborate neural network architecture design and reasonable feature extraction and aggregation. To promote the practical application of EquiVS, two case studies are designed to explore the effectiveness of conformer-level interpretation in EquiVS. The overall results reveal a promising prospect of molecular conformation as well as our proposed benchmark dataset and EquiVS method in bioactivity prediction and LBVS.It should also be emphasized that there are several major limitations in our study. First, from the data quality control aspect, some of the sub-datasets and targets in our integrated benchmark dataset should be filtered as all tested LBVS methods could not give reliable predictions on them, but they are still retained in the current version; Second, from the model training aspect, a predictable optimization is to pre-train EGNN-based models using molecular conformation and finetune them for the downstream bioactivity prediction ,58,59. AAs for future research, we will focus on developing practical tools to enhance the accessibility of EquiVS and employing molecular conformation using massive unlabeled molecules to pre-train and optimize EquiVS models in terms of robustness and generality.


Molecular Oncology, a journal wholly owned by the Federation of European Biochemical Societies (FEBS).The formation of organisations and societies within all areas of scientific research facilitates the bringing together of researchers in a given field and serves to aid communication, collaboration, progress of science and career development. Even greater gain can be attained when individual organisations form partnerships to complement each other's activities and to increase the scope of their endeavours. Within this editorial, we highlight the key points of a new partnership formed between two non‐profit bodies within cancer research, the European Association for Cancer Research (EACR) and  Molecular Oncology, a journal wholly owned by the Federation of European Biochemical Societies (FEBS).The formation of organisations and societies facilitates bringing researchers together and aids communication, collaboration, scientific progress and career development. In this editorial, we highlight key points of a newly formed partnership between two charitable organisations within cancer research, the European Association for Cancer Research (EACR) and  The European Association for Cancer Research's mission focusses on ‘The advancement of cancer research for the public benefit: from basic research to prevention, treatment and care’. The organisation has gone from strength to strength since its inception and now has over 12 000 members worldwide. It continues to promote in‐person and virtual collaboration and networking between cancer researchers, as well as offering a variety of funding opportunities to support its members. For example, the EACR supports travel fellowships and travel awards that assist early career researchers in gaining greater experience by visits to other laboratories or by attending scientific events. The EACR itself also organises a number of scientific events throughout the year, with the annual EACR Congress being the association's landmark event. The 2023 EACR Congress is the 29th annual congress and will take place in Torino, Italy, between the 12th and 15th of June.The European Association for Cancer Research (EACR) was founded in 1968 to enable a greater collaboration between cancer researchers across geographical Europe (https://www.febs.org/). The Federation of European Biochemical Societies is made up of 39 Constituent Societies across Europe and nearby states, which culminates in a membership of more than 35 000 researchers. The Federation of European Biochemical Societies is supported by FEBS Press, which publishes four journals: FEBS Journal, FEBS Letters, FEBS Open Bio and Molecular Oncology. Molecular Oncology is a multidisciplinary cancer journal publishing basic, translational and clinical cancer research as well as policy articles reporting developments in the cancer research community. All profits from the four journals are reinvested back into science to support fellowships, travel grants, FEBS Advanced Courses, the annual FEBS Congress (https://2023.febscongress.org/) and other FEBS Initiatives.Similar to the EACR, the Federation of European Biochemical Societies (FEBS), founded in 1964, is a non‐profit society focussed on biochemistry and molecular biology . Due to this session's great success and since the EACR and FEBS have similar goals in supporting science, further discussions ensued to see whether there could be additional common goals between the EACR and Molecular Oncology that they could work together towards. This culminated in a memorandum of understanding between Molecular Oncology, FEBS and the EACR covering various points that led to the recognition of Molecular Oncology as an ‘Affiliate Journal’ of the EACR.Initial interactions between Molecular Oncology/EACR/EACS session at the 2022 EACR Congress, it was agreed to hold a second joint session at the 2023 Congress on the topic of ‘Challenges of Data Management and Sharing for Translational Cancer Research’ [Molecular Oncology editorial office will also be in attendance at the Congress, and Kevin Ryan, Editor‐in‐Chief of Molecular Oncology, will take part in a ‘Meet the Editors’ panel discussion together with editors and publishers from other cancer journals . Molecular Oncology will also publish the abstract proceedings of the EACR Congress as a supplement in a special issue.Following the success of the joint esearch’ . MembersMolecular Oncology, the journal has recently started to publish a new article type entitled ‘Viewpoints from the EACR’, in which researchers connected to the EACR are invited to write commentaries on recent developments or contentious topics in cancer research and their transformative opinions. Two of these have already been published [In further connections between the EACR and ublished , 3, and Molecular Oncology will also form an Editorial Academy inviting EACR early career members to gain experience in publishing and journal editing. Moreover, the Molecular Oncology editorial office will also organise a small number of publishing internships to further support this goal by hosting EACR members as interns at the journal to familiarise themselves with the other side of peer review. Further collaborations between the EACR Early Career Researchers' Council and the Molecular Oncology Early Career Advisory Board will also be encouraged and developed.In recognition of the fact that both FEBS and the EACR have a central focus on supporting early career researchers, The aim of this partnership between the EACR and Molecular Oncology is to strengthen collaboration and facilitate progress in cancer research within Europe and beyond. Both parties have agreed to promote each other's individual, as well as joint initiatives, and all involved hope that this collaboration grows and brings great positivity in the years to come.The authors declare no conflict of interest.


Introduction: Opioids are the leading cause of overdose death in the United States, accounting for almost 70,000 deaths in 2020. Deep brain stimulation (DBS) is a promising new treatment for substance use disorders. Here, we hypothesized that VTA DBS would modulate both the dopaminergic and respiratory effect of oxycodone.Methods: Multiple-cyclic square wave voltammetry (M-CSWV) was used to investigate how deep brain stimulation  of the rodent ventral segmental area (VTA), which contains abundant dopaminergic neurons, modulates the acute effects of oxycodone administration  on nucleus accumbens core (NAcc) tonic extracellular dopamine levels and respiratory rate in urethane-anesthetized rats .Results: I.V. administration of oxycodone resulted in an increase in NAcc tonic dopamine levels (296.9 ± 37.0 nM) compared to baseline (150.7 ± 15.5 nM) and saline administration (152.0 ± 16.1 nM) . This robust oxycodone-induced increase in NAcc dopamine concentration was associated with a sharp reduction in respiratory rate . Continuous DBS targeted at the VTA (n = 5) reduced baseline dopamine levels, attenuated the oxycodone-induced increase in dopamine levels to (+39.0% vs. +95%), and respiratory depression .Discussion: Here we demonstrated VTA DBS alleviates oxycodone-induced increases in NAcc dopamine levels and reverses respiratory suppression. These results support the possibility of using neuromodulation technology for treatment of drug addiction. Opioids are the leading cause of overdose death in the United States, accounting for almost 70,000 deaths in 2020 . The maiHowever, electrical stimulation is inherently non-selective in what neuronal elements and terminals are activated. Thus, Nac DBS is releasing many different neurotransmitters in the Nac  which does not address what neurotransmitter may be the most relevant in mitigating addiction. In opioid use disorder, dopamine is one of the key monoamine neurotransmitters that mediates incentive-motivation which plays an important role in drug reinforcement and compulsive drug-seeking behaviors. Opioids are known to stimulate µ-opioid receptors on GABAergic nerve terminals synapsing on dopamine neuronal cells in the ventral tegmental area (VTA) producing both dopamine-dependent and dopamine-independent positive reinforcement . Activat2 and pH dependent respiratory drive  and respiratory rate changes caused by acute intravenous oxycodone administration.In the current proof-of-concept study, we hypothesized that VTA DBS would provide modulation of the oxycodone effect on tonic dopamine levels, as well as its ability to reduce the suppressive effect on respiration. To achieve this, we first elucidated oxycodone-induced dopaminergic changes n = 10, weighing 250–300 g; Envigo, IN, United States) were used for this study. Rats were kept in group housing in an Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC) accredited vivarium following a standard 12-h light/dark cycle at constant temperature (21°C) and humidity (45%) with ad libitum food and water. Animal studies were approved by the Institutional Animal Care and Use Committee (IACUC), Mayo Clinic, Rochester. The NIH Guide for the Care and Use of Laboratory Animals guidelines  were followed for all aspects of animal care.Adult male Sprague-Dawley rats  and administered buprenorphine  for analgesia. Following anesthesia, the rat was placed in a stereotaxic frame . Respiratory rate  and hind-paw and tail pinch were used to monitor the physiological state and depth of anesthesia, respectively. RespiRAT is a pressure monitor that only counts the peaks of pressure to mitigate interference from noise, and automatically adjusts the detection threshold with the amplitude of the incoming data. Using a standard rat brain atlas , three tThe depths of the stimulating electrode in the VTA and CFM in the NAcc were first adjusted to obtain a robust stimulation-evoked dopamine signal as measured by fast scan cyclic voltammetry (FSCV) . Stimulation parameters were biphasic pulses at 60 Hz, 2 ms pulse width, 0.2 mA, and 2 s duration. Stimulation and FSCV were both performed using the WINCS Harmoni system , a wirelOnce the optimal electrode depths were identified, the system was switched to the M-CSWV sensing technique . After aExperiment 2 consisted of the stimulation group , VTA biphasic pulse stimulation was applied at 130 Hz  continuously for 60 min. The delivered stimulation was interleaved with the M-CSWV recording to minimize artifacts. Once the signal was restabilized to a new plateau (after 30 min of stimulation), i.v. oxycodone was administered (dose and method as above) while stimulation and recording continued. After another 30 min of observation, the stimulation was turned off. The animal was observed for another 30 min post-stimulation before being euthanized using Fatalplus injection .Saline administration during VTA stimulation was shown to have no effect on tonic dopamine levels in our previous study and was therefore not repeated here .in vitro with dopamine solutions of different known concentrations. This was performed in a similar manner to previously described procedures .For comparison between different dopamine concentrations, repeated-measures one-way (Experiment 1) and two-way (Experiment 2) ANOVA tests were performed. The levels for comparison were all calculated by averaging over 10 data points, i.e., 100 s. In Experiment 1, the levels include: “Baseline” , “Saline” , and “Oxycodone (Peak)” (centered at the peak of dopamine levels after oxycodone was administered). Post-hoc two-tailed paired In Experiment 2, two-way ANOVA test was performed across: “Baseline” (centered at 5 min prior to start of stimulation), “Stimulation” (centered at 5 min prior to administering oxycodone); “Oxycodone” (centered at the peak of dopamine levels after oxycodone was administered), and “post-stimulation” (centered at 15 min after stimulation stops).t-tests were also conducted. Respiratory rates are measured by calculating the moving average over 30 s. Measurements were taken approximately 1/54 s apart. The levels for comparison were calculated by averaging over 10 data points. In Experiment 1, a comparison was made between the levels post-saline  and the levels post-oxycodone (centered at troughs). Likewise, in Experiment 2, a comparison was made between the levels post-stimulation (10 min prior to oxycodone administration) and the levels post-oxycodone (centered at troughs).For comparison between respiratory rates, two-tailed paired p < 0.05. No Bonferroni corrections to t-tests were applied as sequential comparisons were set up for testing a pre-planned hypothesis .Statistical analyses were performed and graphs were constructed using PRISM 8 (GraphPad). All error bars and shaded areas are represented as S.E.M. Statistical significance was set at p = 0.0224). Post-hoc analysis showed saline did not significantly alter tonic dopamine levels  . This is also highlighted in the high dimensional color plots, and their voltammograms in To determine the viability of VTA DBS as a potential therapy for opioid intoxication, dopamine levels were first monitored in response to oxycodone injection. A saline injection was performed before the oxycodone injection as shown in  0.1047) . In contF = 30.48; p = 0.0053] and oxycodone  both had a statistically significant effect on tonic dopamine levels  = 130.5; p = 0.0003]. This was also reflected in the less intense red color on the color plots . VTA stimulation partially alleviated respiratory suppression from 36.6% to 13.4% relative to baseline (p = 0.072) (p = 0.148).One consideration was that VTA DBS may alleviate the respiration-suppressing effects of acute opioid administration. Respiration rate was monitored and compared for saline and oxycodone treatment in the absence of VTA DBS, and oxycodone treatment during VTA DBS . Oxycodo= 0.072) . The cleThe present study measured tonic dopamine levels in the NAcc and accompanying respiratory rate following i.v. oxycodone administration. The facilitatory increase in dopamine levels is consistent with previous microdialysis studies with opioids . The risphasic (burst) dopamine release  similar to chronic antipsychotic treatments that, in turn, decrease tonic DA levels in the NAcc . These hVTA DBS is currently being used as an experimental treatment for treatment-refractory cluster headache . InteresWith regards to the potential applicability of DBS for substance use disorder, no human studies have been reported using VTA DBS. In preclinical studies, one study demonstrated continuous optogenetic stimulation of VTA dopaminergic neurons can reduce ethanol self-administration in mice . Althougcore was chosen as the main target for dopamine measurements for two main reasons. First, from microdialysis studies, extracellular dopamine in the core was higher than that in the shell, whereas norepinephrine levels  were higher in the shell (In the current study, the NAc he shell . Second,he shell . Althoughe shell .Consistent with previous microdialysis studies , oxycodoThe cases of opioid overdose in the United States have more than quadrupled between 2000 and 2020 and, without further intervention, is expected to double by 2029 . TherefoLimited historical precedence or evidence implicates DBS as a potential procedure for respiration modulation. One study showed subthalamic nucleus stimulation may improve upper respiratory and laryngeal control in Parkinson’s patients , but no This proof-of-concept preclinical study utilized anesthetized, naïve rats as subjects rather than a chronic addiction model. Although a relatively high acute dose of oxycodone was administered that induced a clinically corollary of respiratory depression, there were no measures for addictive behaviors such as drug-seeking and compulsion. Future studies in awake animals, particularly in drug self-administration paradigms, will be required to validate the dopaminergic effects, as well as behavioral and respiratory benefits.While dopamine plays a key role in the process of drug addiction, there are also studies which showed opioids may have a dopamine-independent reward mechanism . TherefoThe present study demonstrated that M-CSWV served as a reliable tool to measure changes in tonic extracellular dopamine levels in the NAcc following acute i.v. oxycodone administration. The observation that VTA DBS at least partially reversed the opioid-induced increase in NAcc dopamine levels and respiratory rate depression provides the basis for exploring the use of DBS to treat opioid use disorder and overdose.


This study numerically demonstrates the light absorption spectra of each base of DNA-wrapped single-walled carbon nanotubes (SWCNTs). Previous experimental and theoretical studies show that the optical properties of these composites are different from the bare SWCNTs. In this work, we investigated the bases of DNA that influence optical properties. To obtain stable molecular states for studying optical properties, molecular dynamics calculations were performed. Additionally, light absorption spectra in the ultraviolet-to-near-infrared region of one type of base-wrapped  SWCNTs were investigated by utilizing the semi-empirical molecular orbital theory using SCIGRESS commercial software. This method can significantly reduce the calculation time compared to the ab initio molecular orbital method, making the handling of composites of bases and SWCNTs possible. We found that the largest peaks appear at a wavelength of around 300 nm for all the composites. Furthermore, we found that the light absorption spectra above 570 nm are strongly influenced by adenine and cytosine. Thus, our computational results provide insight into the optical properties and the effects of base–SWCNTs that are difficult to investigate experimentally under the influence of solvents and various molecules. Single-walled carbon nanotubes (SWCNTs) have potential applications in various industries, such as electronics, nano mechanics, health care, etc. SWCNTs are also useful nanomaterials for biotechnology, aerogels, films, fibers, and energy supply systems ,2,3. SWCOn the other hand, from a macro perspective, our previous research focused on the engineering application of DNA–SWCNT as a biomaterial and analyzed the mechanical vibration properties using the finite element method (FEM) ,9. The iUsing molecular dynamics methods, this study used SCIGRESS in commercial software  to calculate the stable state of SWCNTs wrapped with adenine, thymine, cytosine, or guanine. Further, we calculated the light absorption spectra of these composites over a wide range from ultraviolet to near-infrared light by the semi-empirical molecular orbital theory method. The light absorption spectra calculated by the theory provide the optical properties of each base and SWCNTs under vacuum, omitting the complicated conditions of the experimental solution. We also investigated the effect of each number of bases placed around SWCNTs on their optical absorption spectra. These results are expected to contribute to a fundamental understanding of the optical properties of DNA–SWCNTs composites.Molecular arrangement for six base numbers of all targeted adenine-, thymine-, cytosine-, or guanine-wrapped SWCNTs composites are prepared by SCIGRESS. It is important to note that the molecular arrangement for each base number is determined by the molecular dynamics method. First, we compared the light absorption spectra of three types of bare SWCNTs and base and SWCNTs wrapped by the base. The wavelength peaks of SWCNTs calculated differ by about 15% compared to the experimental results. We believe that the differences are due to experimental and computational conditions variations. All light absorption spectra in our studies were performed in a vacuum, but the experiments were performed in water. This difference implies that the dielectric background of water affects the exciton optical transitions but not the electric dipole–vibronic transitions. Therefore, the effect of the solvent is significant. Moreover, previous studies report that the pH and ionic strength of water also change the light absorption spectra , which mThe present study numerically calculated the wavelength dependence of light of the molar absorbance coefficient of the adenine-, thymine-, cytosine-, or guanine-wrapped SWCNTs. The most crucial finding is that the largest peaks appear at a wavelength of around 300 nm for the case of all composites. This also proves that the numerical simulation is in good agreement with the experimental data, and the peak value of the wavelength does not shift due to the interaction between bases and SWCNTs. The molar absorbance coefficients of adenine and cytosine, indicated by the black lines in An attempt has been made to understand why thymine wrapping has a much weaker effect on the light absorption spectra than other base wrapping. The difference in structure with other bases is affected as the spectra do not change when the thymine is rearranged with respect to SWCNTs. For example, as cytosine and thymine are considered, they have the same six-membered ring structure. However, the difference is that thymine has more oxygen attached, so thymine has fewer electrons taken away, resulting in no change in the spectra. Compared to guanine, which has the next smallest spectral change after thymine, guanine has the same six-membered ring structure as thymine in the way that the addition of a five-membered ring takes up electrons. Although the results presented in this study cannot give a clear interpretation of the effect of the structure of each base on SWCNTs, we could calculate the effect of the interaction of each base with SWCNTs on the spectra. We believe that this contributes to the understanding of future experimental results. These findings are expected to accelerate the study of optical characteristics of base-wrapped DNA–SWCNTs. Furthermore, the previous experimental study showed that the peak of the molar absorbance coefficient was observed near 990 nm . In the Finally, we present future work to further develop this study. The semi-empirical molecular orbital theory method used in this study is a practical method that significantly reduces the computation time compared to the first principles molecular orbital method. However, obtaining the optical absorption spectra of bases and SWCNTs up to near-infrared light requires much calculation time. To solve these problems, material informatics using deep learning may be useful. In research on the light absorption spectra of DNA–SWCNT, a method of creating a Bayesian regularized back-propagation neural network model from a small number of experimental results and predicting various conditions in a data-driven manner has been reported . Based oIn this work, we fix it at 1 nm and show only calculations limited to  SWCNTs. Countless combinations of chirality and lengths are associated with SWCNTs, and it is difficult to calculate all combinations by the molecular orbital method. In addition, only bases were wrapped in SWCNTs. However, in the future, it will be necessary to investigate interactions with other molecules, such as deoxyribonucleic acid and phosphoric acid, and the effects of the state in which multiple bases are linked. Furthermore, there is no doubt that the influence of water molecules around those DNA–SWCNTs is also a crucial factor to investigate. Reproducing these effects with a molecular model will result in a huge molecular weight, and the computation time is predicted to increase significantly. It is likely that the number of cases that can actually be calculated is limited. In this context, machine learning may be required to predict various conditions from a small amount of data. Therefore, future work is expected to utilize machine learning, such as the Bayesian regularized back-propagation neural network model.0 and R0 are fitting parameters given by D0 =0.2483 kcal/mol and R0 = 3.848 Å, respectively. This potential function is widely used since it can express intermolecular interactions simply and accurately. The states with and without consideration of the Lennard–Jones potential are shown in The light absorption spectra of bases of DNA and SWCNTs presented in this study were performed using the molecular dynamics and molecular orbital algorithms provided in SCIGRESS basic (Fujitsu Co. Ltd.). This software is developed based on widely used molecular orbital theory ,20 and nn is the total number of atomic orbitals.The light absorption spectra were calculated using the molecular orbital method for the molecular model optimized by the Lennard–Jones potential. According to standard molecular orbital theory, an orbital of electrons There are three types of molecular orbital methods: ab initio molecular orbital method, semi-empirical molecular orbital, and empirical molecular orbital. In this work, we used the semi-empirical molecular orbital theory, the Hartree–Fock approximation, to find al (STO) . This meIn the semi-empirical method, the zero differential overlap (ZDO) approximation is the usual approach, and the overlap integral matrix becomes just the identity matrix. The Hartree–Fock–Roothaan (HFR) equation is simplified as shown below.C and the orbital energy E. As a computational procedure, first, the initial value of C was computed using a simpler method such as Huckel’s method. This initial guess gave F. Then F was diagonalized to compute C and E. This new C matrix was used to compute F for the next computation of C and E. Finally, an iterative calculation called self-consistent field (SCF) was performed until C converged to satisfy the threshold. By solving this HFR equation, the total electron energy E could be obtained. E was given by the following formula:nbasis is the number of basis functions. This version of the equation forms the starting point for the semi-empirical method. Solving this equation gives the molecular orbital coefficient


The use of molecular testing to direct diagnosis and treatment options in ovarian tumours has rapidly expanded in recent years, in particular with regard to the recommendation for routine homologous recombination deficiency (HRD) testing in all patients with high-grade ovarian epithelial tumours. The implications of this increased level of testing upon the pathologist is significant in terms of increased workload, the provision of adequate tumour samples for molecular testing, and the interpretation of complex molecular pathology reports. In order to optimise the quality of reports generated, it is important to establish clear pathways of communication on both a local and national level between clinicians, pathology lab staff, and medical scientists. On a national level, in the United Kingdom, Genomic Laboratory Hubs (GLHs) have been established to provide a uniform high-quality molecular diagnostics service to all patients with ovarian tumours within the National Health services in the country. On a local level, there are a number of small steps that can be taken to improve the quality of tissues available for testing and to streamline the processes involved in generating requests for molecular testing. This article discusses these factors from the perspective of the clinical histopathologist. According to the Global Cancer Statistics 2020, ovarian cancers are the eight most-common malignancy in women, accounting for 3.4% of all new cancer diagnoses in women worldwide . OvarianThe standard treatment for ovarian cancer is cytoreductive surgery followed by platinum-based chemotherapy; however, due in part to the high intratumoural heterogeneity of these neoplasms, most patients will eventually develop resistance to platinum-based therapies . In lighAll patients with a new diagnosis of high-grade ovarian epithelial cancer  are consented for homologous recombination deficiency (HRD) testing. Homologous recombination is a pathway involved in repairing double-strand DNA breaks, and therefore, its deficiency results in genomic instability, which is a target for PARP inhibitors . HRD canMismatch repair (MMR) deficiency testing is performed routinely in ovarian endometrioid and clear cell carcinomas, both of which are commonly associated with endometriosis ,9. The tOver 97% of HGSCs harbour a TP53 mutation, which is assessed at diagnosis using routine immunohistochemistry . MutatioMucinous ovarian carcinomas are associated with the copy number loss of CDKN2A and KRAS mutations, both of which are believed to be early events in tumorigenesis as they have been identified in benign and borderline mucinous precursor neoplasms ,16. The Molecular testing is additionally performed in the clinical setting on an as-required basis in cases that pose diagnostic challenges, most notably in the sex cord stromal tumours. Almost all adult granulosa cell tumours harbour a recurrent somatic FOXL2 point mutation , whilst A further subset of ovarian tumours has molecular alterations described in the literature; however, these are not required for diagnosis and have not as of yet been proven to be efficacious in the clinical setting in terms of directing targeted therapies. For example, low-grade serous carcinomas with KRAS or BRAF mutations are associated with a favourable prognosis; however, these are not routinely tested . SimilarIt is, therefore, evident that the landscape for molecular testing in ovarian tumours is likely to change dramatically in the coming years as targeted therapies become available for as-of-yet unactioned tumour mutations. In an attempt to better organise molecular testing within the United Kingdom and to ensure equitable access to all patients, genomic testing is now delivered via seven Genomic Laboratory Hubs (GLHs), which are distributed throughout the country, with each hub responsible for coordinating the molecular diagnostic services for its own region. Requests for molecular testing are submitted via online forms to increase efficiency; however, not all tests are currently performed within the NHS. At present, HRD testing is outsourced to Myriad Laboratory in the United States. This has an impact on turnaround times (TATs), as diagnostic material must be sent abroad for testing. The intention is to ultimately consolidate all molecular testing services to within the NHS, which will have a significant impact on reducing TATs and, therefore, enhance patient outcomes through the rapid generation of molecular reports.In order to deliver a timely, streamlined, cost-effective, and accurate molecular diagnostic service, the pathologist plays a key role as an intermediary between the clinicians and those involved in molecular testing. The challenges posed by this process are plentiful and shall be discussed in detail below.There are multiple steps involved in the generation of a timely and accurate molecular pathology report; therefore, it is imperative that clear pathways for the communication of requests for tumour testing are jointly established between clinical teams and pathology services at both local and national levels. The processes involved in molecular testing of any tumour type can be broken down into pre-analytical, analytical, and post-analytical phases. The pre-analytical phase starts with tissue sampling and extends up to the point of molecular testing, which is the analytical phase. The post-analytical phase refers to the interpretation and communication of test results. The pathologist is involved in both the pre- and post-analytical phases, whilst the molecular testing is performed by biomedical scientists and biostatisticians. There are a number of variables present within the pre- and post-analytical phases that can be modified and optimised in order to ensure the greatest chance of generating accurate test results, as discussed below.In order to generate a robust and accurate molecular pathology report, a sufficient volume of viable tumour cells is required for testing. When a tissue sample is processed in the lab, sections are first cut to assess tumour morphology on a routine H&E slide. In many instances, further sections are required for ancillary testing with immunohistochemistry (IHC) and/or special stains, to confirm or refine the diagnosis. It is often not until this point that the remaining tissue can be sent for molecular testing, if required. It is, therefore, apparent that scanty biopsy samples are not likely to contain sufficient tumour cells to both accurately identify the tumour histotype and perform appropriate molecular testing. For this reason, it is strongly recommended that more than one tissue core is taken when performing diagnostic biopsy sampling. In order to preserve tissue for molecular testing, individual cores can be embedded into separate blocks, and a single H&E will be processed for each in order to assess morphology in the first instance . Based oIntratumoural heterogeneity plays an important role in influencing the results of molecular tests. Ovarian cancers are known to have high rates of heterogeneity, which refers to the fact that there is a phenotypically diverse population of cancer cells present both within a single tumour and across multiple different tumour subtypes . This diEach molecular test has a pre-defined limit of detection, which refers to the lowest concentration of analyte that can be detected and consistently generate an accurate test result. The analyte in question in the setting of molecular testing in ovarian cancer is the tumour cell, and the concentration is the number of tumour cell nuclei divided by the total number of nuclei present in a tissue sample. A high-quality sample should contain a percentage of tumour cell nuclei that measures at least double the stated limit of detection for the required test. In general, this corresponds to ≥30% tumour cell nuclei; however, >20% will often suffice. It is clear that tissue samples with large numbers of background benign cells, including inflammatory cells and stromal cells, will result in a reduction in the percentage of tumour cell nuclei present for assessment; however, steps can be taken by the pathologist to avoid this becoming an issue. By examining the morphology of each tumour sample on H&E prior to requesting molecular tests, it is possible to choose the sample with the greatest volume of tumour cell nuclei upfront to avoid inconclusive or failed test results. Similarly, one can macro-dissect an area of tissue with high tumour cell content away from the surrounding tissue and send this alone for molecular testing in order to reduce the impact of non-tumoural tissue on the molecular report generated.It is very difficult to macroscopically predict the tumour cell content of any tissue at the time of sampling; however, there are a number of factors to consider that may indicate a lower quality sample site. Many tumours will contain areas of necrosis, which can often be identified macroscopically. These areas are to be avoided when taking biopsy samples as they do not contain viable tumour cells. In the setting of percutaneous biopsy sampling, it may be impossible to avoid these areas, thus multiple biopsies should be taken to avoid the need for a repeat procedure . SimilarThere is a clear hierarchy in terms of the best type of material for molecular testing. Resection specimens that have not received NACT are theoretically ideal as they classically provide large volumes of viable tumour for testing; however, this is not often the tissue type chosen for analysis in light of both clinical need for a pre-operative tissue diagnosis and the frequency of standard neoadjuvant treatment regimens currently administered. A good-quality biopsy sample is often the preferred choice, as it may be used to direct delivery of neoadjuvant therapies. As previously discussed, it is imperative to take more than one biopsy at the time of sampling. The use of post-NACT tissue for analysis is often suboptimal, depending on the degree of response to treatment seen, as defined by the chemotherapy response score (CRS). A CRS score of 1 refers to no or minimal response to therapy; CRS 2 describes an appreciable response to therapy with readily identifiable residual tumour; CRS 3 is a complete or near complete response to therapy, with scattered residual tumour cells remaining at most . The greOnce a tissue sample has been taken from a patient, it is routine practice for it to be placed immediately into 10% neutral buffered formalin, following which it is processed and embedded in paraffin wax to create a formalin-fixed paraffin-embedded (FFPE) block . The optThe benefit of adequate tissue fixation prior to processing is that the tissues are preserved in as life-like a state as possible through inhibition of autolysis. This is of particular importance in the setting of surgeries or biopsy clinics held on a Friday afternoon: because most pathology laboratories do not operate a full service over the weekend, if a sample is not placed in formalin at the time of removal by the clinical team, it is possible that this will not be recognised over the weekend. In this case, the degree of natural autolysis that the tissue will undergo may lead to inadequate viable tumour cells available for both histological and molecular diagnostics. Similarly, as is often seen in institutions with pathology departments located remote from surgical theatres, delays in the transportation of specimens to the lab following retrieval may result in tissue over-fixation, with inherent potential for an impact on DNA quality.Not all material that has been formalin-fixed will reliably provide high-quality DNA for molecular analysis. In many instances, as briefly mentioned above, the DNA extracted from FFPE tissue is often fragmented and of poor quality as a result of the process of fixation and on a molecular level may contain artefactual sequence alterations resulting from crosslinking of protein molecules . NeverthWhen a tissue diagnosis that prompts further molecular testing has been made by a pathologist based on morphological appearances, the process of requesting molecular testing depends on the test type and indication. In the setting of HRD testing for HGSC, patients must first be consented by the clinical team, and this consent (or lack thereof) should clearly be stated on the histology request form or in another predetermined location agreed by those involved in this care pathway. Tests that are performed as part of routine diagnostics do not require such explicit consent. Following the identification of the most-appropriate tumour sample, tissue curls of a 6 μm thickness should be cut by the medical scientists in the pathology laboratory and sent along with the completed molecular request form to the appropriate GLH to which the referring hospital is connected. Providing tissue curls, as opposed to sections mounted onto glass slides, is preferrable as they result in less damage to tissues during DNA extraction because the tissues do not need to be scraped off of slides. It is worth noting that the preferences of different molecular laboratories may vary in terms of how they wish to receive and process the material; thus, conferring with the GLH prior to sending specimens is advised. The pathologist can help to expedite molecular testing, for example in the case of HRD, by requesting tissue sections or curls reflexively upon making a diagnosis of HGSC of stage 3 and above. Once patient consent has been confirmed, the lab can release this material to the GLH for testing. This process can be made yet more efficient by communicating patient consent via electronic patient records, rather than through emails between clinicians and pathologists.From this point on, the process of molecular analysis enters into the analytical phase, which is regularly performed by biomedical scientists and biostatisticians rather than histopathologists.Following molecular testing, the pathologist is furnished with a molecular report from the GLH, which needs to be interpreted in the clinical context of the individual patient and communicated to the clinical team in an efficient, clear, and concise manner. This is often a non-onerous task in the case of straightforward reports; however, it becomes more difficult when the results generated are more complex and require a greater level of understanding of molecular biology to interpret. It is important remember that most histopathologists working in the clinical setting do not have specific training in high-level molecular pathology. There is now a clear career path for those who wish to pursue this route, with FRCPath fellow status by examination; however, this route is normally pursued by medical scientists and biostatisticians, rather than clinical histopathologists. It is, therefore, imperative that a clear report is provided to the pathologist in the first instance so that there are no interpretive errors or delays in generating a report to send to the clinical teams.The time commitments involved in issuing molecular reports has also increased significantly in recent years, as both the availability of and indications for molecular testing have expanded rapidly. Previously, molecular testing in the case of HGSC was limited primarily to patients with a known significant family history, whilst at present, HRD testing is performed in every patient who provides consent as a positive result can guide the delivery of targeted therapies. This results in a far greater number of tests that must be requested, interpreted, and reported by the pathologist, all with their own inherent time commitments. Similarly, this increased testing imparts an economic impact on the pathology services. On a local level, many of these costs have been addressed by the establishment of national GLHs, which perform the tests; however, increased expenses relating to tissue processing, transport, and storage are generally absorbed by the local department.The use of molecular pathology in the clinical setting has changed dramatically in recent years and is set to transform yet again in the near future as our understanding of the molecular biology of ovarian tumours increases and as new targeted medical therapies are developed. Due to the significant potential impact on the treatment and broader management of individual patients and their families posed by the results of these tests, it is imperative that all steps are taken in order to ensure that the testing has the greatest chance of success. The pathologist is heavily involved in the process of molecular testing; however, as discussed above, their ability to provide high-quality tissue samples for testing is heavily influenced by various preanalytical factors. It is advised that clear pathways for communication of molecular test requests be established on a local and national level, with regular feedback between the teams regarding current issues with the process. The establishment of small changes to practice, as required, can have an enormous impact on the quality of test results provided and, therefore, greatly optimise the clinical outcomes for patients with ovarian tumours under our care.


Kaposi sarcoma (KS) is a low-grade mesenchymal angioproliferative disorder that requires infection with human herpes virus 8 (HHV-8) for it to develop. It is commonly seen in HIV-positive patients and rarely in immunosuppressed HIV-negative patients. Rituximab is a monoclonal anti-CD20 chimeric murine/human immunoglobulin G antibody used to treat B cell lymphoproliferative diseases as well as a variety of autoimmune disorders. Several cases of iatrogenic Kaposi sarcoma (iKS) have been described after rituximab treatment.The purpose of this narrative review is to identify the presence of common clinical characteristics among rituximab-induced KS patients that could facilitate better management of this rare condition. Kaposi sarcoma (KS) was described for the first time in 1872 by Moritz Kaposi. He observed several cases of cutaneous multifocal pigmented sarcoma , HIV-assCurrently, there are four clinical variants of KS, each with distinct natural history, prognosis, and site propensity: (i) classic KS mainly affects the extremities of the lower limbs of elderly men in Mediterranean countries and the Middle East; (ii) the epidemic subtype is associated with acquired immunodeficiency syndrome (AIDS-associated KS), which mainly affects men who have sex with men infected with the human immunodeficiency virus (HIV); (iii) iatrogenic subtype occurs in patients treated with immunosuppressive therapy, especially organ transplant recipients; (iv) endemic subtype is observed primarily in sub-Saharan Africans . IatrogeRituximab is a monoclonal anti-CD20 chimeric murine/human immunoglobulin G antibody used to treat B cell lymphoproliferative disease . Its mecRituximab therapy has been reported to exacerbate HIV-associated KS, most likely by immune system suppression in already immunocompromised HIV patients. To date, rituximab-related KS in HIV-negative patients has been reported by only a few articles in the literature. Rituximab may thus have a role in HHV-8 lytic replication and facilitate the development of KS in the absence of HIV infection. The goal of this narrative review is to provide an overview of the role of rituximab in the development of KS in HIV-negative patients (KS-nHIV).MethodsA literature search evaluating the role of rituximab in the development of KS was conducted using the National Library of Medicine (MEDLINE), the Excerpta Medica database (EMBASE), and the Cochrane Database of Systematic Reviews (CDSR). The literature search was conducted between May 2023 and June 2023. The inclusion criteria consisted of English-language articles, case series, and case studies that reported on the role of rituximab in the development of KS. We examined the title and abstract of each identified study, and the full text of the study was consulted for relevant studies. The following keywords were used in the search: Kaposi sarcoma, iatrogenic Kaposi sarcoma, and rituximab-induced Kaposi sarcoma. A flowchart describing article selection is included in Figure Pathogenesis of KSHHV-8, commonly known as Kaposi sarcoma-associated herpes virus (KSHV), is a major contributor to the pathogenesis of KS ,16. ChanThe role of HIV and KSHV coinfection in the development of KS cannot be understated . For exaRole of rituximab in KS developmentRituximab is the first monoclonal antibody drug used in oncology. It was approved by the Food and Drug Administration in 1997. The first licensed indication was for the treatment of relapsed/refractory indolent non-Hodgkin lymphoma. Since its approval, rituximab’s therapeutic indications were extended and presently include numerous other B-cell malignancies, autoimmune diseases, inflammatory disease, paraneoplastic pemphigus, pemphigus vulgaris, hypocomplementemic urticarial vasculitis, autoimmune cytopenias, and humoral organ transplant rejection. Rituximab is also prescribed for multicentric Castleman’s disease and primary effusion lymphoma ,25.Rituximab is a human/mouse chimeric monoclonal antibody (IgG1) that targets the CD20 antigen expressed in more than 95% of normal and malignant B cells and induces complement-mediated and antibody-dependent cellular cytotoxicity. It has been reported as a triggering factor for KS . RituximThe main targets of HHV-8 are B cells and endothelial cells, thus B cells, due to their long life, can be major latent reservoirs of HHV-8. Activated CD4+ T cells are capable of suppressing virus production by these infected B cells. In contrast, a major decrease in the number of CD4+ T cells is considered a significant risk factor associated with KS and multicentric Castleman’s disease, whatever the cause of the T cell-dependent immunosuppression .Lower B cell counts were also associated with a statistically significant increased risk of KS development, while higher B cell counts were protective against KS development. These data highlight a potential role for B cells and the humoral immune system in KS pathogenesis Some authors believe that a rituximab-related decrease in T-cell activation plays a key role in HHV8 reactivation. Although rituximab was a groundbreaking drug in the treatment of B-cell lymphomas, rituximab-associated activation of hepatitis B and HHV-8 are adverse effects that delay chemotherapy. The risk-benefit ratio of rituximab, especially for non-lymphomatous disorders, such as autoimmune and systemic inflammatory disorders, needs to be carefully assessed for patients at risk of developing HHV-8 tumors . The rolRituximab-induced KS in patients treated for non-Hodgkin lymphomaThe number of cases of patients developing KS after treatment with rituximab for hematologic malignancies is limited in number, consisting mainly of case reports.A case study by Billon et al. reports on a KS case that developed under rituximab treatment for follicular lymphoma in an HIV-negative, 55-year-old patient . The les2, vincristine 2 mg, cyclophosphamide 750 mg/m2 on day 1, doxorubicin 50 mg/m2 on day 1 and methylprednisolone 80 mg for five days, every three weeks) [Another case report by Alkan et al. describes the case of a 65-year-old treated for stage IIA B cell lymphoma with an R-CHOP regimen (rituximab 375 mg/me weeks) . After 1Ureshino et al. described a similar case in an older HIV-negative patient with numerous comorbidities who was treated with an R-CHOP regimen for diffuse large B cell lymphoma (DLBCL) . After cAnother case is that of a 52-year-old male patient with a history of mantle cell lymphoma treated with rituximab, dexamethasone, cytarabine, cisplatin/oxaliplatin , and autologous stem-cell transplantation. Five months after treatment initiation the patient noticed several cutaneous lesions on the nasal alae and ear lobes. Skin biopsy confirmed the KS diagnosis and tumor cell nuclei stained positive for HHV-8. Rituximab treatment was stopped and the patient remained in follow-up. After one year, the KS lesions have remained stable .2 was initiated for KS. The KS lesions exhibited partial response after two months of vinblastine therapy; however, the patient later died due to complications unrelated to KS or lymphoma.A case report by Uyanik et al. describes the case of a 69-year-old male patient with splenic marginal zone lymphoma treated with R-CHOP regimen with partial response . The patRituximab-induced KS in patients treated for chronic lymphocytic leukemia2 and day 1 and day 2 bendamustine 60mg/m2. After 18 weeks of rituximab treatment, the patient developed a series of papular lesions on both lower extremities. After the biopsy, the diagnosis was established as KS with positive HHV-8 staining. Doxorubicin 20mg/m2 was initiated and within three cycles there was a significant lesion response.A case report by Belur et al. showcases the case of a 76-year-old HIV-negative female patient diagnosed with chronic lymphocytic leukemia (CLL) . The patA retrospective study by Brambilla et al. included 1389 KS patients of which 143 were suffering from iKS . PatientSimilarly, another case report describes the case of a 70-year-old male patient diagnosed with CLL and treated with six cycles of rituximab + fludarabine + cyclophosphamide. After a short interval, a purple lesion appeared on the left foot of the patient. The diagnosis of HHV-8-positive KS was established following the biopsy. The patient underwent radiation at a total dose of 30 Gy with the disappearance of the skin lesion after three months .Rituximab-induced KS in organ transplant recipientsAs seen in the previous section, organ transplant recipients (OTR) had a higher chance of developing KS compared to patients who were immunosuppressed for other reasons.A retrospective study by Périer et al. included 988 patients that received rituximab and only four male HIV-negative, HHV-8 tumor-positive patients developed KS, at a median of four months after the first RTX infusion . One 56- A 2010 retrospective study by Scemla et al. reported on the complications associated with the use of rituximab in renal transplant patients . Out of Rituximab-induced KS in patients with autoimmune diseaseKS was described in several cases of autoimmune disease treated with rituximab. A case report describes a 52-year-old male HIV-negative patient diagnosed with thrombotic thrombocytopenic purpura (TTP) that was initially treated with corticosteroids. After initial improvement in platelet count, he exhibited severe thrombocytopenia that was treated with four weekly doses of rituximab at 800 mg per dose. After therapy completion, he developed several purple lesions on his upper and lower extremities. These lesions were diagnosed as KS and treated conservatively with liquid nitrogen cryo-ablation. At follow-up, the patient had no recurrence of TTP or KS .Another report by Daflaoui et al. describes the case of an 82-year-old male with pemphigus vegetans that was treated with a total of two doses of rituximab of 1000mg every two weeks . Three mSimilarly, a case report by Torretta et al. describes the case of a 68-year-old male patient with pemphigus vulgaris who was treated with rituximab as well as corticosteroids and immunoglobulins . After cAnother case of rituximab-induced KS was reported in a 76-year-old female patient with stage IVb (according to the Myasthenia Gravis Foundation of America staging) myasthenia gravis that associated bulbar and respiratory manifestations. After first-line therapy with pyridostigmine and glucocorticoids which proved ineffective, rituximab treatment was proposed. After serology testing, the possibility of acute or chronic HBV, HCV, and HIV infection was excluded. Six months after rituximab induction, several purple/blue skin nodules appeared on the trunk, arms, and legs and were later confirmed as KS. Rituximab was stopped. After staging the work-up using thoracic-abdominal-pelvic computed tomography, several suspect lesions were identified in both lungs. The patient was diagnosed with stage IV KS and started on pegylated liposomal doxorubicin (PLD). Complete resolution of the nodular skin lesions was observed after the first two PLD administrations. Following six cycles of PLD, disease progression was noted and, due to poor performance status, chemotherapy was stopped and the best supportive care was initiated. The patient died 14 months after rituximab initiation .2 was proposed and after the second infusion, a complete response was observed. Before the last dose of rituximab, the patient developed nodular lesions on both legs. After a skin biopsy, the diagnosis of KS was established followed by three courses of oral etoposide 60mg/m2. After completion of etoposide treatment, the lesions disappeared and no recurrence was noted at follow-up [ In the final case described in the literature, a 79-year-old man diagnosed with autoimmune hemolytic anemia secondary to chronic lymphocytic leukemia received treatment with methylprednisolone, which proved ineffective. Treatment with rituximab 375mg/mollow-up .Management of rituximab-induced KSEven though no widely accepted guidelines on the management of iKS exist to date, reducing or discontinuing immunosuppressants represents the most accredited therapeutic option in iKS . This thLocalized treatment may consist of surgical excision, radiotherapy, intralesional chemotherapy, silver nitrate cauterization, and compressive therapy with elastic stockings. Systemic treatment can consist of various drugs: liposomal doxorubicin, sirolimus (for transplant KS), paclitaxel, pomalidomide, bortezomib, gemcitabine, lenalidomide, vinorelbine, etoposide, imatinib, ipilimumab + nivolumab, pembrolizumab.The mechanisms of rituximab-induced KS are largely unknown. Rituximab-induced B-lymphocyte depletion determines latent HHV-8 reactivation and implicitly favors the development and progression of KS . SecondaAlthough lytic viral replication may play a role in some aspects of KS, antiviral therapy is not likely to be of benefit when used as monotherapy for KS, as the role of persistent viral replication after malignant transformation is unclear . We specDiscussionThis narrative review aims to establish if rituximab can be responsible for KS development. While data in the literature are scarce and mostly consist of case reports several pertinent conclusions can still be made. The series of case reports included in this review describe KS occurring in individuals who have been treated with rituximab, particularly when rituximab is used in combination with other immunosuppressive therapies. This fact raises the question of whether rituximab alone can be considered a risk factor for KS development. To answer this, we must look back at the mechanism of rituximab-induced KS.The depletion of B-cells by rituximab can weaken the immune system's ability to control viral infections, including HHV-8, which is a key factor in the development of KS . AdditioIt is essential to note that rituximab itself does not directly cause KS. Instead, it may increase the risk of developing KS in individuals who are already infected with HHV-8 or who have other risk factors, such as a compromised immune system from either a different cancer or in HIV-positive patients from acquired immunodeficiency syndrome. Therefore, patients receiving rituximab and other immunosuppressive therapies should be carefully monitored for signs of KS and other opportunistic infections, especially if they have underlying conditions that make them more susceptible to these complications.Despite the extensive use of rituximab in the treatment of a variety of ailments, the reported case number of secondary KS in HIV-negative patients is extremely low. To our knowledge, approximately 30 cases have been described so far in the literature  Table 11. While The average time from rituximab initiation until KS development was 6.8 months (1.5-30 months). All patients benefited from complete immunosuppressive therapy or just rituximab therapy cessation. With no clear guidelines for iKS management, the subsequent KS treatment varied widely, from complete resection of the lesions to systemic therapy and radiotherapy. That being said prognosis is quite favorable irrespective of treatment with most patients exhibiting complete or partial response during follow-up. Only one patient which was included in this review died during follow-up with an overall survival of 11 months.Even though iKS is a rare occurrence, caution is required, especially for cancer patients who are frequently immunocompromised by both their condition and their treatment. That being said some key aspects of managing rituximab-induced KS in HIV-negative patients are: 1) discontinuation of rituximab, 2) KS-specific treatment (the choice of treatment depends on the extent and severity of the KS lesions), 3) management of immunosuppression , 4) regular monitoring  and HBV (hepatitis B) infection during rituximab treatment as well as other potential opportunistic infections, and 5) preventive measures .Several limitations of this review include the lack of data available in the literature, and most of the articles described being case reports. We believe that high-quality studies performed on larger patient populations should be conducted so that clear guidelines can be developed. That being said it is important to emphasize that the management of rituximab-induced KS is complex and should be tailored to the individual patient's specific medical history and condition. Our review highlights a noteworthy association between rituximab therapy and the development of KS in HIV-negative patients. While rituximab is an effective treatment for various medical conditions, including autoimmune diseases and certain malignancies, it appears to be a potential risk factor for KS. The role of rituximab-induced immunosuppression in the pathogenesis of KS cannot be overlooked, although data is limited and only several studies report on this association. This review also suggests that not all patients receiving rituximab are at equal risk for KS. Certain patient-specific factors, such as underlying medical conditions  and previous immunosuppressive therapies, may contribute to varying susceptibility. Timely diagnosis and monitoring of KS are paramount, especially in patients undergoing rituximab treatment. Close clinical surveillance and vigilant evaluation of skin lesions, oral mucosa, and other potential sites of KS lesions are crucial for early intervention. We have also shown that the management of rituximab-induced KS involves a multifaceted approach, including discontinuation of rituximab, KS-specific treatment modalities, and tailored adjustments to immunosuppressive regimens for autoimmune conditions.


Mpox and MARV viruses.The increasing incidence of Monkeypox virus (Mpox) and Marburg virus (MARV) infections worldwide presents a significant challenge to global health, as limited treatment options are currently available. This study investigates the potential of several O-rhamnosides and Kaempferol-O-rhamnosides as Mpox and MARV inhibitors using molecular modeling methods, including ADMET, molecular docking, and molecular dynamics/MD simulation. The effectiveness of these compounds against the viruses was assessed using the Prediction of Activity Spectra for Substances (PASS) prediction. The study’s primary focus is molecular docking prediction, which demonstrated that ligands  bind to Mpox (PDB ID: 4QWO) and MARV (PDB ID: 4OR8) with binding affinities ranging from -8.00 kcal/mol to -9.5 kcal/mol. HOMO-LUMO based quantum calculations were employed to determine the HOMO-LUMO gap of frontier molecular orbitals (FMOs) and to estimate chemical potential, electronegativity, hardness, and softness. Drug similarity and ADMET prediction assessments of pharmacokinetic properties revealed that the compounds were likely non-carcinogenic, non-hepatotoxic, and rapidly soluble. Molecular dynamic (MD) modeling was used to identify the most favorable docked complexes involving bioactive chemicals. MD simulations indicate that varying types of kaempferol-O-rhamnoside are necessary for successful docking validation and maintaining the stability of the docked complex. These findings could facilitate the discovery of novel therapeutic agents for treating illnesses caused by the  In the middle of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection that emerged as a novel virus in 2019, and has since caused the ongoing coronavirus disease 2019 (COVID-19). Two additional zoonotic viruses, Monkeypox virus (Mpox) virus and Marburg virus (MARV), have recently re-emerged in the current years . With ovThe viruses are obligate intracellular parasites that can act as living, non-loving objects . They arAccording to the recent outbreaks and findings, viruses are still the cause of a vast variety of infectious illnesses in humans and animals, as well as a substantial proportion of newly developing and reemerging infections that are considered to be of significant health concern . Some emin vitro, such as Adamantane derivatives, Resveratol, Tecovirimat, Mitoxantrone, and Ribavirin. Nevertheless, their usefulness and safety in the context of Mpox have not been shown, and further basic and clinical research is needed  was used to predict the anti-bacterial, anti-fungal, and anti-parasitic spectrum (http://www.swissadme.ch)  . These padme.ch) .https://www.rcsb.org/) which were isolated by X-ray diffraction method with high stable configuration. After that, PyMol version 2021 was applied for removing all the excess heteroatom and minimized the energy by Swiss PDB viewer , and MARV (PDB ID: 4OR8) were download from Protein Data Bank (PDB) , it is revealed that the L02, L03 and L04 are formed by attaching one, two and three O-rhamnoside rings with the original compound of O-rhamnoside, as shown in In computational chemistry, the use of quantum mechanical methods for the calculation of thermodynamic, molecular orbital, and molecule electrostatic characteristics is prevalent. The program known as Gaussian 09 has been run in order to improve the geometry and further enhance each generated derivative. The density functional theory (DFT), functional of B3LYP and Gaussian double zeta plus polarization function basis set (DNP), was utilized in order to improve and predict the molecular orbital and thermal properties of the molecules. All compounds had their dipole moment, enthalpy, free energy, and electronic energy determined. The optimized molecular structures are shown in The probable biological spectrum for Kaempferol-o-rhamnoside derivatives has been predicted by applying the web server PASS. The PASS data is summarized as Pa and Pi, which are shown in Due to a variety of factors, the most medication candidates never become commercially available or cannot pass the clinical or preclinical stages. It is the utmost significance to create trustworthy computational approaches for the estimation of the drug-likeness of new drug candidates that increase the percentage of successful drug discovery and development attempts during the trial phase. The drug-likeness prediction model was developed by Christopher A. Lipinski in 1997, which included molecular descriptors connected to the numbers of factors that can differentiate between possible drugs and non-drugs. These factors include topological polar surface area, number of rotatable bonds, hydrogen bond acceptor, hydrogen bond donor, and molecular weight. The molecular weight of the reported Kaempferol-o-rhamnoside derivatives (L01-L09) was at 164.16-754.69, the number of rotatable bonds was 0.0-10, the hydrogen bond acceptor was 05-19, the hydrogen bond donor was 04-13, and the topological polar surface area was 90.15Å² -318.37Å². So, only Ligand of L01, L02, L06, and L07 satisfied the overall Lipinski rule  are the most important orbitals in a molecule. They are used by scientists to study chemical reactivity and kinetic stability. People often talk about the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) when talking about these molecular orbitals on the edge of the atom (LUMO). When you say “electronic absorption,” you mean a change from the ground state to the first excited state. This is usually thought of as one electron moving from the HOMO state to the LUMO state . A molecAs indicated in From the The Molecular Electrostatic Potential, also known as MEP, is utilized as a global reactivity map. This map illustrates the amount of an organic molecule that is most equipped for the electrophilic and nucleophilic onslaught of charged point-like reagents . It is hThe forces between molecules that occur between the ligand and the receptor during the formation of a protein-ligand complex are called binding affinity. This method has grown in popularity as a result of the research and investigation of innovative drug design . A bindiThe Kaempferol-o-rhamnoside derivatives generated a wide variety of different types of non-covalent interactions with the active sites of the microbial proteins. These observed bonds included pi-alkyl, pi-anion, pi-sigma, amide pi-stacked, and pi-donor hydrogen bonds. Based on these results, it is abundantly obvious that aromatic substituent may quickly enhance the binding ability of uridine esters as well as their potential to inhibit the growth of reported viral pathogens due to the high electron density of aromatic substituent. The influence of H-bonds had a significant impact on the selectivity of ligand binding to the receptor, drug design in chemical and biological processes, and molecular recognition and biological activity. Therefore, the H-bond surface of all derivative ligands that interact with both proteins is presented in Molecular dynamics (MD) simulations are effective tools for comprehending the physiological underpinnings that encompass the structure and function of bioactive molecules . The iniSecondly, the MD simulation has been completed for ligands 08 and 09 towards the MARV (PDB 4OR8). In this case, the RMSD and RMSF have been obtained with equivalent outcomes, and the RMSD is about 0.9 Å – 1.2 Å both time vs. backbone and backbone with amino acid residues, while the RMSF score at 0.7 Å – 0.8 Å  of protein. From the The present system of advanced manufacturing drug development includes the use of in silico techniques as an essential element. These tools are used to explore the absorption, distribution, metabolism, and excretion (ADME-PK) features of novel chemical substances. So, this ADMET analysis was performed to examine both the pharmacokinetic (PK) features and the toxicity of the compound. Kaempferol-O-rhamnoside derivatives (L01-L09) have been studied for their potential as agonists to suppress Mpox and MARV infections; these compounds have been modeled using computational tools using DFT method, as well as molecular docking, molecular dynamics simulation, and in silico study.st four ligands (L01-L04) which is more reactive, and it is slightly lower for next class that is about 7.0 or below. However, the softness of L01 –L04 is about 0.21 where the softness for other is about 0.25 to 0.27. However, the 2nd series of ligand (L05-L09) after joining is more active in case of docking but chemical more stable where L07, L08 and L09 show higher binding score. So, it can be said that with increasing the chain length, the binding capacity increases.In addition, investigations are conducted on pharmacokinetics, ADMET, drug-likeness, HOMO/LUMO gap, molecular electrostatic potential, and PASS prediction in an effort to offer them with an effective antiviral drug against Mpox and MARV infections. With beginning, the PASS prediction investigation outcome obtained at 0.379< Pa<0.712 for anti-viral (Influenza), 0.423< Pa<0.655 for anti-bacterial, 0.632< Pa<0.755 for antifungal, and 0.214< Pa<0.391 for anti-parasitic, so it can be said that they have a high possibility against viral pathogens. Next, molecular docking has been described as the most important part of this investigation, and it has achieved an acceptable docking score, which was even higher than the FDA-approved anti-viral Cidofovir drug. In accumulation, there are formed the hydrogen bonds after docking that conveys the more stability of protein as inhibitor. All of designed drugs showed the negative value of BBB permeability, CYP4501A2 inhibitor, CYP4502C9 substrate and Renal OCT2 substrate even they have no AMES toxicity. In case of quantum calculation, the energy gap is about 9.00 for 1The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding authors.AK, NM- Conceptualization, study design, writing, data acquisition. MA- Doing the simulation, DFT calculation and data analysis. AC, SM, AG, UC, SA, AK, AAla- Writing and data acquisition. AD, RS, AG, AAle and K-TC – Review, editing, and supervision. All authors contributed to the article and approved the submitted version.


In the striatum,presynaptic α6-containig nicotinic receptorsare crucially involved in the modulation of dopamine release. CVN417,a novel selective antagonist at this receptor subtype, attenuatesmotor dysfunction in a Parkinson’s disease-relevant animalmodel, suggesting, for this pathology, a therapeutic strategy thatcould greatly profit from the restricted localization of α6*nicotinic receptors in the brain. Within BG circuitry, the striatumis the major processing unit. About 95% of striatal neurons are GABAergicprojections neurons, named medium spiny neurons (MSNs). The remaining∼5% of striatal neurons are about half-and-half GABAergic andcholinergic interneurons (ChIs) (2Parkinson’sdisease (PD)is classified among the 18 major neurological disorders globally,with its rank of disease burden moved up from 14th in 1990 to 10thin 2019.s (ChIs) 1.22 In a recent issue ofthe Journal of Medicinal Chemistry, the Bürligroup at the Cambridge drug discovery company Cerevance reported anovel brain-penetrant α6-containing nicotinic receptor antagonist,CVN417 are selectively localized 1.2 In a ,CVN417 2,that m1 receptors (D1Rs) and promoting movement, and the indirectpathway, terminating ongoing movement and predominantly influencedby inhibitory D2 receptors (D2Rs).2 A striatal dopamine (DA) deficit is classicallythought to underlie many of the movements disorders in PD becauseit results in reduced activation of the direct pathway and reducedinactivation of the indirect pathway.2l-DOPA, the first drug of choice when treating PD, is used tocounteract dopamine deficiency inasmuch as it is the dopamine precursorcapable of crossing the blood–brain barrier.Modulatory dopaminergic afferents act through twomain pathways,referred to as the direct pathway, mediated by excitatory D2Rs on striatal ChIs, which activates the indirect pathwaysvia M1Rs on MSNs. ChIs are a small fraction of all striatalneurons, but they have a great influence over striatal motor outputs,and they express a multitude of receptor subtypes, which could allowACh transmission to be modulated, though not without unwanted sideeffects. Therefore, normalizing ChI overactivity remains, to date,a pursuable goal for the treatment of PD at disease stages when ChIsare still intact.2Imbalanceddirect and indirect pathway signaling is accentuatedby elevated striatal ACh, resulting from reduced activation of D2Rs on ChIs lowers ACh levels, while blockingM1Rs on MSNs attenuates the indirect pathway. However,even these approaches are not without drawbacks and adverse effects.4Targeting muscarinicreceptors is an alternative therapeutic option.Stimulating M4 Their prevalent localizationis on DAergic terminals, where α4β2*- and α6β2*-nAChRsare expressed and seem to act by depolarizing the terminal boutonand producing dopamine release.5 In contrast to the α4β2*subtype, which is widespread throughout the brain, the α6β2*subtype has a relatively selective localization to the nigrostriatalpathway, restricted, moreover, in the striatum to DAergic terminals.Such a unique distribution makes the α6β2*-nAChR, morethan the α4β2* subtype, an outstanding target to treatdisorders linked to the nigrostriatal system.4Within this context, striatal nicotinicACh receptors (nAChRs)can play an untapped distinctive role as viable targets to improveparkinsonian state. Notably absent, unlike M and D receptors, fromMSNs, striatal nAChRs result from different combinations of α4,α6, α7, β2, and β3 subunits, with a predominanceof α6β2- and α4β2-containing receptors (α4β2*-and α6β2*-nAChRs).6 However, althougha number of successive studies highlighted the importance of the striatallocalization of α6β2*-nAChRs and their peculiar involvementin motor behaviors, the results have remained elusive. First of all,non-decisive or conflicting evidence about reduction of PD incidenceand alleviation of PD motor symptoms arises from the experiments withboth chronic and acute dosing of nicotine, a non-selective nAChR agonistdirectly effecting on the midbrain dopamine.2 Second, beneficial effects of nicotinic agonists targeting bothα4β2*- and α6β2*-nAChRs are documented in L-DOPA-induced dyskinesia but not in PD models.7 Third, administration of a moderately α6β2*-selectiveagonist, such as TC-2429 , andantagonism (mecamylamine and dihydro-β-erythroidine) serveto discriminate tonic and phasic patterns of stimulation enhancingthe sensitivity of dopamine release to burst versus non-burst stimuliand that nicotinic agonists achieve the same effect as antagonists.These processes would be mainly mediated by presynaptic α6β2*-nAChRs.This notwithstanding, in the past two decades,accumulating evidenceon the importance of presynaptic location of α4β2*- andα6β2*-nAChRs has prompted a series of studies highlightingthe nature and the impact of the modulation of dopamine release byacetylcholine, nicotine, and nicotinic drugs acting at these nAChRssubtypes. In a low-frequency tonic mode, dopamine neuron activitymaintains low nanomolar concentrations of dopamine, while bursts ofhigh-frequency phasic activity produce much greater release of dopamine.When dopamine neurons are quiescent (tonic firing), nAChRs are tonicallyactive, responding to a physiologically pulsatile delivery of ACh,and their desensitization is avoided. Instead, experimentally inducedhigh concentrations of ACh produce desensitization and decrease dopaminerelease at low-frequency stimulation.3 is the actualization of such a potential strategyfor the treatment of PD symptoms by engaging in the development ofan α6*-nAChR-selective antagonist, which should be a more accessibleachievement than development of a selective agonist. The researcherssucceeded by screening a library of ∼650k compounds for inhibitoryactivity at the α6-containing nAChR. Selection of the most promisinghit was followed by a relatively streamlined hit optimization, resultingin the development of CVN417 as a lead molecule. Indeed, CVN417 combineshighly potent and selective α6-containing nAChR antagonism,determined in vitro and ex vivo, with high efficacy in attenuatingresting tremors in an animal PD model and, on the basis of in-depthprofiling by ADME and DMPK assays, good pharmacokinetic properties.Furthermore, evidence is provided that CVN417 would exert its effectsin vivo, presumably by a neurotransmission regulation involving modulationof phasic DAergic neurotransmission in an impulse-dependent manner.The major highlight of the study by Bürli’s groupThis research provides a novel tactic that could be applied inPD therapy. Further studies will prove whether this is, indeed, apracticable option. Additionally, it will be interesting to studythe impact of selective α6-containing nAChR antagonism alsoon the significantly problematic non-motor symptoms associated withPD.


Streptococcus pneumoniae (SP) antimicrobial resistance (AMR) and the CDC/ATSDR Social Vulnerability Index (SVI) in the US.Antibiotic resistant infections affect more than 2.8 million people in the US each year and disproportionately impact those who are socially vulnerable. The objective of this study was to evaluate the association between Adult patients (≥ 18 years) with 30-day nonduplicate SP isolates from ambulatory/hospital settings from January 2011–December 2022 who had zip codes of residence reported were evaluated across 177 facilities in the BD Insights Research Database . Isolates were identified as SP AMR if they were intermediate or resistant to ≥ 1 macrolide, tetracycline, extended-spectrum cephalosporin, or penicillin. Isolates from adult patients with zip code of residence reported were matched with county-level CDC SVI data, and associations between SVI score  and SP AMR were evaluated using generalized estimating equations with repeated measurements within county to account for within-cluster correlations .P=0.0002; Table 1). Similarly, a single decile increase in household characteristic theme ranking score was observed to be associated with a 0.81% increased risk in SP AMR . However, no association between the racial and ethnic minority status theme, the housing type and transportation theme, and overall SVI score and SP AMR were observed.Of the 8,008 unique SP isolates linked to 574 US counties, the overall rate of SP AMR was 49.9%. A significant association between the socioeconomic status (SES) theme and SP AMR was detected with higher SVI scores  associated with greater risk of AMR. On average, a single decile increase in SES theme ranking score was observed to be associated with a 1.28% increased risk of AMR  were observed to be associated with higher risk of SP AMR. These results contribute to evidence linking AMR to health disparities and inequities and suggest that AMR is impacted by social determinants of health.Salini Mohanty, DrPH, MPH, Employee of Merck & Co., Inc.: Stocks/Bonds Gang Ye, PhD, Becton Dickinson and Company: employee Charles Sheets, MS, Becton Dickinson: Data Analysis Nicole Cossrow, PhD, Merck: Stocks/Bonds Kalvin Yu, MD, FIDSA, BD: Stocks/Bonds Meghan White, PharmD, Merck: Employee|Merck: Stocks/Bonds Kenneth P. Klinker, PharmD, Merck & Co., Inc: Employee|Merck & Co., Inc: Stocks/Bonds Vikas Gupta, PharmD, Becton, Dickinson and Company (BD): Employee of BD|Becton, Dickinson and Company (BD): Stocks/Bonds