Pancreatic ductal adenocarcinoma (PDAC) has still a dismal prognosis, mainly because only 15–20% of all patients present with resectable tumor stages at the time of diagnosis. Due to locally extended tumor growth or distant metastases upfront resection is not reasonable in the majority of patients. Considerably, PDAC will be the 2nd most frequent cause of cancer-related deaths within the next 10 years for both men and women. While there is currently no convincing evidence for the use of neoadjuvant therapy in resectable PDAC, there are controversial results from studies investigating neoadjuvant treatment concepts in borderline resectable PDAC (BR-PDAC). However, the definition of BR-PDAC is a topic of debate. While BR-PDAC has originally been defined on merely anatomical criteria, the International Association of Pancreatology (IAP) has recently suggested a broader definition based on a combination of anatomical (A) findings, biological (B) criteria (which reflect tumor aggressiveness), and conditional (C) aspects (which respect host-related condition). In case of BR-PDAC with venous invasion alone, upfront resection is generally recommended whenever technically possible in patients fit for surgery and without evidence for lymph node metastases. In contrast, in case of arterial invasion neoadjuvant therapy is regarded as the treatment of choice. The same accounts for high CA 19-9 levels, suspected or proven lymph node involvement and poor performance status. In locally advanced PDAC (LA-PDAC), neoadjuvant treatment represents the standard of care resulting in proportionally high rates of secondary resection. This “conversion” surgery offers the chance for improved survival times in an otherwise palliative situation. Herein, we summarize the current evidence of different treatment strategies for pancreatic cancer with a focus on conversion surgery and the impact of neoadjuvant treatment in this setting. Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal tumor entities and—although being currently the 3rd leading cause for cancer-associated mortality in the United States—will be the 2nd leading cause of cancer-related death within the next 10 years. This frustrating development can mainly be explained by an increasing incidence and still limited treatment success , and secFrom the anatomical point of view, resectable PDAC is defined as a locally limited tumor extension without any tumor contact with the superior mesenteric (SMV) or portal vein (PV), and also without involvement of the celiac axis (CA), its main branches and the superior mesenteric artery (SMA). In case of venous involvement, the criteria for BR-PDAC are fulfilled if venous resection followed by reconstruction is technically feasible. Cavernous collateralization of the portal venous axis toward the hepatic hilus or infiltration of the jejunal vein branches are findings of venous tumor involvement without the possibility of venous reconstruction and thereby fulfill the criteria of LA-PDAC, with the particularity that conversion surgery is rarely possible in these two scenarios despite neoadjuvant treatment. Arterial BR-PDAC is defined as an abutment below 180° of the SMA or isolated tumor contact to the hepatic artery. In addition to the situations described above, LA-PDAC comprises tumors with a more extended involvement of the SMA, CA, common hepatic artery, aorta, or inferior vena cava . Figure Besides the aspect of technical resectability, tumor biology must be considered independently from local tumor extension as there are patients who may have a localized and resectable, but biologically very aggressive tumor which implies that they have a high risk of local tumor recurrence or metastases very early after surgery. Unfortunately, today the diagnostic tools to preoperatively characterize the aggressiveness of the individual patient's tumor are highly limited as no precise markers have been available so far. Since years, the best clinically established marker for PDAC has been carbohydrate antigen (CA) 19-9 which gives a rough estimate on tumor biology although cut-off values to differentiate highly aggressive from moderate or low aggressive tumors are rather unreliable. In addition, ~15% of all patients do not express CA 19-9 at all, and in case of obstructive jaundice the interpretation of the CA 19-9 level is also very limited. The International Association of Pancreatology (IAP) was the first to consider tumor biology in their current definition of borderline resectability which was published 2 years ago . BesidesPatients fit for surgery with a resectable tumor should undergo surgical exploration and radical resection followed by adjuvant chemotherapy as long as convincing evidence for neoadjuvant treatment in this situation is lacking . ConsequNeoadjuvant treatment followed by conversion surgery represents a potentially beneficial treatment alternative for BR-PDAC and is currently evaluated in numerous studies including patients with either type of BR-PDAC  , 12. MosThese two examples of RCTs show two aspects that make a valid conclusion on the actual value of neoadjuvant therapy for BR-PDAC very difficult. First, most of the current studies have included very heterogeneous patient populations . Second, there is no international standard on neoadjuvant protocols at the moment so that institutions around the world chose their preferred treatment including chemotherapy, chemoradiation, or a sequential treatment pre- and postoperatively with both modalities. In contrast to this, the standards of adjuvant therapy after upfront resection are clearly defined on the basis of large multicenter studies making this concept an evidence-based approach.Regarding LA-PDAC, neoadjuvant therapy, and conversion surgery have undergone a considerable development in recent years. While venous LA-PDAC is characterized by a technically unreconstructable venous tract, arterial LA-PDAC is basically technically resectable but in general considered as unfavorable from an oncological point of view as it may reflect an extremely aggressive tumor biology. Consequently, venous LA-PDAC is unlikely to become technically resectable after neoadjuvant treatment, whereas arterial LA-PDAC has to be regarded differently.In this context, tumors of the pancreatic body which involve the CA represent a special situation in which a distal pancreatectomy with transection of the CA and the hepatic artery under preservation of the gastroduodenal artery can be performed if the arterial perfusion of the liver is ensured by retrograde blood from the SMA . Preferably, DP-CAR is embedded in a neoadjuvant setting as shown in most recent studies. A cohort study from Johns Hopkins included 17 patients with PDAC of the pancreatic body undergoing DP-CAR following neoadjuvant treatment and matched them with 51 patients undergoing DP only . In the Besides the above mentioned scenario allowing DP-CAR, arterial LA-PDAC has historically been regarded as a palliative situation and arterial resections in PDAC surgery have only been performed in few patients in the past , 24. ThiModern neoadjuvant therapy has changed the picture of arterial resection being an only highly-individual approach and it has been shown that effective neoadjuvant protocols can considerably increase secondary resection rates despite an unresectable tumor stage at the time of diagnosis. This development can be referred to two main potential effects of neoadjuvant therapy: (1) down-sizing and maybe down-staging of the tumor, and (2) devitalizing the tumor without an obvious imaging-based response which allows to spare arterial resections in many patients during conversion surgery. The Japanese Society of Hepato-Biliary-Pancreatic Surgery published a series including 58 patients with initially unresectable PDAC who underwent 8 months of neoadjuvant treatment followed by “adjuvant resection” which resulted in clearly improved survival times compared to a palliative approach, illustrating the important role of neoadjuvant treatment in conversion surgery .P = 0.026). The median overall survival after resection was 15.3 months compared with 8.5 months after exploration alone (P < 0.0001). When considering the neoadjuvant treatment period, further 5–6 months of survival are to be added (The evaluation of tumor response to neoadjuvant therapy remains an unsolved problem in the treatment of LA-PDAC because a clearly radiographic response can be seen in only approximate one third of the patients . There ibe added . The Heibe added . In patibe added  can be pbe added . In manybe added . Comparebe added .Recent meta-analyses on the efficacy of neoadjuvant treatment in patients with LA-PDAC confirmed the benefit of pretreatment, primarily with FOLFIRINOX, but also revealed low evidence for the use of neoadjuvant chemotherapy and chemoradiotherapy owing to the limited methodological quality of the original publications available , 40. OneConsidering the limited suitability of conventional imaging, diagnostic tools to better evaluate response to neoadjuvant treatment would be desirable as well as biomarkers to better select patients who will benefit from a multimodal treatment approach. Although no clear cut-off values for CA 19-9 are defined, however, the relative drop of CA 19-9 after neoadjuvant therapy can give additional information on potential resectability and oncological prognosis , 42. In In summary, the paradigm of regarding LA-PDAC as a non-surgical and palliative disease is undergoing substantial changes today. Different approaches of neoadjuvant therapy show encouraging results underlining the importance of a multimodal strategy to improve these patients' prognosis. Standardization of neoadjuvant approaches, however, remains poor to date. Still the aim of conversion surgery in LA-PDAC can be achieved in a considerable number of patients and effective chemotherapy protocols with or without radiation are the key to further enhance these results and should be investigated in prospective clinical trials to allow evidence-based recommendations in the future.TH and UK conceived and wrote this review article.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Scanning electron microscopy has been developed for topographic analysis at the nanometer scale. Herein, we present a silicon p-n diode with multi-annular configuration to detect backscattering electrons (BSE) in a homemade desktop scanning electron microscope (SEM). The multi-annular configuration enables the enhancement of the topography contrast of 82.11 nA/μm as compared with the commercial multi-fan-shaped BSE detector of 40.08 nA/μm. Additionally, we integrated it with lateral p-n junction processing and aluminum grid structure to increase the sensitivity and efficiency of the multi-annular BSE detector that gives higher sensitivity of atomic number contrast and better surface topography contrast of BSE images for low-energy detection. The responsivity data also shows that MA-AL and MA p-n detectors have higher gain value than the MA detector does. The standard deviation of measurements is no higher than 1%. These results verify that MA p-n and MA-AL detectors are stable and can function well in SEM for low-energy applications. It is demonstrated that the multi-annular (MA) detectors are well suited for imaging in SEM systems. The scanning electron microscope (SEM) has been widely applied to study surface topology and the shape of nano-objects in many research fields, ranging from semiconductors  to nano-x-axis) is preferable for topography reconstruction in SEM. However, the multiple detectors system requires a larger space and higher cost.As compared with the SE detector, the BSE detector has more diverse applications. It is well known that BSE signals can show not only COMPO detail contrast but also TOPO variation ,7. EarlyRegarding inspiration from the multiple detectors, in this paper, we modify the traditional commercial MF BSE detector to have multi-annular configuration of detection areas  detector) which is with the same size as MF BSE detector. The MA BSE detector has a four semi-rings configuration of detection area  and MF BSE detectors was calculated to be 0.598 and 0.576 sr, respectively [A BSE detector collects backscattered electrons from the area where the sample interacts with the incident electron beam passing through the central hole of the BSE detector. The traditional commercial BSE detector usually is operated with four fan-shaped (MF) detection areas, as shown in l MF BSE a detectoectively .As shown in 2 film on both sides of the silicon wafer  of detector is the working area is coated with Al grids. A schematic diagram for the fabrication processes is illustrated in As shown in To show the MA structure does give a better surface topography contrast as compared with a MF structure, the surface topography (TOPO) contrast was first tested with MF and MA detectors. Then the subtile improvement in electrical performance (I–V curve), effective gain of detectors, sensitivity of atomic number, as well as image contrast for backscattered mode (COMPO) of SEM were tested with three detectors MA, MA p-n and MA-Al BSE detectors. Besides, the stability of our annular detectors has been tested in a desktop SEM. A and IB are signal currents at the highest and lowest points of the sample, and HA and HB are heights at the corresponding highest and lowest points of the sample. The BSE images recorded using MF and MA are shown in In the test of surface topography contrast, two types of BSE detectors, MF and MA BSE detectors were mounted in a SEM . We used BSE images recorded from a commercial MF BSE detector as a bench mark to quantitatively compare with that recorded using our MA BSE detector. The standard sample used for quantification of the TOPO contrast was a silicon patterned microstructures etched by RIE with width of 1.2 μm and depth of 1.0 μm, as shown in The basic diode characteristics of electrical performance for the MF, MA, MA p-n and MA-Al detectors were tested by measuring the I-V characteristics. We obtained typical I-V curves of detectors by multi-meter  and put the detectors in SEM chamber to make sure that without illumination, as shown in The effective gain G is given by the ratio of detector output current to dose of electron beam . To determine the effective gain, we exposed the MA-, MA p-n and MA-Al detectors to direct electron irradiation also in a SEM  and the incident electron beam current was measured by using a Faraday cup.input is measured from the current of the incident electron beam with a Faraday cup and the Ioutput is measured from the current in our detectors. It shows that the MA-Al detector has better performance for low-energy electron detection. At the electron beam energy of 0.5 keV, the measured electron signal gain of the MA-Al detector has an approximate performance which as compared to the boron-layer silicon photo-diodes, which have been done by other teams [er teams .A and IB are signal currents for a sample of different atomic number; ZA and ZB are corresponding atomic number of different elements.To determine the sensitivity of atomic number, we prepared a standard specimen which inlaid five polished materials . Three different, BSE detectors  were mounted in a desktop SEM, EM-100. The sensitivity of atomic number contrast was defined as ,(3)SensSignal and IBackground are the averaged intensity from the circled gold particle and the silicon substrate, respectively, while the σSignal and the σBackground are the standard deviation of these two parts, respectively. As shown in This experiment was also performed in a desktop SEM (EM-100). The BSE image contrast for three different BSE detectors  were quantified using a sample of mixed gold and polystyrene particles. The image contrast C can be quantified as ,(4)C=|IIn addition, another sample of pure gold particles is used for the quantitative comparison of different BSE detectors, as shown in In this study, we designed and fabricated a simple processing and low-cost silicon detector with MA configuration of detection area to enhance topography contrast for the application of BSE detectors in desktop SEM. The MA BSE detector can provide better surface topography contrast of 82.11 nA/μm which is compared with the commercial MF BSE detector of 40.08 nA/μm. Moreover, we integrated it with several other special processes to increase the performance of the BSE detector. Without biasing, the detector presented here has a dark current in the order of 150 pA and a detector gain of 62.5 at electron beam energy is 0.5 keV. The modified BSE detector has better sensitivity of atomic number and BEI contrast than the MA BSE detector for low electron energy detection (about 16% enhanced). Further enhancement in detector performance is possible by decreasing the width and pitch of the aluminum grids. Higher capability can be expected and it will become a powerful device in SEM systems.


Mycobacterium tuberculosis-host interactions. In this mSphere of Influence article, she reflects on how the paper “Single-cell phenotyping within transparent intact tissue through whole-body clearing” by B. Yang et al.  impacted her ideas on approaches to visualize and understand heterogeneous host-pathogen interactions in vivo in 3-dimensional space at the single-cell level, through the tractable and broadly compatible tissue optical clearing methods developed.Shumin Tan works in the field of  Mycobacterium tuberculosis-host interactions. In this mSphere of Influence article, she reflects on how the paper “Single-cell phenotyping within transparent intact tissue through whole-body clearing” by B. Yang et al.  impacted her ideas on approaches to visualize and understand heterogeneous host-pathogen interactions in vivo in 3-dimensional space at the single-cell level, through the tractable and broadly compatible tissue optical clearing methods developed.Shumin Tan works in the field of  In partin situ) procedure, with both antibodies and small-molecule dyes delivered via the vasculature and with the washes required between the optical clearance and antibody administrations also exploiting the circulatory system (A main principle behind tissue optical clearing lies in the removal of packed lipid bilayers that impede both light and antibody penetration –4. There–3–y system .Mycobacterium tuberculosis-host interaction studies that my group is focused on, heterogeneity across multiple facets of the infection has been of burgeoning interest due to the consequences of this nonuniformity for infection outcome and treatment success. This heterogeneity ranges from lesion-level progression and outcome to variation in the local environment experienced by single bacilli even within a given lesion or host cell (–Vibrio cholerae and Salmonella infection of the intestine to Yersinia pseudotuberculosis colonization of the spleen (–The technological advances developed by Yang et al., particularly the passive optical clearing method of PACT and the use of RIMS, excitingly represent a method that is straightforward and easily applied to any organ and that, importantly, is compatible with fixed tissue and downstream immunofluorescence analysis without the need for highly specialized equipment. Particularly in the context of ost cell –14. Indee spleen –17. Howe+ T cell visualization, in lymphoid tissues in humanized mice (Mycobacterium marinum-zebrafish model (M. tuberculosis responses in intact lung tissue, we have started to combine PACT with our reporter M. tuberculosis strains that inform on the local environment experienced by the bacteria, or on bacterial replication status, in a C3HeB/FeJ murine model that exhibits a range of lesion types, including classic caseating granulomas (Helicobacter pylori colonization establishment and persistence in the murine stomach (The paper by Yang et al. has been extensively cited since its publication in 2014. There has been additional work on methods for optical clearing of tissue, as well as application of the clearing methodology to various fields, spanning visualization of blood brain barrier permeability in studies of prion disease  to the ezed mice . Opticalsh model , as wellsh model . To examanulomas . Finally stomach . These s


Computational inference of affected functions, pathways and diseases for chemicals could largely accelerate the evaluation of potential effects of chemical exposure on human beings. Previously, we have developed a ChemDIS system utilizing information of interacting targets for chemical-disease inference. With the target information, testable hypotheses can be generated for experimental validation. In this work, we present an update of ChemDIS 2 system featured with more updated datasets and several new functions, including (i) custom enrichment analysis function for single omics data; (ii) multi-omics analysis function for joint analysis of multi-omics data; (iii) mixture analysis function for the identification of interaction and overall effects; (iv) web application programming interface (API) for programmed access to ChemDIS 2. The updated ChemDIS 2 system capable of analyzing more than 430 000 chemicals is expected to be useful for both drug development and risk assessment of environmental chemicals.https://cwtung.kmu.edu.tw/chemdisDatabase URL: ChemDIS 2 is freely accessible via  The identification of chemical-induced effects on human beings is vital for both the drug development and chemical risk assessment. The application of traditional experimental methods to fully characterization of chemical-induced health effects could take several years and require enormous resources for a single chemical. Due to the huge number of chemicals that may cause diseases, the use of high-throughput methods for chemical prioritization can greatly save resources and time. Various modern experimental approaches have been developed for this purpose. For example, large-scale omics projects such as DrugMatrix , CMAP 22, 3 and To address the need for fast analysis of chemical-induced effects, a computational system named ChemDIS for chemical-disease inference has been developed for analyzing >90 000 chemicals including many poorly characterized chemicals such as maleic acid and sibutramine , 7. The In this report, we present a new ChemDIS 2 system with four major improvements. First, the applicability has been extended to include >430 000 chemicals and >15 million chemical–protein interactions based on STITCH 5 . Second,The previous version of ChemDIS was based on the integration of R packages and related SQLite databases. While a Rserv server was implemented in ChemDIS to accelerate computational performance, it is no longer suitable for dealing with the largely increased dataset in ChemDIS 2. In this work, we have implemented all programs using GO language and a modern NoSQL database of MongoDB supporting the fast calculation, high-performance access to data and web APIs. Database and programs were deployed in an Ubuntu server.https://chun-weitung.gitbooks.io/chemdis/content/data-sources.html). Currently, >430 000 chemicals with >15 million chemical–protein interactions can be analyzed using ChemDIS 2 that is >3 times larger than the original ChemDIS system.To enable the inference of chemical-induced effects, several databases were downloaded and integrated into a MongoDB database including STITCH 5 , ReactomThe flowchart of ChemDIS 2 for single chemical analysis is shown in The user interface has been redesigned based on AngularJS providing better user experiences in ChemDIS 2 as shown in In addition to the major update to the search function for chemical-protein-disease association inference, ChemDIS 2 offers three useful tools to help the analysis of chemical-induced effects including functions for custom, multi-omics and mixture analyses. Instead of relying on chemical–protein interaction data from STITCH database, the custom analysis function accepts user inputs of a custom set of differentially expressed genes, proteins, miRNAs or metabolites that could be derived from various omics experiments, and enrichment analysis will be conducted. Significantly affected functions, pathways and diseases will be identified for the user-supplied set of gene, miRNA or protein identifiers. For example, our previous work conducted gene expression profiling of maleic acid-treated neuronal cells and identified differentially expressed genes whose associated functions were analyzed based on the custom analysis function . The serp-value ip represents the adjusted p-value for dataset i. The joint p-value has been shown to be effective for the identification of enriched terms supported by multiple datasets  for detailed information and examples of currently available web API. An example of web API is shown in While ChemDIS 2 is equipped with an easy user interface, there is an increasing need for programmatic access to core functions of ChemDIS for streamlining analysis workflow and implementation of new servers incorporating the analysis functions. To improve the interoperability of ChemDIS 2, a new web-based application programming interface (API) was implemented using Go language for accessing core functions such as the identification of interacting proteins and chemical-diseases associations. The web API is freely accessible via simple HTTPS calls. For example, given the PubChem CID of interest along with selected database version and a confident threshold for retrieving interacting proteins, the server will return a JSON (JavaScript Object Notation) object consisting of all analysis results. As an increasing number of functions are being developed, please refer to the online manual ; National Health Research Institutes (NHRI-107A1-EMCO-0318184); and Research Center for Environmental Medicine in Kaohsiung Medical University from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan. Open access publication was supported by Ministry of Science and Technology of Taiwan (MOST104-2221-E-037-001-MY3). The funding agencies play no role in the study design, data analysis and manuscript preparation.Conflict of interest. None declared.


Yet, such virtually barrier-less binding is only observed within a limited contact zone. Agglomeration hence leads to the formation of oligomers of nanoparticles, whilst full merging into a compact precipitate is hampered by the need for extended structural reorganisation. Implementing sufficiently fast supply from colloidal solution, our simulations show the development of silica networks comprised of covalently bound, yet not fully merged nanoparticles. Within the oligomerized nanoparticle network, coordination numbers range from 2 to 5 –which is far below closest packing. Our simulations hence rationalize the formation of covalently bound network structures hosting extended pores. The resulting interfaces to the solvent show water immobilization only for the immediate contact layers, whilst the inner pores exhibit solvent mobility akin to bulk water.The agglomeration of silica nanoparticles in aqueous solution is investigated from molecular simulations. Mimicking destabilization of colloidal solutions by full removal of protective moieties or surface charge, association of SiO In the past few decades, silica particles and nanocomposites thereof were used in many applications ranging from catalysts to coatings and reinforced plastics.–4 In addDespite such broad interest, our in-depth  understanding is rather limited. Size-controlled synthesis and functionalizing of silica nanoparticles for customizing properties calls for profound knowledge of the behavior in solution. Understanding and controlling interfacial interactions, association and aggregation is undoubtedly vital to the goal-oriented use of colloidal silica particles. This motThe aim of the present work is to outline a molecular simulation approach to colloidal silica association, ripening reactions and the formation of mesoporous precipitates. This calls for profound characterization of molecular interactions and the assessment of extended relaxation processes at the same time. To cope with the inherent time-length scale challenge to molecular simulation, we employ the Kawska-Zahn approach originally developed for studying crystal nucleation from solution.,20 While2/Si(OH)4 core/shell particles were prepared from a nanoblock (~cube of 38.5 Å dimensions) of alpha-quartz comprising 8 ×8×7 unit cells. Therein, the surface oxygen atoms were alternatively depleted / protonated. Using molecular dynamics simulations in vacuum, amorphous colloids were obtained from melting (imposing 2300 K) and rapid cooling to 300 K within 0.5 ns. The stability of the resulting nanosphere of ~ 5 nm diameter . For each possible ripening reaction, relaxed structures and energy levels of the reactant and product states are sampled from molecular dynamics runs using force-fields2+ → [SiO]2+ + H2O in the gas phase. The error margins of this approach may be estimated by shifting further neighbors from the MM to the QM treatment. This was found to change ΔEQM/MM by several tens of kJ/mol . Our QM/MM calculations hence aim at fast discrimination of exothermic/endothermic proton transfer attempts based on rough estimates with up to 20% error margins.The underlying subsystem should be chosen as small as possible to provide the computational efficiency required to scan hundreds to thousands of possible ripening reaction steps. In an earlier study of ZnO formation from zinc and hydroxy ions, proton transfer could be described by considering only the two reacting OHanically. For the The approaching of two silica colloids in water was first investigated from potential of mean force calculations using restraints on the colloid center-of-mass distances  to characterize the permeation of solvent shells and particle contact, respectively. For the All MD simulations in this study were performed with LAMMPS using a time step of 1 fs. Aqueous 2 core, whilst the surface comprises of silanol groups. The distribution of Si4+, O2- and OH- ions as functions of the distance from the center-of-mass is shown in - and O2-) hydrogen bonds with the silica particle. While it is intuitive to expect such strong electrostatic interactions to provoke well-defined solvation shells, the local roughness of the particle  prevents the formation of spherical solvent shells  in water, we find barrier-less binding of the two colloids. -∙∙OH- hydrogen bonds that may be subject to proton transfer reactions. Indeed, a total of 13 ripening reactions by exothermic proton transfer and structural relaxation could be observed  surfactants or surface charge to avoid agglomeration.,2 In turobserved , right. observed . Convergobserved . In lineobserved . This caDepending on colloid concentration in the solution, the association of further colloids will hence outperform the kinetics of ripening/merging beyond the colloid-colloid bonds described above. Experimentally, the underlying kinetics may be balanced by the choice of solvent, colloid concentration and by surfactant molecules to temporarily hinder the ripening of silanol groups.,34,35 Wi-∙∙OH- hydrogen bonds. Therein the overall number of reactive events is interpolated according to:In order to achieve a large number of such aggregation processes at reasonable computational costs, in what follows we mimic the (elsewise rather extreme number of) ripening reactions in an approximate manner. For this, the observed number of 13 ripening reaction steps is related to the contact area of the ripened dimer :To quantify the porosity of the resulting structures, we calculated the molecular surface of the silica agglomerate (i.e. the surface area accessible to solvent molecules) as a function of aggregate size N–with N being the number of colloids grown together , left. IOn this scale, the degree of porosity can be ‘quantified’ in terms of mixing the limiting functions to match the observed profile of A(N) see also . For the2/3 and N, respectively.[-1/3). While a proper quantitative assessment would call for larger aggregates, our simulation runs at least allow a qualitative identification of motif occurrence in the bulk as predominantly Nc = 2,3 and 4 –whilst a minor fraction of Nc = 5 is observed. To this end, we can rationalize the way that pores result from loops of connected colloids. We find 2, 3 and 4-fold coordination at roughly equal occurrence. While the former coordination numbers  would call for 2-dimensional aggregates, 3-D structures stem from Nc ≥ 4. We hence find a (roughly) 2:1 blend of 2D and 3D type connectivity for the colloidal building blocks that explains the formation of loops from filling 3-dimensional space by a large fraction of 2-dimensional motifs.The origin to this high level of porosity is the connectivity of the colloids forming its building blocks. We therefore performed a coordination number analysis of colloid-colloid contacts as a function of aggregate size . While iectively. Because + and Cl- we find poor association to the silica surface. The interface to silica is hence almost exclusively formed by water molecules which in turn experience significant reduction in mobility. This applies to two layers of solvation which in total extend over approximately 0.8 nm from the surface of particle. For quantification of the latter issue, we computed a 3-dimensional profile of ‘local’ water diffusion constants, by dividing space into a 0.2 nm sized grid and sampling the mean-squared displacement of water molecules for each of the grid points.[+/Cl- ions between the meso-porous interior and the embedding solution, respectively. It is intuitive to expect similar permeation also for small solute molecules–as suggested for pharmaceutics release from meso-porous silica vehicles.[To characterize the interplay of the meso-porous silica precipitate with penetrating solvent, we performed a large scale (6 ns) molecular dynamics run of the aggregate comprising 35 colloids see also . The sold points. The resuvehicles.Molecular simulations may bridge the scales from reacting ions to the agglomeration of nanoparticles leading to (largely) unprejudiced models of complex nanomaterials. Starting from individual association events and ripening reactions our simulation protocols unravel the nature of cavities and pores formation and provide atomic-scale details of surface/interface properties. Silica surely reflects a particularly prominent case study, however the modelling techniques are widely transferrable to characterizing meso-porous materials based on building-block association.While the present work reveals the foundations of porosity and solvent permeation, the resulting models paved the way to a large variety of future studies. These may involve the role of surfactants to control silica association and the use of colloids with disperse size distribution. Apart frThe latter perspective calls for much larger aggregates, and even 3D-periodic models of mesoporous silica. While impractical to provide from nano-particle by nano-particle association and ripening, we can still employ the beforehand discussed findings to create coarse-grained models that bridge the required time- and length scales. In ongoing efforts, we also introduce an account of additive molecules and their ability to form micelles that stabilize hollow structures as widely used in real syntheses of meso-porous silica. ,8,10,17S1 File(Table A) Van-der-Waals Interaction parameters. (Table B) Partial charges and Lennard-Jones parameters ion-water and water-water interactions.(DOCX)Click here for additional data file.S2 FileObtained Models of solvated single particle, solvated dimer particle and mesoporous particle.(ZIP)Click here for additional data file.


Crohn’s disease (CD) is associated with a variety of extra-intestinal manifestations. Most commonly these involve the eye, skin, joints, coagulation system and liver. Cerebral manifestations of CD have been reported to a far lesser extent. The extensive detrimental impact of neurological symptoms on a patient’s quality of life makes an early diagnosis and treatment particularly important. In previous case-reports, diagnosis of cerebral manifestations in CD often relied upon magnetic resonance imaging (MRI) and computed tomography (CT) alone. To our knowledge, only one case-report has documented a histologically confirmed case of cerebral lesions associated with CD so far.A 39-year-old right-handed woman with a history of CD was referred to our hospital with etiologically unexplained Gadolinium (Gd)-enhancing cortical lesions, triggering epileptic seizures. A CT-scan of the thorax and bronchoalveolar lavage found no signs of sarcoidosis. Lumbar punctures and laboratory testing found no underlying infection or coincidental autoimmune disorders and MRI-scans showed progression of lesion load. Consequently, the patient underwent stereotactic biopsy of a cortical lesion. Histological examination revealed a mixed lympho-histiocytic and tuberculoid granulomatous inflammation surrounding small vessels and no signs for infection. After exclusion of other granulomatous diseases and the typical histological findings we diagnosed a cerebral granulomatosis as a manifestation of CD. The patient was initially started on azathioprine, which had to be switched to corticosteroids and methotrexate because of an azathioprine related pancreatitis. The patient has not suffered any further epileptic seizures to date.Cerebral manifestation of CD is a possibly underreported entity that may respond well to immunosuppressive treatment. In contrast to earlier reports of cerebral manifestations in CD, our patient showed no coincident gastrointestinal symptoms indicating an activity of CD during the progression of cortical lesion load, suggesting that similar to other extra-intestinal manifestations in CD, the activity of gastrointestinal symptoms does not necessarily reflect the activity of CD associated cerebral vasculitis. Therefore, diagnosis and therapy of cerebral manifestation may be delayed when focusing on gastrointestinal symptoms alone. Crohn’s disease (CD) is an autoimmune disorder that primarily affects the gastrointestinal tract, but manifests itself in other organs as well . The mosThis 39-year-old right-handed female suffered from epileptic seizures that began to develop in early 2014. The epileptic seizures were initiated by dyscognitive seizures and secondarily evolved into tonic-clonic seizures. The patient was started on anticonvulsive medication in August 2014 (Lamotrigin and Levetiracetam). Under this regimen, which was adapted regularly, the frequency of seizures stabilized at about one seizure every six months.Some weeks prior to the first epileptic seizure, the patient had complained of novel subacute bifrontal headaches, which persisted intermittently over the course of the following months and were not directly correlated with the occurrence of epileptic seizures. Additionally, she reported a subjectively progressive impairment of her short-term memory. Her husband reported a temporary change in his wife’s personality during May 2015, which was accompanied by promiscuous behavior in social media and had not recurred since then.Gastrointestinal symptoms  had occurred for the first time in 2003. The diagnosis of CD was confirmed histologically via ileocolonoscopy in 2006. Symptoms were treated with a combination of steroids and mesalazine (5-ASA) from 2006 to 2008 and with methotrexate (15 mg once per week) from 2008 to 2016. The patient reported lack of gastrointestinal symptoms since 2013. Apart from nicotine abuse (25 pack-years) she reported no further comorbidities and did not suffer from any allergies. There was no family history of headaches, epilepsy, cancer and no exposure to toxic chemicals at home or at her workplace. There were no signs for lymphomatoid granulomatosis (no pulmonary and cutaneous lesions) or leukocyte oxidase deficiency (no recurrent infections). The patient did not travel outside of Europe in her entire life.The first isolated dyscognitive seizure was interpreted as a transient ischemic attack (aphasia) in early 2014 and the patient was started on aspirin 100 mg daily after a cerebral hemorrhage was ruled out via computed tomography. In August 2014 the first secondarily generalized seizure led to the initiation of a standard MRI of the head, showing supratentorial Gd-enhancing cortical lesions that were deemed ischemic or inflammatory in nature. A lumbar puncture revealed an inflammatory constellation . Bronchoalveolar lavage (including CD4/CD8-ratio) and a CT of the lungs were performed to test for (neuro-)sarcoidosis and provided normal results with no signs of perihilar lymphadenopathy. Furthermore, laboratory tests showed normal results for IL-2 receptor, ACE, neopterin, calcium, as well as for anti-neutrophil cytoplasmic antibodies (ANCA), anti-nuclear antibody (ANA) and anti-phospholipid-antibodies. An Interferon-gamma release assay test (QuantiFERON®) remained negative. A second MRI after two months (October 2014) showed further progression of the cortical lesions. Subsequently, the immunosuppressive therapy with methotrexate was complemented with steroids . A subsequent MRI in February 2015 showed a stable lesion load. Epileptic seizures in March, June and December 2015 as well as in March 2016 did not lead to adjustments of anticonvulsant or immunosuppressive therapy or to additional diagnostic measures.In April 2016 the patient was referred to our university center for epilepsy. Here, the dosage of the anticonvulsive medication was increased and another MRI was initiated (August 2016), which showed an increase of lesions with numerous bihemispheric, hemorrhagically imbibed lesions of the cortex, basal ganglia, midbrain and pons. Gadolinium-enhancing lesions were revealed in both corpora amygdaloidea and cortically in both inferior frontal gyri, the left frontal medial gyrus, the left postcentral gyrus as well as in temporo-occipital and occipital regions in both hemispheres. Additionally, the MRI showed bilateral hippocampal atrophy and hyperintensity  tests for Herpes simplex viruses 1 and 2, Varicella-zoster virus, tropheryma whipplei and a HIV-test, which remained negative. An ultrasonography of the extra- and intracranial arteries of the neck and brain showed no signs of a vasculitis of the large vessels. The biomarkers for sarcoidosis  remained negative, as did proteinase3 antineutrophil cytoplasmic antibodies (PR3-ANCA), perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) and blood cultures. There were no abnormalities in the differential blood count and in the immunoglobulins. A fluorescein angiography of the retina was performed in the department of Neuro-ophthalmology to test for a small-vessel vasculitis and Susac’s syndrome and revealed normal results.In September 2016 the patient was referred to our department of neurology via our colleagues at the university center for epilepsy in order to assess cerebral vasculitis. We performed another lumbar puncture  including antibody-tests for After completion of above-mentioned diagnostics, an interdisciplinary consent was reached to perform a stereotactic biopsy of one of the cortical lesions. Histology revealed leptomeningeal mixed mononuclear round cell infiltrates and multifocal mixed lympho-histiocytic and tuberculoid granulomatous lesions surrounding small vessels confirming the diagnosis of a cerebral granulomatosis Fig. -h. HistoOn the basis of the histological analysis and negative tests for infectious causes we diagnosed a cerebral granulomatosis as a manifestation of Crohn’s disease in the central nervous system and began immunosuppressive treatment with azathioprine, which had to be switched to methotrexate and corticosteroids because of an azathioprine related pancreatitis. To date, the patient has not suffered any further epileptic seizures and revealed no signs of cognitive deterioration in the outpatient follow-ups.Cerebral manifestation of CD is a possibly underreported entity , 12, 13,


Multiparametric Magnetic Resonance Imaging (mp-MRI) is the current standard of reference for the local staging of prostate cancer (PCa). On the other hand, despite the low sensitivity and specificity of Technetium Bone Scanning (BS) for the detection of bone metastases (BM) and of Body Computed Tomography CT for the detection of lymph node metastases (LNM), these techniques are routinely used, in the current clinical practice. Nevertheless, whole Body MRI (WB-MRI) and Positron Emission Tomography Computed Tomography (PET-CT) are emerging as robust tools for the staging of oncologic patients, including those with (PCa).The available techniques  for the detection of BM in oncologic patients were compared and showed striking center differences in terms of anatomic sequences and planes used. This heterogeneity and the long acquisition time of WB-MRI protocols – due to the addition of multiple anatomic sequences in different planes – questioned whether a single three dimensional (3D) sequence could replace the multiple anatomic sequences used for node and bone staging of PCa. We demonstrated that WB-MRI is a credible tool for the detection of bone and node metastasis.The second question addressed the possibility to obtain a complete TNM staging of PCa in a single MRI session. A WB-MRI protocol was developed to enable complete, T , N  and M (distant) staging of PCa in a single session, in less than an hour. This ‘all-in-one’ protocol proved to be as efficient as the sum of exams currently in use for the staging of PCa . Prostate cancer (PCa) is the most common non-skin cancer in America . AccordiMen with first-degree relatives with PCa have an increased risk for developing the disease . FurtherThe majority of men with PCa will remain asymptomatic. The symptoms – if they appear – are caused by the local extension of the disease and include dysuria, haematuria and erectile dysfunction. Sometimes the disease is detected only after a complication has occurred due to the metastatic spread, such as pathological fracture.Two tests are currently widely used for the screening of PCa: the digital rectal examination (DRE) and the blood Prostate Specific Antigen (PSA) levels testing. The increase of PSA levels can be associated with benign conditions, such as prostatitis and prostate hyperplasia. The risk of PCa for PSA values <0.5 ng/ml is 6.6% and it increases to 26.9% for PSA values between 3.1 and 4.0 ng/ml  whole-body MR images . Whole-bThe first studies utilizing WB-MRI were published between 1997 and 2001 2324. WB-232526The main disadvantages of WB-MRI are its relatively long scanning times, motion artefacts  and limited specificity. However, advances in hardware and imaging techniques, including additional sequences, are reducing the impact of some of these challenges. Most importantly, the introduction of DWI radically improves the diagnostic effectiveness of WB-MRI and promotes the technique as a tool of choice for PCa staging.Accurate distant and local staging of PCa patients is crucial in order to adapt treatment to the actual stage of the disease. Patients with bone metastasis cannot receive local treatment (surgery or radiation therapy), hence accurate detection of bone disease is essential. The development of clinical trials evaluating novel compounds and local treatment in the so-called oligo-metastatic patients has rendered the accurate evaluation of the metastatic burden indispensable 3031.30For N and M staging CT and/or abdominal MRI and Tc 99m BS with targeted X-rays (if necessary) are the most widely proposed staging methods 3334.33Additionally to its poor sensitivity and specificity, discussed above, the aforementioned multimodality algorithm necessitates multiple hospital visits and appointments in different departments (radiology and nuclear medicine), leading to additional times and discomfort for this group of patients (who are mostly elderly with multiple comorbidities). The delays for the interpretation of all these examinations also have to be considered.Multimodality algorithms would advantageously be replaced by one step modalities (WB-MRI and PET) protocols which have suggested their superiority and allow the rapid assessment of total tumor and all organ evaluation .18F-FDG that is a non-specific tumor-seeking radiopharmaceutical. The use of 18F-FDG is based on the heightened glycolysis in tumor cells, which provokes a high uptake of 18F-FDG by the tumor. Unfortunately PCa cells have low glycolysis levels and as a result they are not 18F-FDG-avid. This fact renders FDG PET modality poorly efficient for the detection of PCa metastasis . Our results demonstrated that TNM staging of PCa is feasible in less than one hour in an MRI session combining mp-MRI and WB-MRI, outperforming the routine screening for discriminating patients with or without metastasis.


Despite its high reactivity, the strong interaction of the radical’s SOMO with the LUMO of the cation frequently results in a high regioselectivity. Intra- and intermolecular processes such as the Minisci reaction, the Porta reaction, and the Knabe rearrangement will be discussed along with transition metal and photoredox catalysis or electrochemical methods to generate the odd-electron species.Carbon-centered radicals represent highly useful reactive intermediates in organic synthesis. Their nucleophilic character is reflected by fast additions to electron deficient C=X double bonds as present in iminium ions or cationic heterocycles. This review covers diverse reactions of preformed or  Radicals, representing odd-electron species, possess the peculiarity of combining nucleophilicity and electrophilicity at the same reaction center since their HOMO and LUMO  are located in one common molecular orbital, the SOMO . As a consequence, radical reactions are less influenced by Coulombic attraction than polar reactions but are rather governed by orbital interactions to a large extent. Thus, there is an analogy between radicals and “soft” electrophiles or nucleophiles for which orbital interactions are also primarily responsible for controlling their chemical reactivity .Being typically located in energy between the HOMO and the LUMO of the reaction partner, the overlap of the radical’s SOMO with both frontier orbitals will be energetically favorable. Depending on the SOMO energy, either the SOMO-HOMO or the SOMO-LUMO interaction may be dominant as the result of a smaller energy difference. In the former case, the radical can be classified as electrophilic while nucleophilic properties can be ascribed to it in the latter case  1]..1].vide infra). Usually, carbon-centered radicals are viewed as nucleophilic species with their nucleophilicity increasing with the degree of alkyl substitution or the presence of adjacent lone pairs as in the case of alkoxymethyl or hydroxymethyl but this of course strongly depends on the nature of the particular interaction partner. Thus, even radicals with an estimated electrophilic character are actually capable of reacting as nucleophiles if a reactant with a low lying LUMO is present. This behavior is an important aspect of the characteristic reactivity of radicals, making them a powerful, yet sometimes less predictable tool for organic synthesis.Carbenium ions and carbon-centered radicals share the same tendency of increased stability from primary to tertiary carbons which can be interpreted in terms of increasing hyperconjugative stabilization -orbitals, tertiary alkyl radicals have a higher SOMO energy than their secondary or primary counterparts and show an increased nucleophilic behavior  for the more electron-deficient 4-cyano derivative increases from 46 for the methyl radical over 1,300 for the sec-butyl radical to 350,000 for the tert-butyl radical  .,30.29,30et al. demonstrated the versatility of their reaction on various heteroaromatic scaffolds which further underlines the general applicability of Minisci-type reactions  and tert-butylhydroperoxide has been reported by Baran in 2011 . Th. Th37]. in situ generated benzylic radicals to iminium ions in 1984. The negative results of EPR and CIDNP were explained by efficient chain reactions. Proof for a free radical chain reaction was the formation of corresponding bibenzyls .,39.38,39ibenzyls  as side ibenzyls . The iniThe Knabe rearrangement is not limited to a benzyl substituent on C‑1 or on the isoquinoline moiety. It has also been observed with naphthalen-1-ylmethyl-, naphthalen-2-ylmethyl- or anthracen-9-ylmethyl substituents . Furtheret al.  from hexabutyldistannane to the ic cycle .8 s−1 . Op. Op99]. N-acylhydrazones were used by the Friestad group and the radical addition proceeded smoothly and with excellent selectivity  process. The generated odd electron species can react in different ways. Besides the unproductive backward reaction, a C–C bond fragmentation can occur to furnish an iminium ion and a neutral radical. Alternatively, deprotonation affords an α-amino radical, which can react with an olefin or can be converted to an iminium ion by another single electron oxidation. The same iminium ion can also be obtained by homolytic hydrogen abstraction from the initially generated amine radical cation. The resulting iminium ion reacts with nucleophiles to α-substituted amines and a radical addition is also possible  113]..113].et al. in 1992. 1-Ethylpyrrolidine and allyltrimethylsilane react with dicyanonaphthalene (DCN) and light irradiation to 2-allyl-1-ethylpyrrolidine (The radical addition to an iminium ion in a photoredox reaction had been discussed by Pandey rolidine . ThereinN-cyclopropylanilines with alkenes demonstrates the modes of reactivity outlined in The ruthenium-catalyzed photooxidation of mono- and bicyclic In summary, the strong interaction of the SOMO of carbon-centered radicals with the low-lying LUMO of a positively charged C=N double bond, as present in iminium ions as well as protonated or N-alkylated nitrogen heterocycles, results in fast and often highly selective addition reactions. This elementary process is the basis of a variety of several preparatively useful C–C bond forming reactions that generally do not require the use of protecting groups and can, for example, be used for the post-functionalization of complex substrates. Due to its efficiency, the radical addition to organic cations is likely to be part of further interesting developments in preparative free radical chemistry.


The EXP replaced their normal gymnastics physical preparation with a neuromuscular training program, while the CON continued with their habitual gymnastics program. Chi square analysis showed that the EXP resulted in significantly more positive responders compared to CON for measures of leg stiffness (41% versus 12% responded positively), extensor muscular endurance, (76% versus 29%), and competency in the deep overhead squat, (76% versus 29%) and in-line lunge (left lead leg) (65% versus 18%). Conversely, the number of positive responders for RSI (53% versus 61%), the flexor endurance test (88% versus 53%), and the right in-line lunge (47% versus 35%) were not significantly different between groups. These findings suggest that most young gymnasts responded positively to neuromuscular training from the perspective of improving movement proficiency and trunk endurance; however, changes in leg stiffness and RSI were more variable and may require higher intensities to realise further adaptations.This study examined individual responses in leg stiffness, reactive strength index (RSI), movement proficiency (deep overhead squat and in-line lunge), and trunk muscular endurance (flexor and extensor tests) in young female gymnasts following an 8-week neuromuscular training intervention. Thirty-four pre-peak height velocity (PHV) female gymnasts were divided into either an experimental group (EXP  Female artistic gymnastics is an early specialisation sport, typically involving high volumes and intensities of training during the pre-pubertal years aimed at mastering the performance of complex skills . Since rGymnastics involves a series of rebounding activities which utilise various expressions of stretch-shortening cycle (SSC) activity, ranging from slow-SSC  to fast-SSC activity  . The spoRecent research suggests that supplementary neuromuscular training  targeting the development of muscular strength could improve gymnasts’ ability to utilize their SSC , when peWhile research has demonstrated that youth can respond positively to forms of neuromuscular training ,11,20,26Unfortunately, very few studies in youth have examined the individual responses to neuromuscular-based training programs. One recent study found that in school-aged boys, individual responsiveness to training was influenced by both the mode of resistance training and maturity status . Howevern = 17 experimental group and n = 17 control group) volunteered to take part in this study. All gymnasts were from the same gymnastics club and were assigned to either the experimental (EXP) or control (CON) group based on the days they attended gymnastics training . Participant characteristics for both EXP and CON groups are presented in Thirty-four female artistic gymnasts aged between 6 and 12 years , and body mass using scales to the nearest 0.1 kg . Standing height, seated height, and body mass were entered into a validated sex-specific prediction equation  to deterThe EXP group followed an 8-week neuromuscular training program, consisting of two weekly sessions that lasted approximately 35 min each, which replaced their normal gymnastics conditioning. The CON group continued with their usual gymnastics program and did not receive any formalized strength and conditioning provision. The gymnastics programme for the CON group involved 35 min of gymnastics-specific conditioning , which was delivered by a club gymnastics coach and not the principle researcher. All testing sessions took place at the local gymnastics club where the participants trained. Both EXP and CON groups completed the same battery of tests at the beginning and end of the 8-week program (pre- and post-testing occurred 1 week either side of the program), and the same researcher administered each protocol. Gymnasts performed all tests barefoot wearing a gymnastics leotard. After a standardised 10-min dynamic warm up and practise trials, gymnasts performed the test battery in the following order: deep squat and in-line lunge (left and right lead leg), sub-maximal hopping, drop jump, and trunk endurance holds. All testing protocols have previously been shown to be reliable within the paedatric literature ,31,32. TParticipants were screened using a modified version of the FMS , whereby−1) was determined via a sub-maximal hopping test collected on a mobile contact mat . The Equation (A1) for leg stiffness can be found in Leg stiffness  from a box height of 20 centimetres . ParticiTo assess trunk muscular endurance, each gymnast performed the Biering-Sorenson test to assess trunk extensor and flexor endurance , isometrTo ensure technical execution of exercises was maintained throughout, the program was carefully supervised by the lead researcher who was a certified strength and conditioning specialist and qualified gymnastics coach. Program design adopted an integrative approach to neuromuscular training based on previous training interventions shown to be effective with inexperienced youth ,10,18,38t-test.Descriptive statistics (means ± standard deviations) were calculated for all pre- and post-data for each group. A multivariate analysis of covariance (MANCOVA) was used to control for baseline differences in group anthropometrics  for each performance variable. The assumption of normality was assessed via the Shapiro-Wilk test. A 2 × 2 repeated-measure analysis of variance (ANOVA) (group × time) was then used to examine the group changes in performance for each variable. Sphericity was assessed using Mauchly’s Test and where violated, a Greenhouse-Geisser adjustment was implemented. Percentage change from baseline testing was calculated for all individuals in each of the performance variables. The group percentage changes in performance were examined for each variable using an independent p < 0.05. Median values and percentages of individual scores for the deep overhead squat and in-line lunge data were calculated via Microsoft Excel for Mac version 15.35.In order to examine the individual responsiveness to the training intervention, the smallest worthwhile change (SWC) was calculated as 0.2 of the between subject SD for the total sample, using pre-intervention data. The SWC was expressed as a percentage of the group mean and a frequency count was then used to establish the number of individuals who made changes greater than the SWC, identifying those individuals who responded positively in performance. Chi-squared analysis was used to evaluate between-group differences for the number of positive responders for each performance variable. Analysis of the standardized residuals was used to identify frequencies that would be considered larger in magnitude than might be expected by chance , and wasp < 0.05). The between-group results from the repeated-measures ANOVA and percentage change are displayed in p < 0.05). Neither group showed significant improvements in RSI; however, the control group showed a significant reduction in leg stiffness.The results from the MANCOVA did not reveal any significant between-group differences for any performance variable at baseline testing when maturity was controlled for as a multivariate factor (n > SWC) for the flexor and extensor endurance test, leg stiffness, and RSI is shown in n > 1.96). Specifically, for leg stiffness, the number of positive responders and non-responders were greater in the EXP group (41% and 24%) versus the CON group (12% and 12%); while the EXP also showed a lower number of negative responders (35% versus 76%). A similar pattern of positive responders, non-responders, and negative responders emerged for extensor muscular endurance .The individual percentage change and the number of individuals that responded positively  compared to the CON . Conversely, the opposite pattern was evident for RSI .The median values and the distribution of test scores for the deep squat and in-line lunge are presented in The aim of this study was to examine individual responses in leg stiffness, reactive strength index (RSI), movement proficiency (deep overhead squat and in-line lunge), and trunk muscular endurance (flexor and extensor tests) in young female gymnasts following an 8-week neuromuscular training intervention. The main finding of this study was that an 8-week neuromuscular training program resulted in a higher percentage of responders in in pre-pubertal female gymnasts in most performance variables. The EXP group had a higher number of positive responders in both trunk endurance tests, the deep overhead squat and the in-line lunge tests, and had a significantly greater number of positive responders in leg stiffness compared with the CON. The number of positive responders for RSI were not significantly greater in either group. These findings suggest that supplementary neuromuscular training can provide additional benefits to pre-pubertal female gymnasts’ training programs. However, given the range of positive responders, non-responders and negative responders across all performance variables, alternative training prescription  may be required to promote more homogenous improvements in reactive strength and stiffness qualities.The group results from the deep squat and in-line lunge tests suggest that the training program was effective in eliciting improvements in the EXP group, with median values improving from a score of 1 to 2 in the deep squat and in-line lunge (consistent on both left and right limbs). While the distribution of the CON group’s individual scores showed improvement, the median values of the pooled data remained the same across all three tests. Participation in gymnastics can develop a variety of motor skills , but thiIn terms of the extensor and flexor muscular endurance tests, the EXP group made significant improvements in both tests, when compared with the CON. Previous research has shown participation in gymnastics develops trunk muscular endurance ; howeverExamination of the group mean data showed that the EXP group made non-significant improvements in leg stiffness, while the CON groups’ leg stiffness significantly decreased. Furthermore, neither group of gymnasts made significant improvements in RSI. These results indicate that on a group level, while the neuromuscular training failed to make significant improvements in performance, it was at least in part able to maintain leg stiffness and reactive strength qualities during the competitive period, where cumulative fatigue and detraining are more likely to occur. Previous research in elite pre-pubertal gymnasts showed gymnasts were unable to maintain gains in drop jump performance (from previous plyometrics and resistance training), confirming detraining following a competitive and transition period . TherefoWhile the group results provide some insight into the effectiveness of the neuromuscular training intervention, they fail to differentiate between gymnasts that were positive responders, non-responders, and negative responders. In terms of the individual response to training for movement proficiency, a greater number of individuals in the training group responded positively in all three tests compared with the CON group, and the total number of positive responders was significantly higher in two (deep overhead squat and left in-line lunge [left leg lead]) of the three tests. Interestingly, 14/17 of the individuals in the EXP group were right-leg dominant, which could indicate that the neuromuscular training was effective in starting to address lower-limb asymmetry that could have developed due their sports-specific training ,26. ThesExamination of the individual responsiveness for the extensor muscular endurance test determined that the neuromuscular training intervention resulted in significantly more positive responders (76%), when compared to the CON (29%). Interestingly, the distribution of positive responders, non-responders and negative responders for the flexor muscular endurance in the EXP group  suggests that the EXP group responded more favourably than the CON. However, the number of positive responders in the EXP group was not significantly higher than in the CON. Similar results have previously been noted in mature female gymnasts, which investigated the effects of a 10-week trunk muscular endurance training program (two 15-min sessions per week) on reducing incidences of low-back pain. The study showed that the training group significantly improved lateral flexor, extensor and flexor endurance, while the CON group only demonstrated improvements in flexor endurance . CumulatIndividual responsiveness data revealed that significantly more gymnasts in the EXP group elicited positive adaptations in leg stiffness (41%), when compared to controls (12%). However, when examining the number of positive responders versus the number of trivial and negative responders in the EXP group, the results indicate that the training stimulus only promoted improvements in leg stiffness for approximately 50% of gymnasts. Individuals who demonstrated improvements in RSI were not significantly greater than the expected count in either group. Furthermore, the distribution of positive responders, non-responders and negative responders for RSI  suggests that the addition of the current neuromuscular training program did not provide a superior stimulus to the gymnastics training alone. The differentiated individual responsiveness among both EXP and CON groups highlights the unique aspects of the sport of gymnastics, whereby the inherent physical demands of training and competing are likely to promote a degree of adaptation in various indices of SSC function .Previous research has shown that pre-pubertal gymnasts are superior in jumping tasks when compared to young athletes from other sports . A trainSome limitations should be noted in this study. Firstly, participant characteristics between the two groups were significantly different at baseline, with the CON group slightly more mature by approximately 1 year. However, a MANCOVA revealed no significant differences between the groups’ performance variables at baseline. Whether this difference in biological maturity would have influenced the post-testing findings despite both groups being pre-PHV is unclear and warrants further investigation. Secondly, as the current study took place over a competitive phase for the gymnasts, it is possible that accumulated fatigue influenced the results by blunting the training response for the gymnasts in the EXP group. Future research should aim to monitor workloads using rate of perceived exertion scales in both CON and EXP groups that are specific to youth . FurtherThe results from this study show that a neuromuscular training programme can provide additional benefits to some pre-PHV gymnasts, beyond gymnastics-training programs alone. With just 2 × 35-min sessions per week, strength and conditioning coaches can enhance trunk muscular endurance and movement proficiency in a relatively short period of time. For gymnasts to realise ongoing adaptations, exercises will need to be progressively overloaded and training prescription will need to be altered. Strength and conditioning practitioners should also be aware that providing technical competency is sufficiently robust, pre-PHV gymnasts may require higher exercise intensities  and longer training durations to facilitate neuromuscular adaptations that improve reactive strength, leg stiffness and fast-SSC function. Importantly, strength and conditioning practitioners must prioritise technical competency at all times and aim to differentiate exercises on an individual basis via regressions and progressions.


Kaposi sarcoma-associated herpes virus (KSHV) is one of the most common causal agents of Kaposi Sarcoma (KS) in individuals with HIV-infections. The virus has gained attention over the past few decades due to its remarkable pathogenic mechanisms. A group of genes, ORF71, ORF72, and ORF73, are expressed as polycistronic mRNAs and the functions of ORF71 and ORF72 in KSHV are already reported in the literature. However, the function of ORF73 has remained a mystery. The aim of this study is to conduct comprehensive exploratory experiments to clarify the role of ORF73 in KSHV pathology and discover markers of AIDS-associated KSHV-induced KS by bioinformatic approaches.We searched for homologues of ORF-73 and attempted to predict protein-protein interactions (PPI) based on GeneCards and UniProtKB, utilizing Position-Specific Iterated BLAST (PSI-BLAST). We applied Gene Ontology (GO) and KEGG pathway analyses to identify highly conserved regions between ORF-73 and p53to help us identify potential markers with predominant hits and interactions in the KEGG pathway associated with host apoptosis and cell arrest. The protein p53 is selected because it is an important tumor suppressor antigen. To identify the potential roles of the candidate markers at the molecular level, we used PSIPRED keeping the conserved domains as the major parameters to predict secondary structures. We based the FUGE interpretation consolidations of the sequence-structure comparisons on distance homology, where the score for the amino acids matching the insertion/deletion (indels) detected were based on structures compared to the FUGE database of structural profiles. We also calculated the compatibility scores of sequence alignments accordingly. Based on the PSI-BLAST homologues, we checked the disordered structures predicted using PSI-Pred and DISO-Pred for developing a hidden Markov model (HMM). We further applied these HMMs models based on the alignment of constructed 3D models between the known structure and the HMM of our sequence. Moreover, stable homology and structurally conserved domains confirmed that ORF-73 maybe an important prognostic marker for AIDS-associated KS.Collectively, similar variants of ORF-73 markers involved in the immune response may interact with targeted host proteins as predicted by our computational analysis. This work also suggests the existence of potential conformational changes that need to be further explored to help elucidate the role of immune signaling during KS towards the development of therapeutic applications. Pre-existing human immunodeficiency virus (HIV) infections affect the immune system increasing the risk for development of Kaposi sarcoma (KS). Since the discovery of Kaposi sarcoma-associated herpesvirus (KSHV), also termed human herpesvirus 8 (HHV8), the tumor development and oncogenesis were associated with co-expression of different genes . KS is aIn fact, KS was named after Dr. Moritz Kaposi, a prominent Hungarian dermatologist, who described KS as an ‘idiopathic pigmented sarcoma of the skin' in 1872 . The evoThe KSHV is closely related to the subfamily Rhadinoviridae (gamma-2-herpesviruses), which is also close to the Herpes virus saimiri (HVS); therefore, similarities between ORFs of KSHV and HVS may influence the pathogenesis of KS . The HVSTypically, the phenotypic features of KS initially appear on the face, legs, or feet as painless red spots but, in severe cases, the lesions also appear in the lungs and digestive tract . KSHV isKSHV infection of endothelial and/or hematopoietic progenitors  alter thThe variability in KS lesions observed in histopathological assays include spindle cell hemangiomas, cutaneous angiosarcomas, vascular leiomyomas, and fibrous histiocytomas . Endothehttps://genecards.weizmann.ac.il/v3/index.php?path=/Search/keyword/kaposi%20sarcoma%20markers/0/20; UniPortKB https://www.uniprot.org/uniprot/?query=kaposi+sarcoma&sort=score; and NCBI https://www.ncbi.nlm.nih.gov/protein/?term=ORF-73%20kaposi%20sarcoma).We used publicly available databases including the National Centre for Biotechnology Information (NCBI), GeneCards  and UniPWe used publicly available internet-based protein search tools and bioinformatics programs with default settings, unless otherwise stated in the text, for the analysis. We tested selected protein sequences to identify conserved domains from NCBI and BLAST algorithms, and we used the PSIPRED program to predict the secondary structure of proteins based on the conserved domain sequences. We further executed a position specific iterative BLAST (PSI-BLAST) search to build a PSSMs (position specific score matrix), which could predict the secondary structure of the input sequences  to prediP value <0.05 as the cut-off criterion.We chose the ORF-73 target effector to perform a Gene Ontology (GO) search, is a hierarchical graph-based annotation system where the terms closer to the root describe more general information while those away from the root provide more specific information about a given GO category and all the GO terms associated with a protein sequence were obtained from the GO database. The KEGG network pathway enrichment analysis by collecting data of related genomes and their pathways associated with diseases  and we shttps://genemania.org/) to analyze interactions associated with KS among the proteins encoded by the DEGs. The two parts of GeneMania algorithm consists of an algorithm based on linear regression to calculate functional association from multiple networks from different data sources; and a label predicting gene function of composite network. We employed keywords such as—ORF73 to determine interacting partners. This was pursued using downstream regulator p53 as an apoptosis marker during pathogenesis in the host. Moreover, the marker protein was used for transient interaction study.We used the online Search Tool for the Retrieval of Interacting Genes (STRING)  and GeneWe immobilized His-tag, GST-tag, or biotin-tag bait proteins to an affinity resin and incubated them with solution expressed proteins as prey proteins. We then captured the bound bait and pulled down the cell lysate flow through. Subsequently, we used mass spectrometry (MS) or Western blots to confirm interactions. Using this technique, we determined interacting protein partners of relevant proteins .Annotations used to search for the KS-associated markers in the UniProtKB database quoted about 137 entries, which we then screened to find those with computationally annotated data. Search engine GeneCards reported about 369 KS markers with a relevance score. We found61 ORF-73 marker homologous hits related to the family of human gamma herpes virus 8 with varied E-values. Out of these, we used only the most identical sequence , AAC57158.1, for our computational analyses. A search for proteins similar to the selected marker ORF-73 resulted in8 protein accessions , and 2 CDS regions (accession numbers AAC57158.1 and AAC55944.1).We looked for conserved domains in the marker protein ORF-73 based on hypothetical domain sequences using literature recapitulation NCBI's Conserve Domain Database (CDD). To identify potential marker roles at the molecular level, we focused on its predicted secondary structure. Therefore, we searched for hypothetical protein having conserved domain and used accession number AAC5744 of gi.1633572 in an NCBI domain search and found only one significant hypothetical conserved domain (PHA03169) with the same accessison number  . To accoPull-down assays serve as a complementary method to further validate the predicted interactions in a quantitative manner towards understanding their dissociation constants and relative bindings of proteins and their direct binding sites. However, this is beyond the scope of this study. We believe the following recommendations should be followed by researchers investigating transient protein interactions: First, determining the protein solubility is essential. If the prey protein is at a too-high concentration, it will not be sufficiently soluble. Second, shortening the time and adjusting buffer conditions of incubation help prevent prey protein degradation. Third, checking the prey protein with beads if bait protein is not bound should be done as a control. Fourth, conducting all assays at a constant temperature of 4 °C should be considered if a variation in Kd is found between repeated experiments.The tumor suppressor antigen p53 depends on specific cellular conditions to induce arrest of cell growth and to control cell division .Our network analysis  showed involvement of LANA and other effector markers in KS conditions and helped elucidate their mechanisms of action , that coMany viral genes are homologous to host cellular genes in KSHV . The PubORF-73 is not the only protein marker implicated in KS pathology, but much about it remains unknown. It is used as a marker for KSHV; especially, its protein folding and motifs are important for the marker assessment observed in the pattern of structural domains in the selected sequence analyzed with PSI-PRED. The pathogenic interactions in the network-based analysis between LANA and the host p53 suggest that LANA was confirmed by STRING and FUGUE tools. The predicted sequence motifs give detailed interactions that are conserved in the subfamilies of the herpesviruses as discussed in detail on the KEGG pathway with notable mechanisms described in the literature . HoweverEscherichia coli  experiments because of our ligand's affinity to capture the strongest interacting proteins . MS idenOur computational studies found that ORF-73 is involved in host apoptosis through p53 signaling pathways and is a key marker associated for Kaposi Sarcoma. This study also identified potential KS-associated genes which are reported to trigger cancer and suggested mechanisms of interaction that may help researcher developing prevention strategies.We retrieved all data from publicly available resources and we required no ethical approvals for dissemination of this purely academic information.PZ, JW, and XZ conceived and designed the study. XW, LJ and XG provided study materials and were responsible for the collection and assembly of data, data analysis, and interpretation. PZ was involved in writing of the manuscript. All authors read and approved the final manuscript.This study was supported by the Seed Fund Program of Shanghai University of Medicine and Health Sciences (Grant No. SFP-18-21-01-002), the General Program of Pudong New Area Health and Family Planning Commission of Shanghai, China (Grant No. PW2016A-7), the National Natural Science Foundation of China (No. 81830052), Construction project of Shanghai Key Laboratory of Molecular Imaging (18DZ2260400), Shanghai Municipal Education Commission , and the Natural Science Foundation of Guangdong Province (2016A030313680).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.