Delivery of drugs to special locations of ocular lesions, while minimizing systemic and local toxic effects, is recognized as a critical challenge in the ophthalmic practice. The special anatomy and physiology barriers within the eyeball entail effective drug delivery systems. Emerging attempts in drug delivery has led to developments in ocular iontophoresis, which acts as a noninvasive technology to enhance drug penetration using a small electric current. This technique offers greater flexibility to deliver desired drug dose in a controlled and tolerable manner. In previous studies, this technique has been testified to deliver antibiotics, corticoid, proteins and other gene drugs into the eye with the potency of treating or alleviating diverse ophthalmological diseases including uveitis, cataract, retinoblastoma, herpes simplex and cytomegalovirus retinitis (CMVR), etc. In this review, we will introduce the recent developments in iontophoresis device. We also summarize the latest progress in coulomb controlled iontophoresis (CCI), hydrogel ionic circuit (HIC) and EyeGate II delivery system (EGDS), as well as overview the potential toxicity of iontophoresis. We will discuss these factors that affect the efficacy of iontophoresis experiments, and focus on the latest progress in its clinical application in the treatment of eye diseases. Therefore, it is permeable to large molecular weight compounds  is one of the most common diseases of the anterior segment Gayton, . There iReactive oxygen species (ROS) scavengers in tears can remove ROS from the surface of the eye. However, if excessive ROS are left on the eye surface, or if tear production is reduced, ROS would impair eye tissue , particularly HSV-1 can affect various components of the human eye, causing herpes simplex keratitis, uveitis, retinitis or endophthalmitis . They found that although the high aqueous solubility of ACV-X prodrugs enhanced passive transcorneal delivery of ACV species, short-duration iontophoresis resulted in significantly greater transcorneal permeation and corneal deposition of ACV species using less concentrated prodrug solutions. As evidenced by the ACV-Gly results, the drug permeation through sclera by iontophoresis showed a significant increase compared with passive diffusion. Due to the high intrinsic permeability of the sclera, the penetration rate of transscleral iontophoresis is much greater than that of transcorneal iontophoresis. Biodistribution studies further confirmed the superiority of short-duration iontophoresis, as the average concentration of ACV species in the eye globes following iontophoresis of ACV-Gly for five minutes was greater than the IC4.5.Noninfectious scleritis is a rare eye disease that is accompanied by redness and severe ocular pain , are therefore of increasing concern  lead to retinal vascular value-added lesions, vitreous hemorrhage, retinitis, and even retinal detachment  can improve DR therapy. Bevacizumab is a humanized IGg1 antibody that binds primarily to VEGF with high affinity to reduce VEGF. But it cannot passively pass through the sclera to reach therapeutic concentrations in the posterior segment. Thus a new study attempted to use iontophoresis to overcome the inability of bevacizumab to pass through the sclera. They concluded that anodic iontophoresis could increase the amount of bevacizumab passing through the sclera, thus demonstrating the promising use of iontophoresis for DR  is the most common primary intraocular malignancy of childhood, and approximately 9000 new cases occur worldwide each year  is a disease of immunocompromised host, often associated with severe vision loss due to retinal atrophy, retinal detachment and/or optic nerve atrophy  is secondary to many fundus diseases, most commonly seen in wet age-related macular degeneration  and pathological myopia (mostly in high myopia (> U0 degrees). In addition, CNV may be complicated by fundus angioid stripes, hereditary macular degeneration, inflammation , trauma, tumor or no obvious cause .By 2010, only two anti-angiogenic drugs have received Food and Drug Administration (FDA) and European Medicines Agency approval for the treatment of neovascular AMD, namely Ozurdex (dexamethasone) and Pegaptanib  would cause retina and choroid burns, hemorrhagic necrosis, edema and infiltrations in the subjects , proliferation of macrophages, and loss of retinal nuclear layer. On the other hand, another study shows that after several weeks of gentamicin iontophoresis through sclera with extremely high current density (765.3 mA/cm2) , corneal epithelial cell edema and endothelial cell count decline occurred within a short period of time (1–10 min) . The lack of funding to promote this technology to medical institutions is currently another reason that preventing this technology from entering the clinic practice. More attention should be paid to this promising technology to enhance its future clinical utilization.


This cross-sectional study investigates changes in the workforce providing contraception and abortion services from before to during the COVID-19 pandemic. These include COVID-19 and policies penalizing abortion care, which culminated in the 2022 Dobbs v Jackson decision. Using a national-level claims data set, we examined changes in the contraception and abortion workforce from 2019 through 2021.The clinical workforce providing contraception and abortion care is critical for access. This workforce includes women’s health, primary care, and other clinicians.STROBE reporting guideline.This cross-sectional study was approved by the George Washington University institutional review board, which waived informed consent because there was no more than minimal risk to participants. This study is reported following the We used 2019 to 2021 IQVIA preadjudicated medical claims derived from clinician offices and insurance clearinghouses, including clinician month-level counts of services for in-person contraception , contraceptive implants, and injectable contraception), abortion , or both. Outcome variables were counts of contraception clinicians and abortion clinicians by year and service type and monthly volume changes in services. During January 2019 to December 2021, 141 837 clinicians provided at least 1 in-person contraception or abortion service; 5804 clinicians provided both services. Data analyses occurred from June 1 to July 10, 2022, using Stata statistical software version 17.0 (StataCorp).The number of physicians decreased in 2020 for all services except medication abortion, and the number of physicians providing contraception was not reestablished to prepandemic levels in 2021. By contrast, the number of advanced practice clinicians stayed nearly identical from 2019 to 2020 and increased above prepandemic levels in 2021 .Contraception services decreased substantially from March through May 2020 but were restored to approximately prepandemic levels later in 2020 and through 2021 . Abortio2In this cross-sectional study, we found that the overall number of clinicians providing contraception and abortion services decreased in 2020 and was restored in 2021. However, the number of physicians providing contraception services did not reach prepandemic levels, while the number of advanced practice clinicians providing contraception services continued to increase, mirroring the increasing numbers of advanced practice clinicians in primary care and other settings.4 While these physicians may reenter the workforce, our findings suggest there is an ongoing loss of physicians providing contraception services, which is concerning. Access requires that primary care clinicians offer the full scope of care, including family planning services. Targeted investments in women’s health and primary care clinicians and state-level expanded scope of practice policies may strengthen this segment of the workforce.COVID-19 placed incredible strain on the health workforce, with reports of practice closures and physicians leaving practice.5 In 2020, many individuals reported fertility plans for fewer children or later pregnancies, suggesting that demand for contraceptive services remained even as services were unavailable.6We also found that in-person contraception services decreased in the early COVID-19 period (March-May 2020) but were restored later in 2020, However, procedural abortion services decreased over the 3-year period, and medication abortion services increased, consistent with other studies.1 Additionally, this study did not capture outcomes associated with the 2022 Supreme Court decision overturning Roe v. Wade, a watershed event that will shape the provision of abortion and contraception for the foreseeable future. However, we found losses in this workforce even before 2022. It will be critical to continue to track this workforce in the coming years given that access to care intrinsically relies on the workforce.Although IQVIA medical claims contain data on 94% of physicians registered with the American Medical Association, this study provided a partial view of the contraception and abortion workforce owing to data limitations noted elsewhere.


EGFR) oncogene addicted tumor. In these patients, despite the clear benefits of tyrosine kinase inhibitors therapy, the development of acquired resistance and progressive disease is inevitable in most cases and liquid biopsy is important for molecular characterization at resistance and, being non-invasive, may be useful for disease monitoring. In this review, the authors will focus on the applications of liquid biopsy in EGFR-mutated non small cells lung cancer at diagnosis, during treatment and at progression, describing available data and possible future scenarios.Liquid biopsy has emerged as a minimally invasive alternative to tumor tissue analysis for the management of lung cancer patients, especially for epidermal growth factor receptor ( Liquid biopsy consists in the analysis of tumor circulating materials derived from body fluids. Since a plethora of anticancer drugs are currently accessible for lung cancer treatments, an iterative assessment of tumor’s biomarkers is the ideal approach to define the best sequence of therapies for each patient. In this scenario, liquid biopsy can be used as a non-invasive method to detect any targetable genetic mutations and to select a corresponding targeted therapy, to monitor response to treatment and to identify eventual genomic alterations occurred at resistance. Moreover, liquid biopsy provides information on the genomic profile of the whole tumor landscape, overcoming spatial and temporal heterogeneity .From body fluids samples of metastatic cancer patients, the circulating free DNA (cfDNA) is usually found in small quantities. Only a small portion of cfDNA derives specifically from tumor cells, defined as circulating tumor DNA (ctDNA), and can be analyzed by next-generation sequencing (NGS) . NGS is Liquid biopsy is extremely relevant in lung cancer patients, as the tumor is often difficult to reach with surgical or radiological biopsies, which are anyway invasive and potentially risky procedures. In fact, almost three-fourths of lung cancers patients with advanced disease are diagnosed via cytology samples because the acquisition of histologic specimens is not possible, resulting in a 30% of cases in which the sample is not sufficient for biomarker evaluation, either at diagnosis or at disease progression .EGFR) mutations occur in almost 10–12% of cases and tyrosine kinase inhibitors (TKIs) are the standard front-line therapy  NSCLC, including genomic alterations that are detectable by liquid biopsy as described below.Some data of mechanisms of resistance to first line osimertinib are already available as is now the standard of care for Otherwise, in patients with T790M-NSCLC a tumor re-biopsy is recommended to define the tissue T790M status and, if confirmed as negative, platinum doublet chemotherapy is the standard second-line treatment .MET amplification (estimated to occur in 5% to 20% of patients) or HER2 amplification (in up to 8% of patients), activation of other pathways like AXL, Hedgehog and downstream signals  .HER2 amplification-mediated resistance in EGFR-T790M+ NSCLC cell lines [EGFR-mutant NSCLC treated with a sequence of first-generation EGFR-TKI, second-generation EGFR-TKI afatinib and osimertinib [Moreover, other data are available from preclinical studies. The combination of osimertinib and trastuzumab-emtansine (T-DM1) has been shown to overcome ll lines . Also, omertinib .HER2 deletion causing resistance to osimertinib in an EGFR-T790M+ patient. HER2 exon 16 skipping mutation has only previously been described in breast cancer, in which it is reported as an oncoprotein that activates Src kinase signaling. The mutated tumor cells resulted to be resistant to Src inhibition with or without osimertinib, but the combination therapy with osimertinib and the pan-HER2 TKI afatinib may be effective. As the mutation was already present before treatment, this case highlights the relevance of obtaining a genomic characterization to determine preexisting alterations which may have both prognostic and predictive value for therapeutic implications; similarly, this study also demonstrates that repeating molecular analyses at multiple time points is useful to discover new potentially actionable molecular targets [Hsu et al. , recentl targets .MYCL1, SOX2, and SOX4. In addition, TP53 mutations were found to be at low levels before EGFR-TKI initiation in patients baseline plasma samples. Being TP53 mutation universally present in all SCLC, the pre-existing clone of TP53 mutant cells may growth selectively under the pressure of EGFR-TKIs treatment and could explain the SCLC-like resistance mechanism. The levels of TP53 mutant cells in plasma were also higher with disease progression and lower when patient experienced clinical response during SCLC-directed chemotherapy [Another mechanism of resistance to late generation TKIs treatment is reported to be the transformation to SCLC, known to be difficult to assess through liquid biopsy . Neverthotherapy , 36.MET amplification (15%), followed by EGFR C797S mutation (7%); other mechanisms described were HER2 amplification (2%), PIK3CA and RAS mutations (2–7%) [As in the AURA3 trial, in the phase III FLAURA study, paired plasma samples were collected at baseline and at PD and/or treatment discontinuation and were analyzed using NGS to determine alteration that may have caused acquired resistance to first-line osimertinib. Preliminary data revealed that in the osimertinib arm, there was no evidence of acquired EGFR T790M, as expected knowing the specificity of osimertinib for T790M, and the most frequent acquired resistance mechanism detected was s (2–7%)   is a phase II, open-label, single-arm trial designed to evaluate the efficacy, safety and resistance mechanisms to osimertinib when administered as first-line therapy in patients with locally advanced or metastatic We are also expecting the results of the phase II ORCHARD trial (NCT03944772) that aims to investigate molecular-guided treatment options as second line after progression on osimertinib by evaluating potential targetable acquired resistance mechanisms. Patients will receive study treatment according to the mutation encountered: osimertinib plus gefitinib, osimertinib plus necitumumab,osimertinib plus savolitinib, platinum-based doublet plus durvalumab.As stated above, liquid biopsy can be utilized to dynamically detect resistance mutations in course of treatment with EGFR inhibitors and, when the tumor tissue biopsy is not possible, ctDNA analysis is the only available method for the identification of patients that can benefit from a TKI therapy.However, an important limitation is the unsuccessful concordance between the mutational status of cfDNA and tumor DNA, maybe due to the low quantity of ctDNA that makes difficult the evaluation of the heterogeneity of tumor mutational landscape. To overcome this issue, other body fluids in sites adjacent to metastases have been studied as an alternative and more reliable source of ctDNA. In fact, higher levels of cfDNA were extracted from other body fluids such as cerebrospinal fluid, pleural effusion, and ascites, resulting in a valuable method for detecting tumor biomarkers .In addition to the cfDNA, circulating tumor cells (CTCs), miRNAs and exosomes are other tumor-derived material that could be reliable surrogates to investigate tumor biology. In a pre-clinical setting these evaluations revealed promising results for their possible application in screening programs or as prognostic/predictive biomarkers and their applications will probably shift to the clinical scenario in the near future.CTCs reflect tumor properties providing genomic, transcriptomic, and proteomic information on the tumor and, in addiction, may also be used to create preclinical models such as organoid cultures and murine models –40. TumoLiquid biopsy already implemented clinical management of patients at diagnosis, during treatment and at progression offering a biomarker-oriented therapeutic option. Before starting treatment, liquid biopsy can be essential to investigate not only target mutations but also secondary mutations that can lead to resistance by clonal selection. During treatment, repeating serial liquid biopsies could have a role in monitoring disease evolution and anticipating a potential progression. At progression, liquid biopsy may identify mechanisms of resistance to select an appropriate treatment according to the therapy-dependent clonal selection, considering also that in the later-line osimertinib study, 36% of patients developed resistance through a targetable oncogene alteration. Nevertheless, no clear mechanism of resistance is established in approximately 40% of patients treated with second line osimertinib and in 50% of patients treated in first-line setting. Therefore, furthers trials to identify these unknown mechanisms are needed.


Ribosome profiling quantifies translation genome-wide by sequencing ribosome-protected fragments, or footprints. Its single-codon resolution allows identification of translation regulation, such as ribosome stalls or pauses, on individual genes. However, enzyme preferences during library preparation lead to pervasive sequence artifacts that obscure translation dynamics. Widespread over- and under-representation of ribosome footprints can dominate local footprint densities and skew estimates of elongation rates by up to five fold. To address these biases and uncover true patterns of translation, we present choros, a computational method that models ribosome footprint distributions to provide bias-corrected footprint counts. choros uses negative binomial regression to accurately estimate two sets of parameters: (i) biological contributions from codon-specific translation elongation rates; and (ii) technical contributions from nuclease digestion and ligation efficiencies. We use these parameter estimates to generate bias correction factors that eliminate sequence artifacts. Applying choros to multiple ribosome profiling datasets, we are able to accurately quantify and attenuate ligation biases to provide more faithful measurements of ribosome distribution. We show that a pattern interpreted as pervasive ribosome pausing near the beginning of coding regions is likely to arise from technical biases. Incorporating choros into standard analysis pipelines will improve biological discovery from measurements of translation. Quantification of gene expression has enabled insights into many aspects of biology, often relying on mRNA abundances as a proxy for protein levels , 2. RiboThe quantitative aspect of ribosome profiling has proved a rich source of information from which to study translation dynamics. Each footprint recovered in a ribosome profiling dataset reflects the position of an individual ribosome on a transcript, resulting in precise codon-level resolution of the distribution of ribosomes across a transcript. Ribosomes are distributed unevenly along individual transcripts, reflecting biologically significant phenomena such as sites of ribosome pausing or stalling and inherent differences in elongation rate over different sequences. Per-position ribosome abundances can be used to infer translation elongation rates: the slower the elongation rate, the longer the dwell time and the more likely a ribosome is observed at that codon position. Analyses of ribosome profiling datasets have revealed that the largest driver of elongation rate is the identity of the codon being decoded in the aminoacyl (A) site of the ribosome, with relationships between elongation rate and tRNA availability, wobble base-pairing, and amino acid properties –10. ElonHowever, experimental artifacts can distort estimates of translation rates, models of translation dynamics, and signals of ribosome pausing. A particularly strong distortion arises from sequence-based biases introduced during certain steps of library preparation protocols, namely, ligation of an adapter to the 3′ end of the footprint and circularization of the reverse transcription product in order to ligate an adapter to the 5′ end . These bSequence preferences have been previously identified in other types of RNA sequencing experiments; however, in these cases the reads are summed across a full transcript or a wide window of sequence, and biases are likely to average out across the many different sequence contexts in one transcript –22. WhenTo this end, we present choros, a computational method that estimates biological and technical contributions to ribosome distributions and generates sequence bias-corrected footprint counts. We validate choros on data simulated with realistic sequence biases and demonstrate the method’s ability to capture underlying parameters. Applied to experimental ribosomal profiling datasets, choros correctly attenuates ligation biases and provides accurate estimates of relative ligation probabilities and codon-level ribosome occupancies. The extensible regression framework also enables detection of differential codon-level translation between experimental conditions. Using choros, we show that bias correction removes large artifacts that have been interpreted as biological signals and enables detection of biological phenomena such as ribosome collisions. choros is implemented as an open-source R package that can be incorporated into standard ribosome profiling analysis pipelines to provide accurate ribosome footprint counts for downstream analysis and biological discovery.https://github.com/lareaulab/choros. simRiboSeq is an R package for simulating ribosome profiling data, available on GitHub at https://github.com/lareaulab/simRiboSeq. Code to regenerate figures in this work are available on GitHub at https://github.com/lareaulab/choros_paper.Our software, choros, is provided as an R package for correcting sequence biases in ribosome profiling datasets and is available on GitHub at Our method begins with a model of ribosome profiling data that is based on the physical processes that generate ribosome footprints: (i) a biological component that governs a true distribution of ribosomes, from which (ii) experimental protocols sample with bias to generate a sequencing library . OverallWe fit a generalized linear model (GLM) to estimate the parameters underlying this data-generating process . The modWe include interaction terms for digestion length and bias sequence at each end of the footprint to capture possible sequence preferences of nuclease digestion as well as effects of non-templated nucleotide additions during reverse transcription . The regTo reverse the effects of biased footprint recovery, we generate a correction factor for each individual footprint based on the parameter estimates learned in the regression fit :(6)coglm.nb from the MASS package and performing bias correction, choros outputs a table of raw and corrected read counts, aggregated by transcript, codon position, and 5′ and 3′ digestion lengths.choros has four major modules: data processing (including A-site assignment and bias sequence determination), regression fitting, bias correction, and bias evaluation . The inpFootprints were assigned to transcript codon positions using an A-site assignment process previously described in Tunney et al. . In briechoros is flexibly designed to fit the regression model on a user-supplied list of transcripts and codon positions. While the technical biases that arise during library preparation should be universal to footprints from all transcripts, sparsity in read coverage may lead to unreliable parameter estimates. Unless otherwise described, all regression models in this work were fit to a random set of 250 of the top 500 transcripts according to footprint coverage (mean footprint abundance per codon position) after excluding transcripts that had fewer than 5 mean reads per codon position and fewer than 100 codon positions with mapped footprints. In addition, the first and last 20 codon positions of transcripts were excluded from the regression fit to avoid the effects of translation initiation and termination.Bias correction was performed as described above . Biases To test choros, we developed an R package simRiboSeq to simulate ribosome profiling datasets with realistic parameters for individual steps in the data-generating process . Per-traValues for simulation probabilities were adapted from experimental datasets. Per-transcript probabilities were generated from per-transcript ribosome abundances extrapolated from a loess regression of per-transcript ribosome abundances on transcript lengths fit to data from Weinberg et al. with additional Poisson noise . Per-codThe experimental datasets analyzed in this work are listed in https://github.com/ncbi/sra-tools). Trimming of 3′ adapters was performed with cutadapt, and trimmed reads longer than 20 nt were retained with options ‘--trimmed-only -m 20’ . Wh. Wh29]. In keeping with prior results , 29, we We then confirmed the ability of choros to attenuate these biases. After bias correction, the position importance plots showed a decrease in signal at the ribosome boundary positions and an increase in signal at the ribosome P and A sites . The ploNext, to see if bias correction enabled more accurate measurement of footprint distribution along individual transcripts, we evaluated the concordance in footprint distribution between the fixed-adapter and random-adapter datasets, before versus after bias correction. Footprints were aggregated by A-site codon position, and footprint counts were compared between the two datasets. The two datasets were indeed more consistent after bias correction, with the overall Spearman correlation between datasets increasing from 0.73 to 0.84. The improvement from bias correction varied by transcript, with a strong dependence on transcript coverage . While tSpikes in footprint counts—whether caused by ligation biases or genuine translational stalls or pauses—can be visible in ribosome profiling data on individual genes, but these signals are expected to be averaged out in aggregate data such as metagene plots showing the overall distribution of ribosomes over coding regions. However, some pervasive, position-specific spikes do appear in aggregate data. The overabundance of footprints on the sixth codon of reading frames, visible in metagene plots, has been interpreted as an indicator of a pause while the ribosome commits to elongation . We reasWe observed a strong peak of ribosome footprints at the sixth codon in all of the analyzed datasets that were generated with CircLigase protocols . After bchoros fits a generalized linear model to model the biological and technical processes underlying ribosome profiling data. The extensibility of this regression framework allows the inclusion of additional terms. One such parameter is the detection of differential codon-level ribosome occupancy under two experimental conditions . The decWe assessed this approach on a dataset where yeast cells were treated with 3-amino-1,2,4-triazole (3-AT), a known inhibitor of histidine biosynthesis . When fiRibosome collisions are widespread throughout the transcriptome, particularly on highly translated transcripts loaded with multiple ribosomes , 43–45. Collisions of a 5′ elongating ribosome into a 3′ paused ribosome can result in longer protected fragments, or disome footprints, that can be recovered by ribosome profiling using a broader size selection . AnalyseWe applied this approach to a dataset of monosome- and disome-protected fragments . PositioTo examine whether bias correction with choros was able to remove sequence artifacts in disome counts, we generated position importance plots across the span of a disome footprint. We observed an increase in signal at the positions corresponding to the disome fragment boundaries in raw counts, indicative of sequence artifacts as also seen in monosome data .This sigRibosome profiling informs our knowledge of gene expression by quantifying translation of mRNAs. Beyond this, it also provides a codon-resolution snapshot of ribosome distribution across transcripts, allowing inference of the decoding time of different codons and providing insight into molecular mechanisms of translation. This codon-level information has led to advances in synthetic mRNA design and discovery of amino acid limitations and tRNA alterations in cancers , 46–48. Sequence-dependent biases are pervasive across ribosome profiling datasets, even when careful attention is paid to the library preparation protocol. This arises from the necessary biochemical manipulation of ribosome-protected footprints to make them amenable to high throughput sequencing. For instance, most library preparation approaches require ligation of sequencing adapters onto the ends of the footprints. Ligases generally have sequence preferences, likely for both the 5′ and 3′ entities to be ligated, making these biases unlikely to be completely avoidable. The resulting over- and under-representation of certain footprints leads to distorted ribosome distributions, which can be detected as artifactual signals showing an influence of several nucleotides at either end of footprints on footprint recovery , 15, 16.Our method, choros, directly estimates ligase preferences and generates correction factors to produce footprint counts that are free from these sequence-dependent biases. Careful statistical analysis, use of simulated data, and comparisons with our experimental data on ligation preferences confirm the validity of our model. choros was able to reverse the effects of ligation biases across many experimental datasets, including disome footprints, and also simultaneously produced parameter estimates that were interpretable and comparatively free of sequence biases. Using our correction method, we showed that a signal often interpreted as a biological phenomenon was potentially the result of biased footprint recovery. Our method will therefore allow more accurate detection of biologically relevant ribosome pauses: interactions with regulatory proteins; intrinsic pauses due to tRNA abundance, peptide bond formation, or nascent chain interactions; collisions between ribosomes; and other as-yet-undiscovered phenomena.The quantitative model stems from a conceptual data-generating process that can be generalized to multiple library preparation protocols, including dual ligation and template jumping approaches. However, even with choros-corrected footprint counts, interpretation of codon-level data from ribosome profiling presents challenges. Ribosome profiling datasets can also suffer from other types of biases. For instance, translation elongation inhibitors such as cycloheximide, used to stabilize ribosomes on mRNAs, can lead to ribosome accumulation on certain codons , 52.The The sparsity of footprint data on all but the most abundant or highly translated transcripts presents a known limitation in interpreting ribosome profiling data. This limitation extends to our bias correction method. In fact, when no footprints are recovered at a position, whether because of sparse ribosome density or poor footprint recover, bias correction is not possible; choros adjusts footprint counts in a multiplicative manner that cannot add footprint counts to positions where none were originally observed. In this analysis, we have used only the most highly translated transcripts to learn the biological and technical parameters governing ribosome distribution. We emphasize that the combination of sparse data and sequence biases renders many transcripts unsuitable for detailed individual analysis by ribosome profiling, while aggregate analyses of patterns are highly reliable. As such, we consider choros most appropriate for analyzing translation of abundant transcripts in high-coverage datasets. To mitigate the impact of missing data, future work could explore the addition of pseudocounts in order to restore counts to positions where biased recovery led to no observations, and to increase the reliability of ribosome distributions on sparse transcripts.In summary, choros provides a much-needed solution to the problem of sequence-based biases in ribosome profiling data and should be incorporated into standard analysis pipelines to provide faithful codon-resolution measurements of translation and enable biological discovery.Supplement 1


Understanding the specific molecular mechanisms of basal docking is an area ripe for further exploration that will shed light on complex morphogenetic events that sculpt developing organisms and on the cellular mechanisms that can go awry during diseases involving the formation of cysts, fistulas, atresias, and metastases.Epithelial tissues form continuous barriers to protect against external environments. Within these tissues, epithelial cells build environment-facing apical membranes, junction complexes that anchor neighbors together, and basolateral surfaces that face other cells. Critically, to form a continuous apical barrier, neighboring epithelial cells must align their apico-basolateral axes to create global polarity along the entire tissue. Here, we will review mechanisms of global tissue-level polarity establishment, with a focus on how neighboring epithelial cells of different origins align their apical surfaces. Epithelial cells with different developmental origins and/or that polarize at different times and places must align their respective apico-basolateral axes. Connecting different epithelial tissues into continuous sheets or tubes, termed epithelial fusion, has been most extensively studied in cases where neighboring cells initially dock at an apical-to-apical interface. However, epithelial cells can also meet basal-to-basal, posing several challenges for apical continuity. Pre-existing basement membrane between the tissues must be remodeled and/or removed, the cells involved in docking are specialized, and new cell-cell adhesions are formed. Each of these challenges can involve changes to apico-basolateral polarity of epithelial cells. This minireview highlights several  The fundamental function of epithelial cells is to encase internal tissues to protect them from external challenges while allowing for regulated interactions with the environment. A defining feature of metazoan epithelial tissues is the polarization of cells so that the plasma membrane is divided into an apical surface that faces the “external” environment  and a basolateral surface that faces internal tissues [reviewed in ]. Cell-cAs an example, consider the formation of the most common basic body plan present in bilaterian animals , a “throC. elegans intestine depends upon the presence of the conserved protein scaffold PAR-3, while the C. elegans epidermis can polarize in the absence of PAR-3 (Connecting different epithelial tissues together requires the coordination of apico-basolateral polarity across multiple scales . At the of PAR-3 . During of PAR-3  and instof PAR-3 . Regardlof PAR-3 .in vivo work has found that local and global polarity establishment are temporally and genetically separable during the development of the C. elegans intestine (Apico-basolateral polarization of an epithelial tissue is only functional if neighboring epithelial cells all orient uniformly to form a continuous barrier. At a local scale within a tissue, epithelial cells coordinate with immediate neighbors to align apico-basolateral axes. At a global scale across a tissue and even across entire organ systems such as a through gut digestive tract, this local polarity must be coordinated into a global apico-basolateral polarity that is consistent across all connected epithelial cells, regardless of their origin or their polarity establishment mechanisms . Recent ntestine . In the ntestine . Global ntestine , as showntestine  and zebrntestine . The exivia neighbor exchange can bring formerly separated epithelial cells next to one another, these processes typically occur within a field of similar cell types with a common developmental origin. Cells of different types and/or origins become new neighbors by various different ways, some of which are diagrammed in Complex organ formation requires the connection of cells of different types and origins. While the processes of cell division, cell death, cell extrusion, and lateral intercalation Drosophila renal tubes, principal cells (Xenopus epidermis, the nascent apical surface is constrained by the geometry of overlying cells and forms an apical surface only at vertices (+ vesicles (Cells can radially intercalate into an existing epithelium  to diveral cells , blue foal cells , pink inal cells . During al cells . In addial cells . In othevertices  through vesicles  and throvesicles .via a process recently termed neolamination (Drosophila egg chamber, border cells (Instead of joining tissue types through the intercalation of individual cells, a migrating cluster of cells can join an epithelium mination  (Figure mination . Neolamimination . Both cemination , during er cells , pink deer cells , blue toer cells . While ter cells . Speculaer cells  may undeer cells  and onlyer cells , which mDrosophila dorsal closure (Epithelial tissues can also join on much larger scales when large sheets of cells fuse, fold, and sort to correctly shape organs. Many classical models of epithelial-to-epithelial connection involve epithelial sheets folding to create internal tubes or are cases of epithelial fusion and sorting . In thes closure  in which closure . Neighbo closure . In the  closure  or by in closure .While apical-to-apical epithelial fusion events are well-studied, recent investigations of development have revealed instances of basal-to-basal epithelial fusion , includiXenopus mouth formation, Hedgehog signaling and Wnt antagonists induce basement membrane breakdown between the ectoderm and endoderm (A key challenge in connecting epithelial tissues that initially meet basal-to-basal and reorient into a continuous epithelium is that joining cells must often contend with at least one layer of basement membrane that separates the tissues. During basal docking, the basement membrane presents a physical barrier between the tissues, and regulated basement membrane breakdown is a key general feature of basal docking between epithelial tissues. In mouse kidney development, loss of the basement membrane between the distal renal vesicle cells and the collecting duct is an early step in connecting these two epithelial tissues  Figure . In the endoderm . When baendoderm .C. elegans uterine anchor cell, which invades from the uterus into the vulval epithelium during larval development  (Figureiewed in . Netrin iewed in  polarizeiewed in . The poliewed in . The proiewed in , removinlubility , and forlubility . In factlubility .via basement membrane breakdown and protrusion formation in this system, the surrounding uterine cells and invaginating vulval cells also play key roles in completing this morphogenetic event. Surrounding cells help enlarge and then stabilize the breach in the basement membrane created by the invasion process. Uterine cells limit trafficking of a basement membrane receptor to the cell surface to promote basement membrane sliding away from the breach, and the underlying invaginating vulval cells stabilize the edge of the breach via integrin-based adhesions (in vivo imaging techniques combined with a tissue-specific conditional allele have shown that EGFR signaling within the anchor cell is required to stabilize and align the nascent adherens junctions that connect the anchor cell to the vulF cells (Although the anchor cell is the major specialized cell that promotes tissue fusion dhesions . As the dhesions  to promodhesions . How adhlF cells . Once thlF cells  to form C. elegans pharyngeal cells no longer polarize towards a central common midline to form one lumen but rather invert their apico-basolateral polarity, leading to the formation of two lumens (Drosophila midgut, a specific laminin subunit secreted by the underlying mesoderm is required for the re-polarization of migrating endodermal cells (Drosophila midgut, intercalating intestinal stem cells do not require any of the known canonical apical, junctional, or basolateral polarity complexes to establish apico-basolateral polarity, but do rely upon basal integrin signaling to polarize as these cells differentiate into enterocytes and join the intestinal epithelium (In addition to presenting a physical challenge to basal docking, laminin-rich basement membranes can function in some systems and cell types as a cue to orient, maintain, and even establish apico-basolateral polarity . For exao lumens . In deveal cells . In the ithelium .C. elegans digestive tract development, placement of the basement membrane is important in the connection of the polarized intestine to the polarizing pharyngeal valve cells (While these examples show evidence for the role of basement membrane in establishing and maintaining apico-basolateral polarity within individual developing epithelia, other examples show that controlling the initial presence or absence of the basement membrane is crucial for globally aligning fusing epithelial tissues. During ve cells . At the ve cells , ectopicve cells . Mutatiove cells . Finallyve cells ] follow ve cells , indicatThe specialized cells that contact tissue ends and connect at basal surfaces during epithelial fusion are different from the rest of the cells within a tissue (dark red cells, Kidney morphogenesis is a particularly well-studied example in which a specialized subset of cells facilitates epithelial fusion. Kidney morphogenesis requires the fusion between two tissues, the ureteric collecting duct epithelium and the renal vesicle, a cyst which coalesces as cells undergo an MET . The disAlthough the identity of these specialized, invasive cells in normal renal development is clear, the cells and/or cues that promote invasive and then repolarization behavior of the distal renal vesicle cells remain unknown . Severalin vivo context with tissue-specific approaches. Consequently, remarkably little is known about the specific adhesion molecules that mediate different in vivo docking interactions. Stable, specific adhesion between two different epithelial tissues is a necessary step in establishing a continuous epithelium with aligned apico-basolateral polarity (Drosophila germband epithelium (To form continuous epithelia of the complicated organs found in metazoans, sheets and tubes must fuse and connect. As mesenchymal or other surrounding tissues are likely crucial to promoting correct epithelial connections , these ppolarity . One intpolarity , particupolarity  and the ithelium  are bothvia differential cadherin-based adhesion (In returning to the example of the through gut digestive tract, foregut cells must correctly adhere and align with midgut cells on one side and with epidermal cells on the other side. These correct connections across neighbors of different tissues are crucial for organ function and organism survival. Specific epithelial fusion events between different tissues may arise through preferential stabilization of certain cell-cell contacts, just as specific cells sort into separate tissues adhesion , Ephrin-adhesion , and capadhesion . Determi


Coronary atherosclerosis is a complex, multistep process that may lead to critical complications upon progression, revolving around plaque disruption through either rupture or erosion. Several high-risk features are associated with plaque vulnerability and may add incremental prognostic information. Although invasive imaging modalities such as optical coherence tomography or intravascular ultrasound are considered to be the gold standard in the assessment of vulnerable coronary atherosclerotic plaques (VCAPs), contemporary evidence suggests a potential role for non-invasive methods in this context. Biomarkers associated with deleterious pathophysiologic pathways, including inflammation and extracellular matrix degradation, have been correlated with VCAP characteristics and adverse prognosis. However, coronary computed tomography (CT) angiography has been the most extensively investigated technique, significantly correlating with invasive method-derived VCAP features. The estimation of perivascular fat attenuation as well as radiomic-based approaches represent additional concepts that may add incremental information. Cardiac magnetic resonance imaging (MRI) has also been evaluated in clinical studies, with promising results through the various image sequences that have been tested. As far as nuclear cardiology is concerned, the implementation of positron emission tomography in the VCAP assessment currently faces several limitations with the myocardial uptake of the radiotracer in cases of fluorodeoxyglucose use, as well as with motion correction. Moreover, the search for the ideal radiotracer and the most adequate combination (CT or MRI) is still ongoing. With a look to the future, the possible combination of imaging and circulating inflammatory and extracellular matrix degradation biomarkers in diagnostic and prognostic algorithms may represent the essential next step for the assessment of high-risk individuals. As a leading cause of morbidity and mortality, atherosclerotic cardiovascular diseases are constantly at the forefront of scientific research to further unveil their pathophysiologic basis and to develop appropriate diagnostic and therapeutic approaches. Undoubtedly, the development of a plaque is the critical complication of atherosclerosis and the investigation of its features may guide a tailored management of patients with atherosclerotic cardiovascular diseases. In the context of coronary artery disease (CAD), significant progress has been made towards the assessment of coronary atherosclerotic plaques, since several characteristics have been identified as high-risk for rupture or erosion, leading to the development of acute coronary syndromes (ACS). According to these properties, the plaques which are prone to rupture or erosion may be defined as vulnerable plaques. Although invasive imaging methods remain the gold-standard in the evaluation of vulnerable plaques, non-invasive modalities may be critical in the early detection of those abnormalities. In this review article, we summarize the latest advances in the non-invasive assessment of vulnerable coronary atherosclerotic plaques (VCAP).The process of atherosclerosis is complex, consisting of multiple steps. The invasion of low-density lipoprotein (LDL) molecules in the subendothelial space through the aid of extracellular matrix proteoglycans and the ensuing LDL oxidation consist of the pivotal initial step . EndotheFollowing their establishment, coronary atherosclerotic plaques progress through the continuous accumulation of lipids, the proliferation of VSMCs, and the decreased synthesis and increased degradation of collagen . The thip < 0.001), together with plaque burden exceeding 70% and minimal luminal area ≤4 mm2  together with a history of CAD was independently associated with incident coronary events  . Utilizi< 0.001) . The pre< 0.001) . In anot< 0.001) . Specifi< 0.001) . Compare< 0.001) . Among t< 0.001) . Intensi< 0.001) , thus prAs far as contrast-enhanced cMRI is concerned, early contrast enhancement of a coronary plaque may also be a sign of vulnerability, as it is more frequently encountered in cases of unstable angina pectoris compared to patients with stable CAD . MoreoveThe increasing frequency in the use of nuclear imaging techniques in cardiovascular diseases has not spared the assessment of VCAP. Initial studies were conducted in cancer patients, with 18-fluorodeoxyglucose (FDG) PET/CT detecting significant correlations of target-to-background ratio (TBR) in the region of the left anterior descending artery with cardiovascular risk factors, pericardial fat volume, and calcified plaque burden . However18-sodium fluoride (NaF) is an alternative tracer that has been used, and is indicative of calcification and macrophage activity. As there is limited myocardial uptake through the use of this tracer, motion correction techniques have been additionally applied to enhance coronary artery plaque visualization, with an encouraging 46% reduction of image noise being achieved . Triple-Using tracers that target specific plaque components is also being investigated. Interest has been shown towards (68)Ga-DOTATATE, a tracer that binds to somatostatin receptor 2 that is expressed in macrophages. In patients with neuroendocrine tumors, the use of this tracer in PET/CT demonstrated significantly higher TBR in atherosclerotic plaques compared to normal coronary arteries . ThroughEven though PET/CT has received most of the attention, hybrid PET/MRI imaging methods may be considered as an option, even though their use has been mostly experimental to this point, aiming at improving image quality . In the To conclude, the presented evidence concerning the progress of non-invasive modalities in the assessment of VCAPs indicates the extensive knowledge we have attained with regard to the process of coronary atherosclerosis. Identification of such adverse plaque characteristics may thus represent an appealing option in the holistic management of patients with CAD by providing incremental prognostic information and tailoring the therapeutic approach. Published studies have shown a good correlation with invasive methods or even plaque histology. On top of that, abnormalities in circulating inflammatory and extracellular matrix degradation biomarkers could indicate an additional risk marker. However, the lack of large-scale, multicenter randomized clinical trials and registries is a deterring factor for the widespread implementation of these modalities in everyday clinical practice. Moreover, uncertainties remain regarding the optimal imaging method of choice, with most data stemming from CCTA studies. Limited evidence is available from nuclear imaging studies, which may also face certain limitations concerning myocardial uptake and motion correction that ought to be resolved. Therefore, upcoming studies should be adequately designed to provide the needed answers in the existing evidence gaps and prove the incremental value of the non-invasive, multimodality assessment of VCAPs.


The pro-inflammatory responses and proapoptotic signaling pathways predominantly mediated through TNFR1, while neuroprotective and anti-apoptotic signals induced by TNFR2. In this review, we attempt to discuss the involvement of TNF receptors (TNFRs) and their signaling pathway in ocular tissues with focus on RGC and glial cells in glaucoma. This review also outlines the potential application TNFRs agonist and/or antagonists as neuroprotective strategy from a therapeutic standpoint. Taken together, a better understanding of the function of TNFRs may lead to the development of a treatment for glaucoma.Glaucoma is an irreversible sight-threatening disorder primarily due to elevated intraocular pressure (IOP), leading to retinal ganglion cell (RGC) death by apoptosis with subsequent loss of optic nerve fibers. A considerable amount of empirical evidence has shown the significant association between tumor necrosis factor cytokine  and glaucoma; however, the exact role of TNF in glaucoma progression remains unclear. Total inhibition of TNF against its receptors can cause side effects, although this is not the case when using selective inhibitors. In addition, TNF exerts its antithetic roles  Glaucoma, an ocular neurodegenerative disease, is a leading cause of irreversible blindness that has affected over 70 million people worldwide . The halThe axonal damage caused by injury to the optic nerve is thought to be the first stage of insult in glaucoma. However, direct impact on the optic nerve may ultimately result in RGCs death, which is exacerbated by the production of inflammatory cytokines from resident innate immune cells (glia) and the complement system in retina . Meanwhivia TNF receptor I (TNFR1) and TNF receptor II (TNFR2). Various cellular responses can elicit TNF expressions through these receptors. It depends on many factors, including the metabolic state and the adaptor proteins present in the cells  , 10. Sinhe cells . The dervia these two receptors. Collectively, a better understanding of the differential roles of TNFRs may lead to the development of a treatment for glaucoma.Divergent actions of TNF in neurodegenerative diseases have been long implicated. Because of the opposite natures, we reasoned that both an inhibition of TNFR1 signaling and selective activation of TNFR2 signaling, respectively, could shift the balance of endogenous TNF activity toward an overall neuroprotective/regenerative response in retinal neurons . Since TCentral nervous system (CNS) is considered to be an immune-privileged system. It comprises a network of vessels with a very specialized endothelial lining supported by macroglia, microglial, pericytes, and neurons. Together these cells form a substantial blood-brain barrier (BBB). The latter strictly regulates the immune cells entering the CNS and maintains homeostasis concomitantly. However, under inflammatory conditions, the BBB gets disrupted, and immune cells migrate into the CNS parenchyma. Retinal neurons, including RGCs, are part of the CNS. They share several similar properties with other CNS neurons, including the inability to regenerate in response to cellular damage. The immune-mediated inflammation in the retina is tightly regulated by the specialized intraocular environment with tight interconnected junctions in a similar fashion to BBB, referred to as the blood-retinal barrier (BRB) . This baAccording to Streilein, the ocular immune-privileged is attributed to the immunosuppressive and anti-inflammatory properties of the ocular cells, tissues, and fluids . For insRecent studies have focused on immunological changes occurring in glaucoma pathogenesis, and potential preventative therapies based along those lines have been proposed. The major targets of interest are cytokines; as in the aforementioned, the inflammatory mediators play an essential role in the early progression of glaucoma and may regulate RGC survival or death , 40. In via the primary members of TNFR superfamily and high-affinity receptors; TNFR1 (p55) and TNFR2 (p75), that are expressed on different cell types and secreted predominantly by various inflammatory cells , Parkinson’s disease (PD), encephalopathy, meningitis, multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), myelin oligodendrocyte glycoprotein (MOG) antibody-associated disease, neuropathy as well as myelosuppression , 89, 90.Interestingly, patients with rheumatoid arthritis (RA) and psoriasis were reported with a low risk of developing AD when treated with TNF blockers therapy  , which sIt has been reported that there is an increased case of demyelinating disorders and opportunistic infections of CNS following TNF blockade. However, neurodegenerative diseases like dementia may also benefit from anti-TNF therapy. One of them is thalidomide, a drug that can reach CNS and block TNF and related cytokines local generation by microglia responsible for neuroinflammation . Due to in vivo models of glaucoma  has been investigated to deactivate astrocyte e injury . More stFurthermore, several TNFα signaling-associated biomarkers, such as NF-κB and MAPKs aim to govern signals of cell death or survival. More research is needed to investigate the neuroprotective effects of anti-TNF/TNFRs strategies in targeting axonal injury . The keyTNF inhibitors suppress the elevated TNF-mRNA found in the iris and ciliary body experimental autoimmune anterior uveitis . TopicalIn clinical trials of ocular inflammatory diseases, promising findings were reported on individuals with several phenotype uveitis effectively treated with adalimumab and infliximab –117. ThiInvestigating the anti-TNF agents has been growing as the subject of interest in the treatment of neurodegenerative diseases; however, some have shown no beneficial effects. For example, Infliximab, administered intraperitoneally 15 minutes after alkali burn inflicted, is demonstrated to provide significant retinal and corneal protection . Howeverin vivo glaucoma models. Although several studies documented contradicting role of TNFR1 and TNFR2 in glaucoma, TNF and its receptor may still serve as attractive targets in glaucoma therapy. For example, RGCs loss induced by TNF is shown to be caused by TNFR2, instead of TNFR1, when TNFR2 knockout mice showed no RGCs degeneration while in TNFR1 knockout mice showed otherwise .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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Nuclear morphological abnormalities, dysregulated gene expression, DNA repair deficiencies, telomere shortening, and genomic instability are all caused by progerin accumulation, all of which impair cellular proliferative capability. In addition, HGPS cells and preclinical animal models have revealed new information about the disease's molecular and cellular pathways and putative mechanisms involved in normal aging. This article has discussed the understanding of the molecular pathways by which progerin expression leads to HGPS and how the advanced therapy options for HGPS patients can help us understand and treat the condition.Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging condition that involves genetic mutations, resulting in debilitating phenotypic features. The present state of knowledge on the molecular pathways that contribute to the pathophysiology of HGPS and the techniques being tested  Hutchinson-Gilford progeria syndrome (HGPS) is a segmental "premature aging" condition in which children show phenotypes that may reveal information about the aging process at both the cellular and organismal levels . AccordiPathophysiological mechanism underlying the development of HGPSHGPS is a sporadic, autosomal dominant disorder caused almost entirely by de novo point mutations of the LMNA gene in codon 608 of exon 11 on chromosome 1 . The LMNProgerin presumably has a dominant-negative effect on the nuclear function of the LA-expressing cells. It is also hypothesized to harm other critical processes such as cell division, DNA replication, and gene transcription -15. CAAXClinical manifestations caused by HGPSTypically, the replicative capacity in an accelerated aging syndrome is reduced -26. AllsThe irregular telomere length regulation in a parental germ-line clone, an elevated cell turnover rate during progeric individual's growth, or an abnormally elevated rate of telomere deficit with every cell division could all contribute to progeric fibroblast's short telomeres . They coRigidity is a structural property that determines a bone's ability to withstand axial stresses, bending, and torsional moments . It is tProgerin accumulation can contribute to the development of thickening of the adventitia in small, medium, and large arteries, ultimately resulting in decreased intimal plaque formation potential, decreased vascular compliance, and increased vessel stiffness . As adveBy illustrating progerin exists in the coronary arteries of healthy aging individuals and rises with age, we also recognize a new factor in the traditional aging process -38. InhaManagement options for HGPSTo correct the defects in HGPS, a variety of treatment strategies have been proposed, including: (i) directly "repairing" the transmutation; (ii) inhibiting pre-mRNA mutant splicing generating progerin mRNA formation; (iii) reducing the toxic levels of isoprenylated and methylated progerin; (iv) to generate progerin removal; and (v) to reduce the harmful overdue buildup of progerin .Reduction of Progerin Downstream Toxic EffectsNuclear shape malformation, reactive oxygen species (ROS) production, aggregation of oxidized proteins, mitochondrial impairment, cell senescence, and NF-kB stimulation, resulting in elevating levels of secretion of the proinflammatory cytokines IL-6, CXCL1, and TNF-α, are just a few of the altered downstream pathways caused by progerin accumulation that have been described . In progTo investigate the curative prospect of NF-kB suppression on HGPS disease characteristics, Osorio FG et al. discovered that mating Zmpste24-/- mice alongside transgenic mice presenting decreased NF-B signaling improves lifespan and prohibits the progression of progeroid traits . They alAutophagy-activating DrugsRapamycin, an immunosuppressive drug used to prevent organ rejection, enhanced the aberrant nuclear shape, lagged the beginning of cellular aging in HGPS fibroblasts, and recovered the chromatin composition of fibroblasts in culture, involving BAF, LAP2alpha distribution patterns, and histone methylation status . OtherwiPrelamin A Isoprenylation and Methylation InhibitorsDuring posttranslational processing, the farnesylated carboxy terminus of prelamin A is removed due to the aberrant splicing event that gives rise to progerin . As a reDownregulation of Prelamin A Aberrant SplicingProgerin is secluded in atypically formed promyelocytic-nuclear bodies (PML-NB), which Harhouri K et al. discovered as novel progeria biomarkers . They diHumans with various LMNA transmutations that disrupt exon 11 splicings are referred to as "HGPS-like" patients . ProgeriThe introduction of CRISPR therapies has sparked hopes for a genetic approach by reducing progerin accumulation . A Cas9 HGPS is a segmental "premature aging" condition in which children show phenotypes that may reveal information about the aging process at both the cellular and organismal levels. Over the years, substantial progress has been made in understanding the disease, yet much more is to be learned. As the study progresses, HGPS continues to uncover previously unknown aspects of aging. With all of its complexities, HGPS provides a unique model for clarifying new lamin A/C and progerin roles in the cell. Progeria research has resulted in an increasing number of intriguing treatment possibilities. It is worth noting that in HGPS preclinical investigations, a drug's potential to selectively choose the cardiovascular system should be assessed, as it's the disorder's main functional aim, resulting in premature death. Cardiovascular measurements would also be selected among treatment efficacy readouts. In order to uncover new therapy pathophysiology targets and other outcome metrics, it will be necessary to improve our understanding of disease biology. This will allow us to understand better which damaged pathways are most relevant to the disease. We recommend that more future studies be performed to uncover the treatment efficacies of current and upcoming management modalities.


Cancer recurrence and metastasis, following successful treatment, constitutes a critical threat in clinical oncology and are the leading causes of death amongst cancer patients. This phenomenon is largely attributed to metastatic tumor dormancy, a rate-limiting stage during cancer progression, in which disseminated cancer cells remain in a viable, yet not proliferating state for a prolonged period. Dormant cancer cells are characterized by their entry into cell cycle arrest and survival in a quiescence state to adapt to their new microenvironment through the acquisition of mutations and epigenetic modifications, rendering them resistant to anti-cancer treatment and immune surveillance. Under favorable conditions, disseminated dormant tumor cells ‘re-awake’, resume their proliferation and thus colonize distant sites. Due to their rarity, detection of dormant cells using current diagnostic tools is challenging and, thus, therapeutic targets are hard to be identified. Therefore, unraveling the underlying mechanisms required for keeping disseminating tumor cells dormant, along with signals that stimulate their “re-awakening” are crucial for the discovery of novel pharmacological treatments. In this review, we shed light into the main mechanisms that control dormancy induction and escape as well as emerging therapeutic strategies for the eradication of metastatic dormant cells, including dormancy maintenance, direct targeting of dormant cells and re-awakening dormant cells. Studies on the ability of the metastatic cancer cells to cease proliferation and survive in a quiescent state before re-initiating proliferation and colonization years after successful treatment, will pave the way toward developing innovative therapeutic strategies against dormancy-mediated metastatic outgrowth. Cancer progression largely comprises the processes of primary tumor growth and secondary metastasis. More specifically, it refers to the phenotypic changes in already-formed neoplastic lesions, including morphological, molecular, and functional changes in tumor cells while incorporating an extensive variability of signals and mechanisms  ImportanRegarding the metastatic potential of cancer cells, there are various molecular and cellular mechanisms underlying the different steps of the metastatic cascade, including the epithelial-to-mesenchymal transition (EMT), invasion, anoikis, transport through vessels, the outgrowth of secondary tumors, and angiogenesis through neovascularization . NeovascThere are some intriguing similarities between the concept of cancer stem cell (CSC) theory and cancer cell dormancy, including their contribution to disease relapse due to their drug-resistant properties. However, they have some key differences. Dormant cancer cells are known to enter a quiescent state characterized by a reversible, cell cycle growth arrest, while CSCs growth is only delayed in the cell cycle . AsymmetAdditionally, DTCs enter metastatic dormancy in order to adapt to either the bloodstream, the TME, or the target organ  18]. Th. Th18]. In respond to microenvironmental cues, DTCs can re-enter the cell cycle and adjust to their new microenvironment. Thus, it is of paramount importance to understand the biology of DTCs reawakening in order to prevent metastatic outgrowth. In this article, we aim to comprehensively describe dormancy and escape of dormancy mechanisms and explore how the dormant disseminated cells, which are widely understood to be the seeds of metastasis, could represent an effective approach towards limiting the expansion of secondary lesions, as well as how targeting this stage of metastatic spreading may provide a potential therapeutic approach.Dormancy and its maintenance are complex processes, mediated by intrinsic and autocrine signaling, and it is also controlled by signals derived from immune cells, endothelial cells, fibroblasts and other non-cellular components of the tumor microenvironment ,24. In tDTCs moving from the primary site to a distant site remain in a dormant state by escaping the proliferative cycle or by reaching an equilibrium between proliferation and apoptosis. While being in a dormant stage, dormant DTCs can accumulate additional molecular, epigenetic and genetic aberrations that enable them to optimally adjust in their new microenvironment. The intrinsic mechanisms described in this review refer to the genetic and epigenetic, apoptotic, proliferative, survival and autophagy-related alterations and intracellular signaling .A correlation has been demonstrated between various alterations in the genome, primarily linked to cell differentiation/proliferation and the induction and maintenance of the dormant phenotype as shown in several preclinical cancer models. Overexpression of the FBXW7 gene has been rendered a crucial component for the maintenance of breast cancer dormancy. In the presence of FBXW7, entrance to the cell cycle is restrained through the ubiquitination and subsequent proteasomal degradation of its inducers c-Myc and cyclin E. When genetically ablated, the FBXW7 gene led to not only the disruption of the quiescent state of DTCs, but also the initiation of their proliferation in both allograft and xenograft models. DTCs lacking FBXW7 were significantly depleted following paclitaxel treatment, thus indicating increased sensitivity to chemotherapy. Poorer prognosis and shorter cell survival was associated with tumors expressing high levels of FBXW7 compared to FBXW7-deficient cells following tumor resection and chemotherapy treatment .A dormant phenotype of breast DTCs in the bones has been linked to the leukemia inhibitory factor receptor (LIRF) which interacts with the interleukin-6 (IL-6) cytokine family. LIRF acts as a suppressor of cytokine signaling (SOCs) and an activator of signal transducer and activator transcription 3 (STAT3). Upon LIFR ablation, downregulation of dormancy-associated genes, reactivation of proliferation and bone colonization was observed . AnotherAutophagy is another proposed crucial mechanism for the re-activation, survival and adaptation of dormant DTCs. Autophagy is an evolutionary conserved, physiological mechanism of fitness and cell survival, whose activation highly depends on the presence of unfavorable, metabolic stress conditions, such as nutrient deprivation. Its mechanism of action is associated with the degradation and subsequent recycling of damaged cytosol components, misfolded proteins, organelles and other macromolecules in order to sustain homeostasis and diminish cell damage. As such, evidence begins to accumulate regarding the exploitation of autophagy by tumor cells to endorse their survival under oxidative stress, improve their bioenergetics and thus facilitate tumor progression  30]. Pr. Pr30]. 3K/AKT/mTOR) . In. In34]. henotype .Cancer dormancy can be separated into cellular dormancy and tumor mass dormancy (rate of proliferation rate = apoptosis rate). Many intracellular factors can regulate tumor dormancy. An example of glioblastoma dormancy regulators include the epidermal growth factor receptor (EGFR) and thrombospondin (TSP) . Other t3K)/AKT signaling pathway, comprising the extracellular stress protein clusterin and IGF1, under external stimuli is another mechanism thought to be involved in the switching between cancer cell proliferation and dormancy  under serum deprived conditions  breast cancer cells is NFkB. Following the expression of a ‘constitutively active form of IkB kinase b (CA-IKKβ)’ in cell lines, a reversible inhibition of E2-dependent cell proliferation and tumor growth was demonstrated in vitro and in vivo. Similarly, co-activation of IKKβ and ER enhanced cell invasion and migration to guide the experimental development of metastasis . Decreasdormancy . The negnditions . Cell prFluctuations between a proliferating and dormant state of DTCs could also be the consequence of epigenetic reprogramming mechanisms, encompassing transcriptional regulation through changes in chromatin structure . Some of+ breast cancer patients, the MSK1 kinase has been identified as another crucial regulator of metastatic dormancy, since its higher expression levels correspond to lower probability of developing early metastases. Breast cancer cell differentiation is impaired through the downregulation of MSK1, therefore enhancing their growth potential and bone metastases. The expression of the transcription factors FOXA1 and GATA3, which are vital for cell differentiation, is also regulated by MSK1 depending on the alterations of their chromatin status . Al. Al24]. Adaptive ImmunityT cell content and immune cell activation in tumors in the bone marrow of cancer patients was significantly increased compared to healthy individuals, indicating signs of tumor dormancy. In other words, the bone marrow represented a prosperous site for the presence of lethal DTCs in a dormant state with a potential contribution to this by the high presence of T cells .+ T-cells, the depletion of which reversed cancer cell dormancy . Th. Th+ T-cCellular dormancy could also be influenced by immune evasion. In a model of acute myeloid leukemia, overexpression of the programmed death-ligand 1 (PD-L1)  led to cytotoxic T cell-mediated resistance to killing and thus a potential contribution dormancy . Dormanc+ T cell mediated cytotoxicity. Escape from dormancy and an increase in proliferation was noted after depletion of CD4+ and CD8+ T cells in a pancreatic cancer mouse model (RIP-Tag2) . Mo. Mo50]. Another therapeutic approach focuses on increasing susceptibility of dormant cells to anti-cancer treatment by disturbing their dormant state and ‘re-awakening’ them . Dormant (DTPs) numbers in various cancer cell lines that have undergone chemotherapy treatment. Successful DTC killing by CPI-455 was shown in breast, colon, melanoma and lung cancer models [Another therapeutic approach aims at directly targeting dormant cells to reduce the possibility of future disease relapse  10]. A . A 10]. r models . Furtherr models . A recenr models . Death or models . In addiTherapeutic targeting of dormant DTCs has therefore attracted significant interest and constitutes a promising avenue for improved clinical outcomes of cancer patients. Unfortunately, due to the immense amount of information missing regarding cellular or surface markers that could lead to the identification of DTCs as well as limitations in current diagnostic technologies, this therapeutic strategy is still very challenging . A permaCellular dormancy can be studied in vitro owing to the development of a platform using a method known as ‘cancer-tissue originated spheroid method (CTOS)’, which allows for hypoxic conditions to be applied for a minimum of 7 days without requiring stimulation by growth factors . Novel t3K/AKT and p38 expression . Th. Th68]. There is a perplexing inconsistency between the lack of completely malignant DCCs during a surgery performed in order to remove a primary tumor, their less frequent detection compared to the extremely rare ‘immature’ DCCs, and the apparent genomic parallel between some primary tumors and obvious metastasis. Thus, there is a question mark whether metastasis originates by early DCCs or late DCCs. Early DCCs need to evolve and obtain significant changes at the distant site, whereas late DCCs can find metastasis after adapting to the new environment followed by clonal development. Studies in animal models suggest that a percentage of 80% of metastasis cases derived from early DCCs, while on the other hand genomic analysis of DCC together with the primary tumor and metastasis has showed contradicting conclusions. When DCCs are isolated from patients without obvious metastasis, their genomic characteristics reveal an early genomic departure from the key clone of the primary tumor, indicating dissemination early in ‘genomic’ time ,165,166.In this article, we reviewed the signaling pathways, the intrinsic and extrinsic mechanisms that either sustain dormancy, making DTCs more challenging to identify and eradicate, or enable the escape of these cells from their dormant state at distal sites, causing metastasis. Most of the known signals and mechanisms are characterized as organ-specific, suggesting that dormancy is regulated through the tissue microenvironment and that pharmacological targeting of cancer cell dormancy might involve a highly personalized approach. Thus, targeting DTCs has been increasingly gaining attention. DTCs self-seed for prolonged periods of time at their dormant state, giving scientists the opportunity for therapeutic intervention and experimentation. A promising approach for the complete eradication of DTCs is by targeting both proliferating and dormant cells by using a combination of drugs that will also prevent cancer recurrence. Remarkably, several studies support the sequential combination of drugs rather than co-treatment. There is an obvious need for improving preclinical dormancy models and developing sensitive diagnostic imaging tools that can detect dormant cancer cells. Finally, designing new dormancy-targeting therapeutic drugs to overcome resistance to metastatic cancer therapies is urgently needed.


Cannabis sativa has caught the attention of health-conscious consumers and the food industry. Historically, seeds have long been utilized as a food source and currently there is an increasing number of edibles on the market that contain cannabis. Cannabinoids include the psychoactive constituent, delta-9-tetrahydrocannabinol (THC), and the non-psychoactive cannabidiol (CBD) that are both compounds of interest in Cannabis sativa. This paper looks at the distribution of nutrients and phytocannabinoids in low-THC Cannabis sativa, the historical uses of hemp, cannabis edibles, and the possible side-effects and concerns related to cannabis edibles. Several authors have pointed out that even though the use of cannabis edibles is considered safe, it is important to mention their possible side-effects and any concerns related to its consumption that negatively influence consumer acceptance of cannabis edibles. Such risks include unintentional overdose by adults and accidental ingestion by children and adolescents resulting in serious adverse effects. Therefore, cannabis edibles should be specifically packaged and labelled to differentiate them from known similar non-cannabis edibles so that, together with tamperproof packaging, these measures reduce the appeal of these products to children.The published health benefits of  Food products infused with chemical compounds that possess health benefits beyond basic nutrition  are piquCannabaceae family [Cannabis sativa, Cannabis indica [Cannabis ruderalis recognized as the three main species in cannabis [Cannabis sativa contains an abundance of non-nutritive, bioactive phytochemicals known as phytocannabinoids, with delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) as phytocannabinoids of particular interest. Cannabis sativa plants containing less than a specified amount of THC in dried weight are also known as hemp. In certain European countries, THC levels should not exceed 0.2%. In the United States of America (USA) [Cannabis sativa plants containing more than the specified THC limit (high THC or marijuana) are considered as schedule 1 drugs [Cannabis is a generic term describing an annual herbaceous plant that belongs to the e family ,6 with Cs indica  and, thocannabis . Cannabica (USA) ,11 and Cca (USA) , THC levca (USA) . Cannabi 1 drugs ,15. Ther 1 drugs .Cannabis sativa (or hemp plants) and associated cannabinoid CBD as a functional ingredient in food products.Those who use the aerial parts of the marijuana plants, such as the leaves and flowers, became intoxicated after smoking and when smoking on a regular basis, may have a notable increase in the incidence of health risks such as mental illnesses . As a reCannabis sativa, or hemp, plants, despite a considerable amount of scientific proof [Many consumers are uninformed regarding the nutritional benefits associated with ic proof ,23,24. Fic proof ,26. The ic proof ,28,29.Proteins found in the hemp seed are legumin-type globulin edestin (67 to 75%) and globular-type albumin (25 to 37%)  but, likAround 110 different cannabinoids have been identified in cannabis ,33. CannCannabis sativa subsp. indica or marijuana) where humans smoke a dry combination of its shredded leaves, stems and flower buds [Archaeological findings indicate that cannabis has been cultivated for over 6000 years, possibly originating in China . The moswer buds . This hawer buds  consequewer buds . Figure wer buds ,11,20,44wer buds ,44,45,46wer buds ,47.The hemp seed is achene : containThe edible forms of hemp seeds include hulled seed, hempseed oil and its crushed by-products, hemp seed cake, or meal . Hemp seMore recently, hemp seeds have proven to be a versatile ingredient in food production  with theCannabis arrived in Africa about 1000 years ago and has become an important source of income for rural communities . HoweverThe concept of nutrition has reached a greater audience because of reports describing diet- and lifestyle-related non-communicable diseases . As a re®) and claiming CBD as a dietary supplement is currently illegal in the United States [Apart from the seeds, the inclusion of cannabinoids such as CBD in functional foods could support health and reduce the risk of disease. Food products that contain cannabinoids such as CBD are referred to as cannabis edibles and include a range of products from baked goods and sweets to beverages ,67. Thoud States .Edibles usage is classified into medicinal and recreational purposes . About 1South Africa permits some general, low-risk health maintenance or health enhancement claims on sold CBD products. However, the permitted amount of CBD per serving should not exceed 20 mg and the sales pack may not exceed a total of 600 mg of CBD  and shouJust as smoking cannabis may have side-effects, consuming edibles may also have pharmacokinetic effects  whose syCannabidiol (CBD) and CBD-products for private use are increasing in popularity amongst European , South ASecondly, there is concern regarding either intentional or accidental adulteration or contamination of cannabis edibles that might be harmful to the consumer, including heavy metal and pesticide contamination , contamiA major concern surrounds pediatric exposure to cannabis edibles . A studyWith this range of concerns regarding the ingestion of cannabis edibles from production lines that may have lax quality assurance measures leading to the production and sale of cannabis edibles with unwanted constituents, it is essential that legislation prescribing cannabis product preparation and labelling should be clear and enforceable .Cannabis sativa possesses many health-promoting qualities and so it has played an effective role as a traditional medicine to treat a variety of ailments from pain, anxiety and weight gain through to conditions such as cardiovascular disease and diabetes, as well as infectious diseases such as malaria, and cancer. Opinion regarding cannabis edibles is changing amongst consumers and most countries around the world are shifting towards the legalization of the recreational and medicinal use of cannabis leading to a rapid increase in the global acceptance and availability of cannabis edibles. A wide range of products infused with cannabis extracts are currently available on the market and, unfortunately, this increases the possibility of side-effects to the consumer. Not only are most cannabis edibles not regulated, but there is risk of accidental ingestion of cannabis, particularly by children. Such incidences are on the rise in those countries where cannabis has been legalized or decriminalized. Most cannabis edibles have not been approved by the FDA but are currently subject to evaluation regarding long-term safety and cumulative effects in humans. This calls for efficient and effective safety measures to prevent pediatric cannabis poisoning and accidental unintended high when amending policies for cannabis usage. In addition, there must be some form of commercial and consumer awareness regarding the optimal preparation and packaging of cannabis edibles to promote their safe and enjoyable consumption.