Treatment of locally advanced rectal cancer has been subject to pronounced changes based on the results of recent prospective trials. New paradigms have been introduced, including the shift of systemic treatment components from adjuvant to neoadjuvant setting , the omission of surgery in patients with clinical complete responses after neoadjuvant treatment (non-operative management) and the introduction of upfront immunotherapy in patients with microsatellite instability (MSI)-high/mismatch-repair-deficient (dMMR) tumors. We developed an institutional treatment algorithm which may serve as a practical tool for treating physicians without any claim to general validity.Total neoadjuvant therapy (TNT)—the neoadjuvant employment of radiotherapy (RT) or chemoradiation (CRT) as well as chemotherapy (CHT) before surgery—may lead to increased pathological complete response (pCR) rates as well as a reduction in the risk of distant metastases in locally advanced rectal cancer. Furthermore, increased response rates may allow organ-sparing strategies in a growing number of patients with low rectal cancer and upfront immunotherapy has shown very promising early results in patients with microsatellite instability (MSI)-high/mismatch-repair-deficient (dMMR) tumors. Despite the lack of a generally accepted treatment standard, we strongly believe that existing data is sufficient to adopt the concept of TNT and immunotherapy in clinical practice. The treatment algorithm presented in the following is based on our interpretation of the current data and should serve as a practical guide for treating physicians—without any claim to general validity. For two decades, locally advanced rectal adenocarcinoma was approached by neoadjuvant short- or long-course radiation therapy (RT) with or without concomitant CHT followed by surgery and—in case of pathologic nodal involvement—adjuvant CHT. This approach was based on proven efficacy and acceptable toxicity in large, randomized trials ,2,3. TheRecently, two large randomized phase III trials have reported significantly improved distant failure and disease-free survival (DFS) rates using either induction or consolidation CHT sequential to (chemo)radiation compared to the standard approach ,10. AsidHowever, aside from similar major findings, the mentioned trials still considerably differed regarding the used (chemo)radiation regimen and several inclusion criteria. Especially the risk profile for locoregional and/or distant failure based on certain prognostic factors at presentation, such as mesorectal fascia involvement (MRF+), extent of nodal involvement , or the presence of extramural vascular invasion (EMVI), was not uniform across the trials. This results in some uncertainty regarding the optimal treatment approach for different risk groups of patients: who should be scheduled for a TNT approach per se, which (chemo)radiation regimen should be included for whom, who should be selected for organ preservation, and how should the follow-up be performed in these patients. At our institution, changes in the general treatment algorithm were preceded by intensive, structured multidisciplinary discussions of the available data including specialized surgeons, radiation and medical oncologists, radiologists, and pathologists, resulting in a comprehensive consensual new treatment algorithm. The current manuscript aims at presenting this algorithm including its rationale based on our interpretation of the available literature.Within a TNT and/or NOM concept, initial staging gains even more importance for patient selection due to the transfer of all additional treatments into the preoperative phase and the increasing number of prognostic factors with relevance for treatment decisions. The most relevant factors for defining risk group and treatment strategy include T and N stage, depth of infiltration into the mesorectal tissue, involvement of the mesorectal fascia, presence of lateral  suspicious lymph nodes, presence of EMVI, and tumor distance from the anal verge. All the mentioned factors are known to be associated with the risk of either locoregional recurrence or distant metastases or (mainly) both ,19,20,21Based on current recommendations, these factors are determined most accurately by a combination of pelvic MRI and rigid endoscopy with endorectal ultrasound (EUS), flanked by CT of chest and abdomen or FDG-PET-CT to rule out distant metastases . While ERegarding response evaluation, standard MRI alone lacks accuracy for precisely defining ycT and ycN stage  and pred®, Best, The Netherlands). We routinely use spasmolytics such as butylscopolamin or glucagon to reduce artefacts from peristalsis, but discourage the routine use of rectal fillings due to possible misinterpretation of the distance between the tumor and the mesorectal fascia with the risk of overstaging [Technically, there is broad consensus that T2 weighted sequences acquired in three planes including thin axial sections (slice thickness 3 mm) perpendicular to the long axis of the rectal wall at tumor location build up the basis for accurate rectal cancer staging ,28. Althrstaging ,30.Regarding the detection of distant metastases, current guidelines recommend the use of chest and abdominal CT ,24. FDG-In summary, we strongly acknowledge the importance of extensive primary staging and sophisticated response evaluation in the decision-making process. We therefore decided to perform all staging procedures  and the final response assessment after completion of neoadjuvant treatment preferably at our tertiary cancer center. If a NOM approach is used, all follow-up examinations for the first two years are also scheduled at our institution. We further agreed on criteria for the standardized assessment and reporting of pelvic MRI during staging, response assessment, and follow-up together with our radiologists. These criteria are mainly based on the ESGAR recommendations  for MRI Our TNT-approach is mainly based on the regimens used in the RAPIDO- (short-course radiation therapy (RT) followed by consolidation CHT)  and OPRAPatients with high-risk factors;Patients with distal tumors;Patients with intermediate-risk factors and non-distal tumors.In our opinion, multimodal treatment should follow two major rules: risk-adaption and preservation of quality of life, leaving us with three major patient groups: We defined high-risk features on pelvic MRI according to the inclusion criteria of the RAPIDO trial : cT4 staBecause these patients are at the highest risk for locoregional and/or distant failure, we offer them the most intensive therapy within our TNT concept, namely long-course CRT followed by consolidation CHT  and surgPatients with distal tumors are at the highest risk for the need of an abdominoperineal resection with permanent colostomy, limiting their quality of life. Moreover, they face a considerable risk of functional impairment even after sphincter-saving procedures known as low anterior resection syndrome (LARS). A recent meta-analysis evaluated 36 trials and quantified the risk for LARS after low anterior resections as high as 44% . Thus, pWe therefore offer all patients with distal tumors the most intensive therapy within our TNT concept, namely long-course CRT followed by consolidation CHT and NOM if a cCR is achieved . In patiExcluding patients with high-risk features and distal tumors leads to a third group of patients suffering from rectal cancer with “intermediate”-risk features located in the middle and upper third of the rectum. Several non high-risk but still prognostic factors exist to guide treatment decisions in those patients. Tumor location in the upper third as a factor has been a matter of debate since the introduction of neoadjuvant treatment, based on the different definitions of rectal length in US and European trials . Data onAnother factor predicting the outcome is the depth of mesorectal invasion. This knowledge has led to a subclassification of T3 stage  into four stages: T3a (<1 mm invasion), T3b (≥1–5 mm), T3c (>5–15 mm), and T3d (>15 mm depth of invasion) . While tLymph node status represents one of the oldest but most debated risk factors in rectal cancer. Its general impact on outcome seems crystal-clear based on large, pooled analyses ,46 and pWhile overtreatment may represent a considerable risk, undertreatment caused by inaccurately described nodal negativity might be even worse. Several analyses have shown rates of pathologically confirmed positive nodes in 20–30% of patients initially staged as cT3cN0 on EUS and MRI with or without neoadjuvant treatment ,48,49. IWhich specific parameters to define a local cCR on imaging and endoscopy should be used?Which is the best timing for response assessment?Do we attempt a histological “confirmation”  of a cCR?Similarly to the increasing importance of pre-treatment evaluation for the indication of neoadjuvant therapies since the introduction of TNT concepts, response evaluation has gained critical importance in the view of NOM. As NOM-approaches based on current data only seem to be safe after achieving a cCR , all posThe general advantages and disadvantages of different MRI sequences in response assessment have already been discussed above. In the absence of clear evidence  favoring certain parameters for classification of a response as “complete” or the prediction of a pCR based on MRI and/or endoscopy, we decided to simply follow the criteria for cCR of the Memorial Sloan Kettering Cancer Center (MSKCC) regression scheme used in the OPRA trial  ,31 as thThe optimal time interval for response assessment is highly debated. For non-TNT approaches, there is increasing evidence that longer time intervals after (chemo)radiation (>8 weeks) may increase the likelihood of pCR without facing higher postsurgical morbidity ,51. In cTo ensure maximum safety for patients within NOM approaches, some centers advocated (blind or guided) biopsies of the primary tumor region to “confirm” a complete remission in contrast to relying only on endoscopy and imaging. After multidisciplinary discussion, we decided not to perform such biopsies for the following reasons: We offer a NOM approach only in case of a cCR of primary tumor and suspicious nodes (ycT0ycN0). As the latter are mostly not amendable to biopsy, we will have to rely on imaging in most patients anyway. Moreover, the combination with endoscopy already increases safety with regard to possible superficial residuals of the primary tumor, and blind biopsies of the tumor region in case of complete remissions seem generally flawed by the risk of geographic misses . BecauseOur follow-up procedure generally depends on whether planned surgery has been done or if the patient is part of a NOM approach. Surgically treated patients are followed according to institutional standards based on current guidelines, which will not be elaborated any further. For patients within a watch and wait policy after cCR, we opted for a very close follow-up scheme . This scAround 5 to 10% of rectal cancers show microsatellite instability (MSI-high) or mismatch-repair–deficiency (dMMR) . This suRecently, first data have been presented for treating locally advanced rectal cancer patients with the anti-PD-1 antibody dostarlimab : in a siAlthough these results have to be confirmed in larger trials before entering clinical routine, we believe that in the light of these data, treatment with immune checkpoint blockade should be discussed with patients with MSI-high/dMMR rectal cancer on an individual basis, especially when aiming for an organ-sparing strategy. We strongly encourage referring these patients to centers with respective clinical trials, in order to contribute to the body of knowledge that will eventually lead to the approval of these drugs and thereby the possibility of universal employment in all patients with MSI-high/dMMR disease.In light of increased pCR rates and improved DFS, we have implemented total neoadjuvant therapy (TNT) as institutional standard for treating patients with locally advanced rectal cancer in the presence of high-risk features. Furthermore, we routinely offer TNT to patients with distal rectal cancer with the goal of non-operative management. Whenever TNT is indicated, immunotherapy should be considered in case of microsatellite instability. The presented treatment algorithm is based on our interpretation of the current data and should serve as a practical guide for treating physicians—without any claim to general validity.


The stretching of a myopic eye is associated with several structural and functional changes in the retina and posterior segment of the eye. Recent research highlights the role of retinal signaling in ocular growth. Evidence from studies conducted on animal models and humans suggests that visual mechanisms regulating refractive development are primarily localized at the retina and that the visual signals from the retinal periphery are also critical for visually guided eye growth. Therefore, it is important to study the structural and functional changes in the retina in relation to refractive errors. This review will specifically focus on electroretinogram (ERG) changes in myopia and their implications in understanding the nature of retinal functioning in myopic eyes. Based on the available literature, we will discuss the fundamentals of retinal neurophysiology in the regulation of vision-dependent ocular growth, findings from various studies that investigated global and localized retinal functions in myopia using various types of ERGs. The prevalence of myopia is on the rise worldwide since a few decades . VariousThe development of neural retina usually begins on day 26 of gestation , where tRetina being the only photo-sensitive neural layer in the eye , incorpoSeveral animal species including chicks –44, squi−/− mice with rod dysfunction . T. T95]. TThe results from several studies that investigated mfERG responses in myopia are given in Table Previous studies have found that the mfERG amplitude density are maximum, and the latencies of P1 and N2 waves are the longest at the central R1 (fovea), which progressively decreased with the increase in retinal eccentricity , 148. HoThere are various other factors such as optical, electrical, and retinal factors that contribute to the reduced mfERG responses. For optical factors, the degraded mfERG responses are associated with the decreased retinal image size and retinal illuminance due to elongated axial length in myopes. Electrical factors such as increased electrical path and ocular resistance from the electrical sources  and the ERG electrodes  and lowe2 and 7.4 nV/deg2 in N1 and P1 amplitude density, respectively, in an absence of a myopic retinal degeneration [Similar to ffERG, the mfERG amplitude density also shows a better correlation with axial length compared to refractive error , 110, whneration . High myneration , and outneration , 158. Thneration .Given that the mfERG responses primarily arise from retinal bipolar cells and nuclear layer, any physical or mechanical alterations to these cells will have an impact on mfERG responses . Hood etThe mfERG responses gradually decline toward the periphery  and corrPrevious studies have shown an increase in the central subfield macular thickness (CST)  and a reThe PERG is a contrast-based response, driven by macular photoreceptors and originating from retinal ganglion cells . It is aThe P50 and N95 wave amplitudes of the transient PERG response were reduced in high myopes with longer axial length compared to that of emmetropes and low myopes , 165–167A further refinement of mfERG is the gmfERG, in which a successive insertion of a full-field or global-flash stimulus between consecutive focal flashes of a standard mfERG stimulus enhances the adaptive response, isolating the outer and inner retinal responses into two major components , 169. ThThe results from several studies that investigated gmfERG in myopia are given in Table The gmfERG on myopic children with different contrast levels exhibited a significant reduction in central macular (R1) DC amplitude density at 96% contrast, while the IC amplitude density was unaffected . But myoA similar contrast-based gmfERG setup was used to determine whether myopia progression measured over 1 year was associated with changes in retinal function. At 49% contrast, both the DC and IC amplitude densities at the macula  were significantly reduced with the progression of myopia –178. TheGiven an alarming rise in the prevalence of myopia worldwide, various optical, pharmacological, and environmental strategies are being incorporated to prevent the development of myopia  as well Besides optical anti-myopia strategies, pharmacological management of myopia progression with atropine eye drops has also been one of the most effective strategies to control myopia progression in children , 189–193To summarize, there are significant changes in retinal function, as assessed from ERG testing in myopes, and these changes strongly correlate with axial length and the degree of myopia. Although some investigations with the ffERG show significantly reduced dark-adapted and light-adapted a- and b-wave amplitudes with increasing degree of myopic refractive error, there is some evidence that dark-adapted responses are further attenuated than the light-adapted responses. These findings suggest that myopia may be associated with reduced photoreceptor (mainly rod response) and ON-bipolar cell activity. Several studies with the mfERG show reduced P1 amplitude density in myopes, suggesting alterations in retinal ON-and-OFF cone bipolar cells in myopia. The mfERG amplitude density associated with refractive error varies significantly with retinal eccentricity , axial length, and the degree of myopia. Finally, studies have reported reduced PERG amplitudes and gmfERG amplitude density in both the central and paracentral retina in high myopia. While these studies illustrate important associations between myopic refractive error and changes in retinal electrical activity, there has been limited work to understand the longitudinal changes in the ERG and how they relate to myopia progression in younger eyes. Future work investigating electrophysiological responses in combination with the measurements of retinal structural changes  will provide valuable insights into how retinal electrical changes may influence ocular growth and refractive error development in humans. Given the availability and wide use of optical and pharmacological anti-myopia management strategies that are known to act at retinal level , it would also be interesting for future studies to examine how these anti-myopia interventions interact with retinal signals to prevent myopia.


Acute respiratory distress syndrome (ARDS) is a major concern in critical care medicine with a high mortality of over 30%. Injury to the lungs is caused not only by underlying pathological conditions such as pneumonia, sepsis, or trauma, but also by ventilator-induced lung injury (VILI) resulting from high positive pressure levels and a high inspiratory oxygen fraction. Apart from mechanical factors that stress the lungs with a specific physical power and cause volutrauma and barotrauma, it is increasingly recognized that lung injury is further aggravated by biological mediators. The COVID-19 pandemic has led to increased interest in the role of the renin-angiotensin system (RAS) in the context of ARDS, as the RAS enzyme angiotensin-converting enzyme 2 serves as the primary cell entry receptor for severe acute respiratory syndrome (SARS) coronavirus (CoV)-2. Even before this pandemic, studies have documented the involvement of the RAS in VILI and its dysregulation in clinical ARDS. In recent years, analytical tools for RAS investigation have made major advances based on the optimized precision and detail of mass spectrometry. Given that many clinical trials with pharmacological interventions in ARDS were negative, RAS-modifying drugs may represent an interesting starting point for novel therapeutic approaches. Results from animal models have highlighted the potential of RAS-modifying drugs to prevent VILI or treat ARDS. While these drugs have beneficial pulmonary effects, the best targets and application forms for intervention still have to be determined to avoid negative effects on the circulation in clinical settings. The renin-angiotensin system (RAS) plays a role in many cardiovascular, renal, and pulmonary processes. It is a network of peptides  that areMechanical ventilation is required in clinical conditions associated with acute pulmonary gas exchange deterioration, reduced vigilance, and loss of protective reflexes. The unphysiological state of positive pressure ventilation subsequently impacts the body further. The relevance of this impact increases with higher settings of mechanical power variables such as ventilation pressure, tidal volume and respiratory rate . ComponeThe RAS and potential sites of intervention have been studied in various experimental settings, sometimes yielding conflicting results that depend on the exact parameters of the experimental models. Ventilation with a very high tidal volume of 40 ml/kg in rats led to increased protein content in BALF, pulmonary expression and serum levels of MIP-2, and lung tissue levels of Ang II after 4 h, while mRNA expression levels of ACE2 were decreased . The effThe role of ACE activity in VILI is somewhat controversial and may depend on the exact parameters of the model system, the biological compartments that were investigated, and the analytic methods. In mechanically ventilated rats with tidal volumes of 18 ml/kg aiming at moderate alveolar hyperdistension, lung ACE activity was significantly decreased . In a moAge is an important factor for interpreting the results of animal lung injury models. In a systematic review, 2 = 12%) as well as primary murine alveolar epithelial type II cells and human small airway epithelial cells exposed to 1% oxygen have demonstrated a hypoxia-induced increase in ACE2 mRNA expression  and product (Ang II) is prevented with protease inhibitors. Ang II is quantified by LC-MS/MS and the specific activity of ACE is calculated by determining the inhibitor-sensitive fraction of Ang II formation. The active ACE concentration is then calculated by relating the ACE activity in the sample to the activity of recombinant human ACE in plasma. The assay to determine active ACE2 follows an equivalent process, namely spiking Ang II, application of the ACE2-specific inhibitor MLN-4760, and quantification of Ang 1–7. The Ang 1–7 formation rate is calibrated to a standard curve of ACE2 from healthy human control plasma. With the help of this method, it was discovered that high concentrations of active ACE2 in the plasma of hospitalized patients with COVID-19 depend on disease severity . IncreasIn light of the COVID-19 pandemic, interest in the function of the RAS in the context of ARDS has grown, as ACE2 serves as the primary cell entry receptor for SARS-CoV-2 . IncreasAt first glance, results from studies measuring circulating angiotensin peptides and soluble ACE2 in patients with COVID-19 do not completely fit this theoretical model of increased Ang II and downregulated ACE2 levels. A study of Ang II serum levels in a cohort of 112 patients with COVID-19 reported a decrease in Ang II levels that was more pronounced in patients with ARDS and in non-survivors . AnotherAngiotensin converting enzyme polymorphisms contribute to the relatively high standard deviation of Ang II plasma levels in patient cohorts and are associated with the severity of ARDS . From th2/FiO2 ratio were higher in patients treated with enalapril than in the control group.An early randomized controlled trial on the hemodynamic and respiratory effects of enalapril in 45 critically ill patients was published in 1995 . While eclinicaltrials.gov as NCT04335136), but results are yet to be published. A case report on rhACE2 treatment in a patients with severe COVID-19 confirmed the intended effect of a reduction in Ang II levels during twice daily intravenous treatment with rhACE2 for 7 days (After a successful phase I study of pharmacokinetics and pharmacodynamics of recombinant human (rh)ACE2 in healthy volunteers , a clinir 7 days .There is evidence that ACE inhibitors protect against radiation pneumonitis in lung cancer patients . This inExperimental and clinical evidence shows that the RAS is involved in VILI and other types of pulmonary inflammation where it may offer novel therapeutic targets. Despite the increasing precision and detail of describing RAS activation with mass spectrometry-based assays, many aspects of RAS enzyme activity remain elusive. ACE inhibitors, angiotensin receptor blockers and Ang 1–7 supplementation seem to protect from VILI in experimental studies in rodents, not only by affecting levels of RAS components but also by acting on proinflammatory cytokines such as IL-6 and MIP-2, and there is evidence of decreased histological lung injury . Howeverclinicaltrials.gov aim to include patients with COVID-19 and to test a treatment with angiotensin receptor blockers, ACE inhibitors, or Ang 1–7. Apart from selecting the most suitable substances, the challenge is to establish the most effective form of application. Since the lungs can be reached by circulation, systemic application of RAS-modifying drugs has a substantial impact on angiotensin metabolite concentrations in plasma (Another broad field for future study is applying the findings about involvement of the RAS in ARDS in randomized controlled clinical trials. Various trials currently listed in n plasma . Inhalatn plasma .In summary, knowledge about RAS activation in experimental lung injury and clinical ARDS is increasing in quantity and detail. The mechanisms of regulating RAS enzyme activities and their shedding remain elusive, especially in the clinical setting, and require further study with innovative measurement tools. However, even with the current conception of RAS activation in ARDS, clinical studies can be designed to counteract primary and ventilator-induced lung injury. Several clinical trials with RAS-modifying drugs are currently underway for COVID-19, with most results still pending.KK drafted the manuscript and prepared the figures. VT, FK, and RU assisted with literature search and revised the manuscript. FK drafted 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.


Tandemly repeated satellite DNAs are major components of centromeres and pericentromeric heterochromatin which are crucial chromosomal elements responsible for accurate chromosome segregation. Satellite DNAs also contribute to genome evolution and the speciation process and are important for the maintenance of the entire genome inside the nucleus. In addition, there is increasing evidence for active and tightly regulated transcription of satellite DNAs and for the role of their transcripts in diverse processes. In this review, we focus on recent discoveries related to the regulation of satellite DNA expression and the role of their transcripts, either in heterochromatin establishment and centromere function or in gene expression regulation under various biological contexts. We discuss the role of satellite transcripts in the stress response and environmental adaptation as well as consequences of the dysregulation of satellite DNA expression in cancer and their potential use as cancer biomarkers. Satellite DNAs are tandemly repeated sequences preferentially located in the (peri)centromeric and subtelomeric heterochromatin of many eukaryotes, and they are clustered there in long megabase size arrays . HoweverDrosophila melanogaster, female germline satellite DNA-derived piRNAs are involved in heterochromatin establishment at their own genomic loci and satellite transcription is heterochromatin-dependent [D. melanogaster males, satellite-derived siRNA direct chromatin modification at 1.688 X chromosome satellite repeats, helping the dosage compensation machinery to identify the X chromosome [Tribolium castaneum, the major (peri)centromeric satellite DNA TCAST1 is expressed into piRNAs in the germline and into small interfering RNAs (siRNAs) in somatic cells [in cis at the genomic loci from which they derive (Satellite DNA repeats located in (peri)centromeric heterochromatin are actively transcribed with transcripts being necessary for the formation and maintenance of heterochromatin as well as centromere function ,13,14. Tependent , while iromosome . In otheic cells . TCAST1 y derive . It was T. castaneum, the burst of major TCAST1 satellite transcription occurs during early embryogenesis, coinciding with the initial establishment of constitutive heterochromatin [Aedes aegypti, satellite-derived piRNAs participate in the degradation of transcripts during early embryogenesis [In mammals, RNA Pol II transcribes the pericentromeric satellite DNA repeats into long non-coding RNA in a bidirectional fashion and dsRNAs can be recognized and possibly cleaved by Dicer1, which seems to control the level of satellite transcripts during mitosis ,30. In mhromatin , while iogenesis .The level of transcripts detectable within the active human centromere is low and is in contrast with the higher transcriptional levels of pericentromeric satellites , which aper repeat unit, whereas the density of methylated CpGs is higher at the pericentromeres [Arabidopsis they are significantly methylated [Apart from CENP-A, centromeric and pericentromeric regions, both composed of satellite DNAs, are also characterized by different histone modifications: H3 lysine 4 and lysine 36 methylations (H3K4me1/2 and H3K36me1/2) at the centromere and H3K9me2/3 and H4K20me3 at the pericentromeric heterochromatin ,54, whictromeres ,59. Recetromeres . Also, αtromeres . Plant cthylated ,63. Although it is evident that expression of satellite DNAs is tightly regulated under physiological conditions, under specific conditions and in several biological contexts their expression is significantly changed ,12. HeatT. castaneum (peri)centromeric satellite DNAs, TCAST1 and human α satellite specifically, occurs upon heat stress [T. castaneum satellite DNAs is not induced by HS, which can be related to their genome organization, characterized by preferential location in euchromatin [in trans, targeting homologous regions in euchromatin  conditions protect cells against the toxicity of chemotherapeutic drugs, most prominently the topoisomerase 2 (TOP2) inhibitor etoposide . Recent According to the current results, it is evident that satellite RNA can promote tumor progression by different mechanisms: inducing mutations , affectiSince satellite DNA overexpression occurs in cancer tissues and their transcripts are released into the bloodstream, it is possible to use such circulating satellite RNAs as biomarkers for various types of cancers  Table 1Table 1. Recently, it was shown for patients with metastatic prostate cancer that the increased level of satellite RNA is not only characteristic for cancer, but can be detected in peripheral tissues such as blood cells, serving as a diagnostic marker for metastatic castration-resistant prostate cancer as well as a marker for monitoring the progress of metastatic disease . A mechaBesides satellite RNAs, which are overexpressed in cancer and serve as cancer biomarkers, satellite DNA exhibits change in DNA methylation and copy number variation in different cancers  Table 1Table 1. Identification of epigenetic marks for satellite DNAs located at the centromere, pericentromeric heterochromatin as well as at satellite repeats dispersed within the euchromatin is expected to explain chromatin “compartmentalization” and transcriptional regulation of these sequences. Also, the sequencing and assembly of long arrays of satellite DNAs using new technologies could give insight not only into their structure, organization, dynamics and evolution, but also the regulation of their expression. Further characterization of the transcriptional machinery affecting satellite DNAs and the processing of their transcripts into different forms of small and long RNAs as well as characterization of the proteins associated with them is expected to contribute to the understanding of the physiological role of satellite DNAs and their transcripts under standard conditions and in different diseases. Analysis of RNA modifications of satellite transcripts and their functional significance is an unexplored field of research. In addition, the study of spatial 3D organization of satellite DNAs could show if there is an interaction between heterochromatic and euchromatic satellite DNA repeats, which might influence their expression and genome-wide gene expression regulation. To better understand the role and regulation of satellite DNAs and their transcripts in different pathological processes, the development of new experimental tools for functional studies is needed. Further study of satellite DNAs and RNAs as diagnostic, prognostic or monitoring biomarkers is an area that may lead to novel parameters in diagnosis and treatment strategies.


The formation of subcellular compartments is a characteristic feature of eukaryotic life. Membrane-bound organelles such as mitochondria, peroxisomes, or lipid droplets do however not function as isolated entities; they need to cooperate and to communicate with other subcellular compartments in order to support metabolic pathways and functioning of the cell as a unit. In this regard, organelle cooperation/communication events have been associated with apoptosis, antiviral defence, organelle division and biogenesis, ROS metabolism and signalling, and various metabolic pathways such as breakdown of fatty acids. Coordinated interplay between organelles is often mediated by membrane contact sites (MCS), which bring organelles in close apposition . MCS canKors et al. The authors discuss how the FFAT motif of VAPs allows interaction with over 50 confirmed binding partners and how regulation of these interactions can allow specific control of membrane contact site formation.The ever expanding roles of the VAPs, a group of membrane proteins which play a critical role in connecting the ER to other organelles, are reviewed by Liao et al. summarize the current knowledge on the function and molecular composition of lipid droplet (LD) contacts with mitochondria, lysosomes and the ER. They highlight novel unexpected functions and discuss how LD contact site formation is influenced or regulated by organelle interaction with the cytoskeleton.Ravi et al. explore a novel area of membrane contact site regulation, that of the role of lipid modulators in regulating organelle crosstalk by modifying the lipid composition of membranes. Specifically focusing on Phosphotidylinositol-5-phosphate-4-kinases (PI5P4Ks), which regulate phosphatidylinositol 4,5-bisphosphate  levels, they explain how regulation of lipid phosphorylation on organelle membranes is able to regulate interaction with lipid-interacting proteins.Regulation of membrane contact sites by various different mechanisms is an emerging but still relatively understudied area of research. Islam et al. present new hypotheses for how membrane contact sites could influence the regulation of low density lipoprotein receptor (LDLR) internalisation and degradation. They highlight several contact site proteins that appear to be involved in this process and suggest that more emphasis should be given to understanding how membrane contact sites influence specific events in endocytosis and trafficking.The number of cellular processes, which involve membrane contact sites, is increasingly expanding. In line with, this Sargsyan et al. present an interesting report on the role of calcium in peroxisomes, a topic which has not been extensively explored previously. They discuss the evidence for the presence of calcium in peroxisomes and potential peroxisomal calcium-handling proteins and also speculate on how membrane contact sites with other organelles may facilitated calcium exchange.Bittner et al. review proteins, which are shared by peroxisomes and other organelles. They discuss the mechanisms to achieve dual targeting, their regulation and functional consequences with a major focus on yeast and fungi. They propose that the regulation of dual targeting will further emerge as a major factor to control organelle interplay and communication.Many peroxisomal functions require factors also relevant to other cellular compartments. Wei et al. identified Vps13 as a protein, which is essential for peroxisome formation in yeast mutants with reduced peroxisome-ER contacts. The findings suggest that in the yeast Hansenula polymorpha Vps13 plays a redundant function in lipid transfer from the ER to peroxisomes.How lipids are transferred at membrane contact sites is still not fully understood. Using a mutagenesis strategy, Wu and van der Klei present a first detailed structure-function analysis of Hansenula polymorpha ER protein Pex32, a protein of the Pex23 protein family, which only occur in yeast and filamentous fungi. HpPex32 is required for the formation of peroxisome-ER contact sites. A domain analysis now revealed that the N-terminal transmembrane domain of Pex32 contains ER sorting information, whereas the C-terminal DysF domain is required to concentrate Pex32 at peroxisome-ER contacts and can associate with peroxisomes.Li et al. discuss the impact of peroxisome-derived H2O2 on the regulation of autophagy/pexophagy. They describe our current knowledge on how H2O2 might interfere as a signalling molecule in autophagy regulation by oxidizing thiol groups in autophagy-relevant proteins. The authors propose the concept of a peroxisome-autophagy signalling axis, which might adapt the cellular organelle status to distinct metabolic or stress situations.Van Roermund et al. investigated the metabolic communication between the peroxisomal matrix and the surrounding cytosol. They show that a heterodimer of the yeast peroxisomal ABC transporters Pxa1p and Pxa2p, which transport long chain acyl-CoAs, can as well import cytosolic ATP into peroxisomes. In contrast to the known peroxisomal ATP transporter Ant1p, the ABC transporter heterodimer does not work as an ATP—ADP/AMP antiporter system thereby increasing the flexibility of peroxisomes to increase intraorganellar ATP concentrations. The ATP transporting function is also preserved in the human peroxisomal ABC transporters ABCD1, ABCD2 and ABCD3. Moreover, yeast peroxisomes appear to harbour a third ATP importing system, as the authors further detected that the mitochondrial membrane ATP transporter Aac2p is dually targeted to mitochondria and peroxisomes in Saccharomyces cerevisiae. Thus, yeast peroxisomes appear to utilize three complementing ATP importing systems to adapt intraperoxisomal ATP levels to distinct metabolic states of the cell.The reviews and research articles presented in volume II of this special topic once again demonstrate the impressive breadth of research currently being undertaken to understand the molecular mechanisms and physiological significance of organelle interactions and cooperation.


Aphasia is an acquired central disorder of language that affects a person’s ability to understand and/or produce spoken and written language, caused by lesions situated usually in the dominant (left) cerebral hemisphere. On one hand aphasia has a prevalence of 25–30% in acute ischemic stroke, especially in arterial infarcts. On the other hand, cerebral venous and dural sinuses thrombosis (CVT) remains a less common and underdiagnosed cause of ischemic stroke . Aphasia has been observed in almost 20% of patients who suffered CVT. The presence of aphasia is considered a negative predictive factor in patients with stroke, severe language disorders corresponding to arduous recovery. Taking into consideration data from the literature, aphasia is also considered a predictive factor for patients with CVT; its absences, together with the absence of worsening after admission, are determinants of complete recovery after CVT. This review has as the principal role of gathering current information from the literature (PubMed database 2012–2022) regarding the clinical features of aphasic syndromes and its incidence in patients with CVT. The main conclusion of this review was that aphasic syndromes are not usually the consequence of isolated thrombosis of dural sinuses or cerebral veins thrombosis. The most frequent form of CVT that determines aphasia is represented by the left transverse sinus thrombosis associated with a posterior left temporal lesion , followed by the superior sagittal sinus thrombosis associated with a left frontal lesion . Only a few cases are presenting isolated cortical veins thrombosis and left thalamus lesions due to deep cerebral vein thrombosis. We also concluded that the most important demographic factor was the gender of the patients, women being more affected than men, due to their postpartum condition. On one hand, aphasia, which is an acquired central disorder of language, is one of the most common focal neurologic deficits observed in CVT, being noticed in 19–24% of cases . Unlike CVT is a special category of cerebrovascular disease, encountered especially in children and younger adults rather than in arterial ischemic strokes , mostly women . Early acquaintance of CVT’s clinical features (including language disturbances) may improve the prognosis of this potentially fatal pathology.The term of aphasia includes the inability of a person to understand and/or produce spoken language, sometimes also accompanied by the incapacity of reading  and writing (agraphia). It is always the consequence of an acquired central disorder, meaning that it occurs after the language has already been developed. Aphasia should be differentiated from a peripheral disorder of language (such as weakness or discoordination of phonatory musculature) that may easily simulate aphasia .The organization of language networks has not been yet fully elucidated, despite the technological advances made in past years. Using complex techniques, especially functional neuroimaging, scientists have proven that language production is the result of neural activation from a vast network scattered in different structures of the brain: the cortex, the basal ganglia, the cerebellum and even the brainstem. This is the reason a disruption of this network determines spectrum of clinical signs. Functional neuroimaging studies suggested that the core of language operates in the left perisylvian regions in most individuals (95% right-handed and 75% left-handed), and there are two principal pathways in the language network: the dorsal fronto-parietal pathway (which is responsible for articulatory and syntactic tasks) and the ventral temporal pathway  ,9:Broca’s area—the posterior part of the left third frontal gyrus (F3)—Brodmann areas 44 and 45;Left insula’s cortex and the underlying white matter;Left Rolandic operculum—the lower part of the motor area (Fa);Left premotor and prefrontal areas (forward and superior of Broca’s area);The supplementary motor area;Anterior areas  are represented by:Wernicke’s area: the posterior part of the first two temporal gyri-T1/T2 (Brodmann area 22);The inferior parietal lobes: the angular gyrus (Brodmann area 39), and the supramarginal gyrus (Brodmann area 40);The anterior part of the temporal lobe.Posterior areas  are represented by:The most important determining factors of aphasias are the etiology, the site, and the magnitude of the causative lesion/lesions . Age andBroca’s aphasia (10–15%);Wernicke’s aphasia (15%);Conduction aphasia (15%);Transcortical motor aphasia (15–20%);Transcortical sensory aphasia;Mixed transcortical aphasia.Transcortical aphasias:Global aphasias (24–38%);Anomic plus aphasias (20%);Atypical aphasias: mixed aphasias, thalamic aphasias, and capsulo-striatal aphasias (10%).Taking into consideration the clinical features of aphasia, there are seven types of aphasic syndromes ,10,11,12More than 10% of aphasic syndromes remain unclassifiable, particularly in patients with ischemic stroke history.Regarding the etiology of aphasic syndromes, there are various cerebral lesions  that can disturb language networks usually situated in right-handed individuals in the dominant hemisphere  . The mosWe achieved a literature search in the PubMed database [cerebral phlebothrombosis aphasia-Search Results-PubMed (nih.gov)], from 2012 to 2022 (in the present), using the following search terms: “aphasia” and “cerebral phlebothrombosis”. Our main objective was to identify the incidence of aphasic syndrome, the main characteristics of those syndromes and correlation between clinical picture and localization of cerebral veins or dural sinuses thrombosis. After identifying the articles, we reviewed the abstracts of the papers and made a preliminary evaluation of their eligibility. After this stage, all potentially eligible papers were accessed and read. We included only those papers which met the eligibility criteria and were relevant to our review .(1)Presence of language disturbances  and description of aphasia ‘s type or characteristics;(2)Case-report studies published between 2012–2022;(3)Adult human studies;(4)Articles written in English;(5)The patients included in the study were diagnosed based on an imaging examination: computed tomography (CT), computed tomography venography (CTV), magnetic resonance imaging (MRI), magnetic resonance venography (MRV), intra-arterial angiography ;(6)Detailed, reliable medical history, physical examination, results of laboratory and imaging examinations were required.We applied the following eligibility/including criteria:The principal exclusion criteria were the absence of aphasia or lack of information regarding aphasia (N = 21), not being a case-report study (N = 15), absence of CVT and presence of other causes of stroke (N = 5).After carefully analyzing all these studies , we noteThe first is that the interest for characterizing the distinctive features of aphasic syndromes is extremely poor. The authors usually specify only the term of “dysphasia”, “aphasia” or “language disturbances”, without fitting into a certain aphasia type. This fact explains the difficult identification of eligible articles for our study and consecutively their small number.n = 9/10), followed by the superior sagittal sinus thrombosis associated with a left frontal lesion , with a percentage of 40% (n = 4/10). Only 20% of cases were presenting isolated cortical veins thrombosis (n = 2/10) and 10% (n = 1/10) left thalamus lesions due to deep cerebral vein thrombosis. Our results are comparable to recent data from the literature. According to International Study on Cerebral Vein and Dural Sinus Thrombosis, aphasia is commonly the consequence of left transverse sinus thrombosis, followed by the superior sagittal sinus thrombosis [The second result is that the most frequent site of CVT that determines aphasia is represented by the left transverse sinus thrombosis associated with a posterior left temporal lesion , with a percentage of 90%  and the global/mixed aphasia (percentage of 30%). Only one case of fluent aphasia  was described in the case-report studies included in our research.The third result is that the most frequent type of aphasias found in patients with CVT are: the non-fluent type (Broca’s aphasia and anomic aphasia) with a percentage of 50% (n = 7/10), due to their post-partum and post abortion condition.The fourth result is that young female patients , oncological cases and severe medical conditions  are having a higher risk in developing CVT. From our statistical data, the mean age of female patients was 44.42 years , meanwhile the mean age of male patients was 48.87 years . Regarding the sex incidence, female patients are leaders in CVT  thru the arachnoid villi. As a result, intracranial pressure increases , usually in association with superior sagittal sinus (SSS) thrombosis. All these pathophysiological altered mechanisms lead to typical focal neurologic signs and symptoms, depending on the territory of the brain in which the venous drainage is altered, the rapidity of the thrombosis process , and the magnitude of brain damage. Superior sagittal sinus (62–80%) and lateral sinus 38–86%) are the most frequently involved sites for thrombosis [8–86% areRegarding the focal neurologic deficits, specific clinical symptomatology and neurological signs can suggest which area of the brain is damaged. However, the topographic clinical diagnosis of CVT is not as well-defined as in arterial occlusion, frequently being misleading. This characteristic is the consequence of concomitant multiple cerebral veins and dural sinuses thrombosis (more than two-thirds of cases), the numerous anatomic variants of some dural sinuses and some cerebral veins, and the existence of venous collateral circulation ,25.When aphasic syndromes are the consequence of dural sinus/sinuses thrombosis or cerebral veins thrombosis, establishing the exact site of lesions might be a challenge. However, the clinical aspects of aphasic syndromes may vary depending on the concomitant occlusion of other dural sinuses or cerebral veins.Superior Sagittal Sinus (SSS) ThrombosisIn SSS thrombosis, aphasia is the result of left fronto-parietal hemispheric lesions, secondary to progression of thrombosis to the tributaries bilateral cortical veins ,16,25,262.Lateral Sinus (LS) ThrombosisLateral sinus contains two parts: the transverse sinus and the sigmoid sinus, respectively. Regarding the left transverse sinus thrombosis, it is well known that the association of adjacent temporal veins occlusion is the one that produces fluent Wernicke aphasia (40%) ,25.Another specific characteristic of this dural sinuses is that the left LS is often hypoplasic; due to this characteristic, after right LS thrombosis, a pseudotumor syndrome is installed, causing bilateral venous drainage deficiency, affecting contralateral structures, especially the inferior part of temporal lobe and cerebellum .n = 7/10).In many cases, thrombosis of lateral sinus spreads to other sinuses  and veins ,13,26,273.Isolated Cortical Veins ThrombosisWithout association with dural sinus thrombosis, isolated thrombosis of cortical veins is considered a rare affection (2%) ,25. A pl4.Deep Cerebral Veins ThrombosisDeep venous system thrombosis can be suspected in patients with altered state of consciousness  and motor deficits  ,26. DeepOver the last few years, several case reports have brought to attention the small incidence of aphasia in the clinical picture of CVT. Unfortunately, few of those studies made a precise evaluation of the aphasic syndrome and a clinical-imagistic correlation.Alexander Engelmann and his collaborators  publisheGiovana Ennis and her team  brought Another recent study conducted by Ayele et al. , reporteIn 2014, Tuncel and his collaborators  reportedIn 2021, Jianu et al. , handledSearching into the specialized literature, we observed a small number of observational studies which followed the clinical features of aphasic patience with CVT. The VENOST study  is the l-aphasia is commonly the consequence of concomitant left transverse sinus thrombosis with left cortical temporal veins, followed by the superior sagittal sinus thrombosis with extension into tributary fronto-parietal cortical veins;-depending on the site and the size of the brain damage , the most frequent types of aphasias are: non-fluent aphasias-Broca’s aphasia and anomic plus aphasia (if the lesions are situated in the anterior language areas), fluent aphasias-Wernicke’s aphasia, transcortical sensory aphasia (if the lesions are situated in the posterior language areas), mixed or global aphasias (in the cases of larger lesions);-in LS sinus thrombosis  associated with left cortical temporal veins thrombosis, the most common type of aphasia is Wernicke’s aphasia (40%) ;-in many cases, the LS thrombosis spreads to the SSS, symptoms and signs of SSS thrombosis depending on the involvement of cerebral veins and other dural sinuses. The most often involved are the superior cerebral veins  which empty into the SSS. If the thrombosis spreads to the deep veins system, altering the Ascending Reticular Activating System (ARAS), awakening alteration may occur;-the clinical picture depends on the location and the dimensions of cerebral lesion: extensive lesions can determine global aphasia, or mixed transcortical aphasias; meanwhile smaller lesions might determine Wernicke’s aphasia, transcortical sensory aphasia, Broca aphasia or anomic aphasia.Analyzing all this data, we can easily observe that it is hard to make a statistically significant hypothesis regarding the incidence of different types of aphasic syndromes in CVT, taking into consideration that the diagnosis of a certain type of aphasia requires specific assessments, using approved tests, while the patient is cooperating . However, some conclusions are clear from this study:Our small study managed to highlight the small amount of information that has been gathered until now regarding the characteristics of aphasic syndromes in CVT, and this might be considered a great limitation. Unfortunately, the small number of cases described in the literature made difficult our intention to demonstrate a certain pattern of language disturbances, using the localization of the CVT. Mostly, the greatest impediment was the fact that few authors focused their attention on the clinical characteristics of aphasic syndromes, analyzing and describing the clinical aspects of language disturbances in an incomplete manner. However, this might be the beginning of a new research theme in this rare and underdiagnosed condition. We aim to continue our study and to gather further information over the next years to support us in outlining a clinical template that will guide clinicians in establishing the diagnosis of CVT much faster, more easily, and more accurately.In conclusion, the early identification of aphasia in CVT can help us to provide a better prognosis of patients and the site of the lesion and guide us in the differential diagnosis of this cerebro-vascular pathology. Although aphasia is a rare syndrome in a rare disease (CVT), we should look toward new clinical data regarding this issue, insisting over the examination of the patients’ language and trying to determine whether, the presence of aphasia might be a severity marker as it is considered in stroke. We should carefully look after patients with associated conditions that have fully demonstrated their potentially prothrombotic characteristic, especially in young women (postpartum/post abortion) and patients with inherited thrombophilia.What is worth underlying is that aphasic syndromes are not the consequence of thrombosis of a specific, isolated dural sinus, but the result of thrombosis extension to cortical veins or other dural sinuses . Venous infarction in strategic language areas is also the main determining cause of aphasic syndromes.


Birefringence was investigated in unstained tendon sections using high-performance polarization microscopy and image analysis. LD was studied under polarized light in tendon sections stained with the dichroic dyes Ponceau SS and toluidine blue at pH 4.0 to evaluate the orientation of proteins and acid glycosaminoglycans (GAG) macromolecules, respectively. Structural remodeling characterized by the reduction in the macromolecular orientation, aggregation and alignment of collagen bundles, based on decreased average gray values concerned with birefringence intensity, LD and morphological changes, was detected especially in the tendon explants cultivated on the plastic substrate. These changes may have facilitated cell migration from the lateral regions of the explants to the substrates, an event that was observed earlier and more intensely upon tissue cultivation on the plastic substrate. The axial alignment of the migrating cells relative to the explant, which occurred with increased cultivation times, may be due to the mechanosensitive nature of the tenocytes. Collagen fibers possibly played a role as a signal source to cells, a hypothesis that requires further investigation, including studies on the dynamics of cell membrane receptors and cytoskeletal organization, and collagen shearing electrical properties.Collagenous tissues exhibit anisotropic optical properties such as birefringence and linear dichroism (LD) as a result of their structurally oriented supraorganization from the nanometer level to the collagen bundle scale. Changes in macromolecular order and in aggregational states can be evaluated in tendon collagen bundles using polarization microscopy. Because there are no reports on the status of the macromolecular organization in tendon explants, the objective of this work was to evaluate the birefringence and LD characteristics of collagen bundles in rat calcaneal tendons cultivated in vitro on substrates that differ in their mechanical stiffness (plastic vs. glass) while accompanying the expected occurrence of cell migration from these structures. Tendon explants from adult male Wistar rats were cultivated for 8 and 12 days on borosilicate glass coverslips ( Tendons are a specialized group of dense connective tissue that anchors muscles to bones, provides stability, and transmits muscle forces to bones and absorbs impacts, thus enabling the various extension, flexion, and rotation movements of joints ,2,3,4,5.In terms of composition, tendons are complex structures that are rich in extracellular matrix (ECM) and water that corresponds to 55–70% of the tissue mass . The ECMBirefringence brightness levels revealed by macromolecularly oriented biological structures that were examined using a polarizing microscope are the result of the sum of intrinsic or crystalline birefringence (Bi) and form or textural birefringence (Bf) phenomena. Bi is defined by Bennett  as “anisAmong the types of light microscopy, polarization microscopy has thus been extensively used to investigate the submicroscopic structural characteristics and orientation of molecular components of cells or tissues . Even ifHigh-performance polarization microscopes are built with every optical component free of strain, permitting that any kind of optical compensator  be used, Wollaston prisms be inserted, differential interference microscopy be undertaken, and image analysis procedures be performed .ǁ − A⊥, where Aǁ and A⊥ are the light absorptions exhibited when the long fiber axes of oriented molecular structures are arranged parallel and perpendicular, respectively, to the plane of incidence of polarized light. Because LD in visible light is not usual in biological objects, to study the orientations of collagen and GAG components of collagen fibers using LD properties, it is necessary to stain them with dichroic dyes that have an affinity for proteins  [Linear dichroism (LD) is the selective absorption of polarized light that results from the molecular orientation of the analyzed substrate relative to the axis of the incident light. It can be expressed mathematically as LD = Aceau SS) ,20 or GAceau SS) ,21. Depeceau SS) .Birefringence and linear dichroism studies have contributed to determining the changes in the macromolecular organizations of tendon collagen bundles during tendon repair ,27,28,29Tendon explants have been recognized as a valuable tool to investigate mechanisms associated with tendon homeostasis and pathophysiology  and as sIn addition to their structural functions, tendon collagen bundles interact with cells in highly regulated processes, allowing them to respond to stimuli in specific ways by acting as cellular signal transducers ,40,41,42Tenocytes are specialized mechanosensitive cells that are organized in interconnected networks and are surrounded by small amounts of specialized matrix with their large cytoplasmic extensions connecting adjacent cells along tendons . These cThe object of this work, by using optical anisotropy methods, was to investigate how the macromolecularly oriented organization of adult tendon collagen bundles responds to tissue cultivation in vitro and whether it is affected by explant adhesion substrates with different states of mechanical stiffness (plastic vs. glass). This investigation also aimed to detect whether cell migration in tendons cultivated in vitro is associated with changes in the oriented organizations of tendon fibers and responds differentially under tissue cultivation on glass and plastic substrates. Considering that the methodology used is sufficiently sensitive to detect the submicroscopic structure and orientation of the macromolecular components of the tendon collagen fibers, the results obtained in this work will potentialize further studies on tendon explants obtained from newborn and aged animals where the organization and composition of the collagen bundles and the cellularity abundance are known to differ in vivo ,30. TendRattus norvegicus albinus) (n = 5) were provided by the Multidisciplinary Center of Biological Investigation (CEMIB) at the University of Campinas . The animals were reared under standard conditions, given water ad libitum and fed extruded chow . Animal care and use protocols were approved by the Ethics Committee for Animal Use at the University of Campinas (registration no. 5064-1/2018) and followed the guidelines established by the Brazilian National Council for the Control of Animal Experimentation (CONCEA).Adult male Wistar rats (cov) (n = 3) or placed in direct contact with the surfaces of cultures of 35 mm ø nonpyrogenic polystyrene plastic dishes (pla) (n = 4) for 8 and 12 days in DMEM/HAMF12 (v/v)  medium supplemented with 10% bovine fetal serum and penicillin/streptomycin  at 37 °C, 95% humidity and 5% CO2. The media were changed every 72 h. Tissues were maintained in 4 mL of medium per dish. Tendons not cultivated in vitro were used as controls (n = 3). To improve the adhesion of the tendon explants of the pla group to the dish surfaces, they were incubated in an absence of medium for 60 min in a cell culture oven at 37 °C prior to beginning the proper culture procedures. All tendon explants were maintained without no longitudinal or compressive forces applied to the tissues.Calcaneal tendons isolated from these rats were cultivated under sterile conditions on 24 mm square colorless borosilicate glass coverslips , and cut to a thickness of 8 µm with a Microm HM 315 microtome .Birefringence analysis was performed in tendon sections that were immersed in deionized water , and the maximum brilliance intensity corresponds to the sum of the Bi and Bf phenomena ,12,15. D5 µm2; the number of average gray measurements was between 206 and 297, which was sufficient to cover the possible variations in anisotropy.Because the birefringence brilliance intensities observed in tendon sections were distributed unevenly along the tendon axes and large areas of images had to be analyzed using image analysis, the total area of images obtained after birefringence compensation was segmented into smaller subareas . The sofA polarizing microscope (Olympus BMX-51) equipped with two Wollaston prisms was used for DIC-PLM analysis. Optical path differences were detected by displacing the prism positioned under the analyzer and comparing the resulting interference colors with those on the Michel-Lévy color scale .3+ groups. If the protein -NH³+ groups available to dye binding occur in the substrate with an oriented distribution as in the case of collagenic proteins, the stained substrate will exhibit linear dichroism because the dye molecules attached to the substrate will acquire an ordered stereo arrangement and the oriented dye molecules are susceptible to exhibit a selective absorption of polarized light [Sections 8 µm thick from tendons fixed in 4% paraformaldehyde were stained for 3 min with a 0.025% solution of Ponceau SS  in 3% acetic acid prepared in bidistilled water, followed by rinsing in 3% acetic acid for 2 min and then placed in distilled water to investigate the LD properties of tendon proteins . Ponceaued light . The inted light .− and COO− groups like those present in GAGs. If the available binding sites in the stained substrate are positioned with an oriented distribution, the stereoarrangement of the dye molecules will also be oriented and the phenomenon of linear dichroism will be exhibited using polarization microscopy, thus revealing the status of molecular orientation of the stained substrate [Sections taken from tendons were also fixed in paraformaldehyde and stained with a 0.025% TB (Merck) solution in McIlvaine buffer at pH 4.0 for 15 min and then rinsed in McIlvaine buffer at pH 4.0 and in a stabilizer solution of 4% ammonium molybdate aqueous solution for 4 min to investigate LD in tendon GAGs . The pH ubstrate ,13,14,21ǁ) and perpendicular (A⊥) to the plane of the PPL.The stained preparations were examined under polarized light using an AxioPhot microscope  (Ponceau SS-stained sections) or using an Olympus BMX-51 high-performance polarization microscope (TB-stained sections), both equipped with 40× objectives. The long axis of the tendons was positioned parallel (AThe tenocyte distributions were determined during a 12-day period in tendons cultivated in vitro on plastic and glass surfaces. Examinations were performed using an Olympus IX50 inverted microscope (Olympus America Center), and images were captured using Motic Image Plus 2.0 software .® software . Kruskal–Wallis and Mann–Whitney tests were used to assess statistical significance when comparing GA values. p < 0.05 was considered the critical level to reject the null hypothesis.Calculations and statistical analyses were performed using Minitab 14Typical birefringence brilliance intensities were detected when the collagen bundles of the control  were intense in Ponceau SS-stained collagen fibers of the control tendons (LD phenomena with positive signs (A⊥ > Aǁ) were detected in TB-stained collagen bundles of the control and cultivated tendons, although less intense colors were observed in the cultivated tissues (Typical LD phenomena with negative signs (A tissues a–f. Diff tissues a–f.While tenocyte migration was evident immediately after a 5-day culture period in tendons cultivated on plastic dishes a,b, migrCell migration occurred primarily from the lateral zones of tendons , althougInitially, cells that migrated to the surfaces of the plastic dishes appeared to be distributed without a preferential direction a,b. WithThe optical anisotropic birefringence and LD results obtained demonstrated structural remodeling that was characterized by changes in macromolecular orientations and aggregational states of the collagen bundles in rat tendons subjected to tissue culture for 8 and 12 days, especially upon cultivation on a plastic substrate. The occurrence of the optical anisotropic phenomenon of birefringence has been widely reported for tendon collagen bundles based on visual observations or quantitatively evaluated through measurements of optical retardations . The decCavia porcellus L. [Determinations of optical anisotropy properties have already enabled the detection of changes in the stereo arrangement of the macromolecular components of collagen bundles in tendons under different physiological states, such as in detached or surgically implanted tendons of ellus L. , during ellus L. ,25,28, iellus L. , in rat ellus L. , after tellus L. , and in ellus L. . Valdetaellus L. , who useUpdated studies reinforce the applicability of polarization microscopy in determining molecular order in tendon structures. The evaluation of birefringence expressed as average gray values has allowed the detection of improvement in the state of aggregation and organization of tendon collagen bundles in a study using low-level laser and adipose-derived stem cells . BirefriAn advantage of the present detection of changes in birefringence intensity using image analysis was that the uneven distribution of the gray values corresponding to compensated birefringence across the tendon section could be evaluated quickly and across a large image area of the preparations. The use of the polarization microscopy methods described in this study has a limitation which is a requirement of experimental expertise from the microscopist. The added value of these methods is that images containing informative algorithms fundamental for the interpretation of the results can be obtained.Although there were reductions in the birefringence GA values of collagen bundles and decreased LD values for the Ponceau SS-reactive proteins in tendons cultivated directly on glass coverslips or plastic dishes, tissue remodeling was more visually evident, and cell migration occurred earlier and more intensely in samples that were incubated in an oven and then cultivated on the surfaces of plastic dishes. Substrate stiffness differences upon which tendon cultivations occurred thus differentially affected the rate of the altered stereo arrangement process of tendon structures and permitted cell motility and migration toward the culture adhesion substrate with increasing culture time . The decPreferential migration of tenocytes to lateral regions of rat tendon explants is consistent with reports showing that cells originating from the surfaces of equine digital flexor tendon explants migrated faster and exhibited higher replication rates than cells originating from the inner regions of these tendons . MigratiMechanical interactions between cells and collagen fibers are known to affect tissue architecture ,43. AccoThe cell migration phenomenon in rat tendons cultivated in vitro demonstrates the performance of the structural components of tendons, with special emphasis on collagen, in its role as a transducer of electrical signals and potentials. As previously reported, the superstructures of collagen bundles endow them with anisotropic, piezoelectric, and pyroelectric optical properties, in addition to electric transmission potentials, that extend from molecular to nanometric organizational levels. Chirality, rotational symmetry, and liquid crystal twisted grain boundaries are necessary conditions for the occurrence of such properties . MechanoThere are reports describing increased numbers of tenocytes in lesion areas during late-stage healing in rat tendons, where these cells proliferate and become responsible for the abundant deposition of ECM ,65,66. IStructural remodeling involving reductions in crystallinity levels, which were evaluated based on the changes in optical anisotropic characteristics such as birefringence and linear dichroism occurs in rat calcaneal tendon explants cultivated in vitro for 8 and 12 days on plastic and glass substrates. These changes affect the aggregation states and macromolecular orientations of components in tendon collagen bundles, such as collagen and non-collagen structural glycoproteins and GAG, and are accompanied by tenocyte migration to culture substrates. Glass substrates appear to delay the structural rearrangement process of collagen fibers and of tenocyte migration, possibly because of their stiffness. The plastic substrate used for calcaneal tendon explant adhesion reveals more suitable than glass to detect changes in the stereo arrangement of collagen bundles and facilitation of cell migration during tissue culture. Cell migration and alignment in rows parallel to the long axes of the tendons cultivated on plastic substrates are possibly due to the mechanosensitive nature of tenocytes. Further studies involving digestion of GAG components by specific enzymes would certainly reveal the level of participation of these macromolecules on the changes in stereo arrangement of collagen bundles and cell migration detected during tendon explant culture.


An enhanced primary care team model was implemented to provide proactive, longitudinal care to patients with diabetes, grounded in close partnership between primary care providers (PCPs), nurses, and Medication Management Services (MMS) pharmacists. The purpose of this study is to evaluate the impact of the MMS pharmacist involvement in the enhanced primary care model for patients with diabetes.This retrospective cohort study compared the quality of diabetes care between patients referred to a pharmacist and propensity score matched controls who were not. Eligible patients were adults (age 18 to 75 years) enrolled in the enhanced primary care team process who did not meet at least one of four diabetes quality indicators at 13 Mayo Clinic Rochester primary care practice locations. The intervention examined was asynchronous e-consults by pharmacists affiliated with the primary care practice.The primary outcome was change in the proportion of patients meeting the composite of four diabetes treatment goals (D4), including hemoglobin A1c (HbA1c) control, blood pressure control, aspirin use, and statin use at six months from enrollment among patients who received pharmacist intervention compared to matched patients who did not. Secondary outcomes were each of the D4 goal individually.The proportion of patients meeting the D4 increased with pharmacist e-consults (N = 85) compared to matched controls with no review (N = 170) . The change in patients meeting treatment goals of HbA1c , blood pressure , aspirin use , and statin use  all increased with pharmacist e-consults.Pharmacist engagement in the enhanced primary care team improved diabetes management. This supports the inclusion and utilization of pharmacists in multidisciplinary efforts to improve diabetes care. Diabetes is one of the leading causes of morbidity and death in the United States, affecting 37.3 million Americans in 2019 . To helpTo improve the care provided for this complex and multifaceted condition and to address the increasing demand for health services stemming from increased prevalence of diabetes and comorbidity and a diminishing primary care workforce, innovative care models utilizing pharmacist consultative services offer a promising optimization of patient care . While aWe have previously reported the development and implementation of an enhanced primary care team model comprised of care team registered nurses (RNs) and MMS pharmacists designed to improve the quality of diabetes care provided to patients . The MMSWhile efficient and likely effective in principle, this approach to interdisciplinary diabetes care management has not been previously evaluated for feasibility or effectiveness. Thus, to address this knowledge gap, we examined the feasibility and impact of pharmacist engagement on diabetes care quality, successfully hypothesizing that utilizing the MMS pharmacist team for treatment recommendations would improve treatment goals of diabetes care. Determining the utility of this novel engagement between pharmacists and the care team may help inform best practices for this process and the development of other multidisciplinary models of diabetes and other chronic disease management.This retrospective cohort study compared outcomes among adult patients with diabetes whose charts were asynchronously reviewed through an e-consult by a MMS pharmacist as part of the enhanced primary care team diabetes management model, and for whom care recommendations were provided, with outcomes among matched patients with diabetes who were also managed by the enhanced primary care team model but did not receive a pharmacist review. The study was classified as quality improvement and deemed exempt by the Mayo Clinic Institutional Review Board. All included patients provided authorization to use data from the electronic health record (EHR).Mayo Clinic is an integrated healthcare delivery system based in Rochester, MN, U.S.A that delivers the full continuum of care to local, regional, national, and international patients. Thirteen primary care practices located in Olmsted (12) and Dodge (1) counties deliver comprehensive primary care to local patients, Mayo Clinic employees, and their dependents. The practices are located in urban, suburban, and rural areas with both Family Medicine and Internal Medicine practices. All patients are paneled to a staff physician, nurse practitioner (NPs), physician assistant (PAs), resident, or fellow.A commonly used indicator of diabetes care quality is the D5, developed and used by the Minnesota Community Measurement Program (MNCM), an organization that delivers data to healthcare payer and provider members to illustrate performance on quality and cost measures . The D5 The study population was comprised of patients meeting eligibility criteria for MNCM D5 reporting. Eligible patients were adults aged ≥18 to ≤75 years old with a diagnosis of type 1 or type 2 diabetes who are paneled to a PCP at one of the Mayo Clinic Rochester primary care clinic locations. Patients were excluded from the study if they were incarcerated, pregnant, enrolled in hospice or receiving palliative care, a permanent nursing home resident, died during the measurement period, declined research authorization, or had not had a PCP visit during the measurement period. Of patients enrolled in the enhanced diabetes care process not meeting D5 treatment goals, those with a MMS pharmacist e-consult and care recommendations documented in the EHR during our study period were compared with those participants without documented pharmacist involvement. Patients who entered the care model between May 2019 and February 2020 were included.The enhanced primary care team model was implemented across all 13 primary care practices of Mayo Clinic Rochester in May 2019 . BrieflyOur primary outcome was the change in the proportion of patients meeting the D4 treatment goals, which is comprised of HbA1c <8.0%, blood pressure <140/90mmHg, aspirin use for secondary prevention of ischemic vascular disease, and cholesterol lowering through statin use as recommended, between the intervention and control groups at six months from the index date. The tobacco use component of the D5 was not included due to an EHR error identified during the study period that incorrectly reported tobacco use. Our secondary outcomes included change in the proportion of patients meeting each of the four D4 components at six months from the index date. Additionally, we examined the change in the proportion of patients with type 2 diabetes treated with specific classes of evidence-based medications at four weeks from the index date. The 4-week time point was chosen to more reliably connect any medication change to the e-consultation. These were multi-dose insulin (MDI), sodium-glucose co-transporter 2 inhibitors (SGLT2i), glucagon-like peptide-1 receptor agonists (GLP-1 RA), angiotensin-converting enzyme inhibitors (ACEi), and angiotensin receptor blockers (ARB).We used propensity score (PS) matching to reduce the imbalance of measured baseline characteristics between patients with and without a pharmacist review in a 1:2 manner. The PS model was designed to capture control patients who should have received a pharmacist review but did not. Parameters included in the matching were age, race, diabetes type, baseline HbA1c, baseline blood pressure, number of provider visits in the past year with a visit diagnosis of diabetes, LDL-C level, endocrinology visits in the past year, and insulin use, and was estimated by a multivariable logistic regression model. Patients were then matched on the logit of the PS (± 0.2 standard deviation of the logit of the PS). Chi-square tests were used to compare categorical data, including individual and composite D4 treatment goals between groups. Among those not meeting given individual goals, Wilcoxon rank sum tests were used to compare HbA1c, systolic blood pressure, and diastolic blood pressure values between groups. All analyses were conducted using SAS version 9.4 software .Our study compared 85 patients with diabetes enrolled in the enhanced primary care team model who received pharmacist e-consultation to 170 matched patients who did not. Baseline characteristics of the two groups are summarized in The change in the proportion of patients meeting the D4 was greater in the pharmacist intervention group compared to the usual care group after 6 months  . The chaOverall, there were no significant differences between groups in the change from baseline to four weeks from the index date in the proportion of patients taking multi-dose insulin , SGLT2i , or ACE-Is/ARBs . However, there was a larger increase in the percent of patients taking GLP-1 RA from baseline to 4 weeks in the pharmacist review group compared to the non-pharmacist group .In this post-hoc evaluation of an innovative multidisciplinary enhanced primary care model for the management of diabetes implemented across 13 primary care practices in the U.S. Midwest, engaging a pharmacist for treatment recommendations via asynchronous e-consultation significantly improved attainment of diabetes treatment goals over a six-month period. When examined individually, each of the diabetes treatment goals, including HbA1c, blood pressure, aspirin use, and statin use improved with pharmacists’ recommendations on the treatment plan. These data support the integration of pharmacists into the primary care team and demonstrate their ability to efficiently and effectively support chronic disease management through asynchronous e-consultation.While several prior studies have demonstrated the effectiveness of pharmacist consultations with respect to improving diabetes management, our work extends this benefit to asynchronous e-consultation recommendations organized by primary care nurses, a more efficient alternative to patient consultation that can be scaled and generalized to other settings and populations . In contAs primary care practices deliver the majority of care to patients with diabetes across the United States, therapeutic inertia, defined as “a lack of timely adjustment to therapy when a patient’s treatment goals are not met,” is a growing challenge that hinders optimal diabetes management . ComprehThe enhanced primary care model illustrates that not all patients with diabetes not meeting care goals require intensive, longitudinal interventions by pharmacists. Pharmacists are a more finite resource than nurses, and the e-consultation process allows pharmacists to improve the care of more patients and for patients located in different physical locations. Longitudinal pharmacist care can then be reserved for more complex patients, such as those on MDI regimens or those not making progress through the enhanced care model. Additionally, pharmacists played a vital role in supporting care team RNs by educating them about potential treatment options to discuss with the PCPs and patients, ultimately empowering and enabling them to take an increasingly active role in patient education and chronic disease management. Pharmacists provide valuable insight and mitigation strategies on medication barriers such as cost, side effects, and administration which can help nurses engage patients in informed shared decision making and ensure that a treatment plan is finalized in one patient conversation, without back-and-forth messaging with the PCP. Pharmacists were also embedded within the primary care clinic with established relationships within the care team, which assists in buy-in to seeking advice and implementing the recommendations. Additionally, this model allows for bi-directional learning between nurses and pharmacists through direct interactions, leading to improved collaboration among primary care team members.When examining the use of specific medication classes between the groups, we found that only GLP-1RA prescribing increased in the pharmacist e-consult group. A larger sample size may be needed to detect differences in multi-dose insulin or SGLT2i use between groups. The high baseline rate of ACEi/ARB use, nurses and PCPs familiarity with these established medication classes, and an existing EHR prompt to prescribe ACEi/ARB to patients with diabetes and hypertension may explain why these rates did not differ between the two groups.Strengths of the study include the assessment of multiple clinics in urban, suburban, and rural areas and within both Family Medicine and Internal Medicine practices, which increases generalizability. The pharmacist intervention is generalizable and feasible as it can be completed virtually and is less resource intensive compared to more traditional pharmacist models.Limitations of our study include the retrospective design which may not account for all potential confounders and does not establish causality, though we did utilize a propensity score matching approach to balance groups on measured confounders. There are approximately 7,600 eligible patients with diabetes aged 18–75 years within our healthcare setting, and this pilot evaluated a smaller sample size for patients who received pharmacist e-consultation. Additionally, we could not evaluate the D5 tobacco-free treatment goal due to a known EHR reporting error. This resulted in some patients meeting D4 at baseline. Less patients met the D4 at baseline in the pharmacist group which was likely due to limited pharmacist e-consult requests for cessation treatment recommendations. We did not include the aspirin goal in our PS matching due to high rates of attainment in the overall population, though fewer patients met this goal at baseline in the pharmacist group. The D5 composite measure is comprised of surrogate outcomes and process measures rather than hard clinical outcomes, such as cardiovascular events, and the long-term impact of pharmacist engagement on patient-centered hard outcomes will need to be determined as this care process continues to be implemented in the practice. Finally, the study was conducted at 13 practices within the same health system. Expansion to other health systems and sies is needed to fully establish the feasibility and effectiveness of the pharmacist e-consultation model embedded within primary care.Future studies should examine the overall enhanced primary care team model and the pharmacists’ role within it specifically for patients at highest risk for suboptimal diabetes management and diabetes-related complications. This includes racial and ethnic minoritized populations, rural residents, low-income individuals, and patients with limited English proficiency. Further, it will be important to assess the long-term impact of this model on microvascular and macrovascular complications of diabetes, patient satisfaction with care, and clinician burnout.The enhanced primary care team model that leverages MMS pharmacists through asynchronous e-consultation is a novel multidisciplinary effort to improve the quality and efficiency of care for patients with diabetes. MMS pharmacists embedded in primary care teams have a high level of expertise in chronic disease management and are often a limited resource. Therefore, it is important that pharmacists continue to find innovative ways to positively impact the care of more patients. This study demonstrates a novel approach for providing this expertise which can supplement more traditional, extensive longitudinal management approaches.


In this review, we discuss the recent in-roads made into defining the molecular mechanisms and biochemical networks engaged by LEM and many other integral inner nuclear membrane proteins to preserve the nuclear barrier.The eukaryotic genome is enclosed in a nuclear envelope that protects it from potentially damaging cellular activities and physically segregates transcription and translation.Transport across the NE is highly regulated and occurs primarily  The eukaryotic genome is enclosed in a double membraned nucleus which serves to compartmentalize the genetic material and segregates processes like DNA replication and transcription processes from the cytosol. The nuclear envelope is continuous with the endoplasmic reticulum and is composed of two membranes, inner (INM) and outer nuclear membranes (ONM), separated by a lumen . TranspoGiven the complexity and size of the NPC, cellular machinery must ensure that individual nups are properly assembled to avoid any loss of function and, potentially, nuclear envelope integrity per se. As of now, four classes of proteins have been implicated in NPC assembly; 1) LEM (Lap2-emerin-Man1) and ESCRT  proteins, 2) lipid metabolism-related proteins, 3) membrane bending proteins, and 4) the scaffold nups .Saccharomyces cerevisiae and Schizosaccharomyces pombe, and this aids in sensing the NPC scaffold assembly state  and Heh2(Man1/LEMD3) in conjunction with the ESCRT machinery (The conserved Lap2-emerin-Man1 (LEM) family of integral INM proteins is required for tethering telomeres to the nuclear envelope and influences the integrity and structure of the nuclear envelope . In yeasachinery . Exposurachinery .S. cerevisiae, Heh2-GFP associates with the NPCs, as revealed by the fluorescence microscopy and immunoaffinity assays. However, in the absence of a properly formed scaffold in mutants for the outer ring complex nups , Heh2 fails to associate with those defective NPC. It is known that Heh2 binds to the NPC components via a winged helix domain located in its C-terminal nuclear domain /SpHeh1(Lem2) and their paralogue’s ScHeh2/SpHeh2(Man1)  . Both Sch2(Man1)  The evol of NPCs .APQ12 causes abnormal nuclear envelope morphology and defects in the assembly and distribution of NPCs as well as mRNA export  residing in the outer membrane and lead to a dynein-dependent SPB migration into the bud and NE breakage at the point of SPB attachment  and its reformation in an open form of mitotic cell division. This process begins with NPC permeabilization and NE invagination in the proximity of the centrosome followed by lamin network dissolution and NE-chromatin contact detachment along with the pulling forces of mitotic spindle entation  in the coformans , Ustilagiolaceae  and Ustio maydis , which utachment  rather tnogaster .To appreciate the complexity of NE rearrangements, we shall summarize the dynamics of metazoan NE remodeling during cell cycle and beyond to the best of our current knowledge in the following section.At the onset of metazoan mitosis, NE breaks down during prophase. By the end of anaphase, the fragmented NE is reassembled around decondensing chromatin to establish two individual daughter nuclear compartments. Broadly, the four major factors responsible for mitotic NEBD are: 1) Phosphorylation mediated loss in protein-protein interaction, 2) Spindle microtubule generated forces acting over the envelope, 3) Endoplasmic reticulum (ER) remodeling events and, 4) Chromatin-NE tethering perturbations. All of these operate in an overlapping fashion to orchestrate the mitotic NE remodeling  Figure .cdc2/CyclinB, PKC, and PKA, are also known to regulate lamin B1 polymerization to assist this NEBD  mediated disruption of anchoring proteins. Earlier, it was believed that chromatin detachment occurs passively during the breakdown process. On the contrary, recent studies in fission yeast have established an active function in regulating this event . The spetic exit .Multiple repair pathways operate in synchrony with the cell cycle to sustain NE integrity. The most efficient of which serves to re-establish NE between anaphase to telophase transition stage. Nevertheless, spontaneous rupture events are often documented during interphase like the defects mentioned in As mentioned above, the loss of NE integrity either while resealing the daughter nuclei as they exit mitosis or the rupture events that occur during interphase leads to a situation where the chromatin is left exposed to various genotoxic agents residing in the cytoplasm. Hence, upon NE rupture, multiple mutations are induced in the genome that might lead to carcinogenesis when the DNA repair machinery fails to re-establish its original nucleotide sequence. Further, NE rupture has been linked to micronuclei formation, protein mislocalization, chromatin rearrangement, and genomic instability events that give rise to cancer . MicronuDefects in NE other than NER that are documented as cancer predisposing factors like NE invagination and Nucleoplasmic reticulation (NR) can also occur in both dividing and resting cell nuclei . Since tIn addition to these NE deformations, other nuclear lamina and nuclear membrane proteins like Lamins, Nups, etc, contribute to oncogenesis mainly due to altered chromatin dynamics as a consequence of disturbed anchoring or epigenetic hallmarks of chromatin. For example, upon silencing lamin B1, the H3K27Me3 mark representing silent chromatin becomes more dispersed in the nuclear interior . Likewisvia chromatin-binding proteins (It is well established that the chromatin is often tethered to the nuclear periphery by means of nuclear lamina together with the nuclear envelope transmembrane proteins (NETs). This positioning is universal across cell types for certain facultative heterochromatin, but for others, it depends on the expression pattern of certain tethering proteins depending upon the state of differentiation . The verproteins . Proteinproteins , whereasproteins . Similarproteins  to estabproteins , Familiaproteins , Dunnigaproteins , Pelger-proteins  and Emerproteins , where Lproteins , FLH1 , ESCRT machinery, BAF (barrier-to-autointegration factor), and lamin proteins .via its curvature-sensitive membrane-binding domain but accumulates in the NE during NE reformation . In the absence of the NEC these viruses use NEBD as an alternative egress pathway to release the virions . SimilarDuring cell division, telomeric and centromeric-LINC interactions control NE disassembly by creating localized concentrations of factors that confer NE disassembly. This might be an evolutionary advantage to coordinate the preparedness of chromosomes and the onset of NEBD . Yet, th2+ ion concentration has also been shown to drive NEBD (2+ ion release is necessary for NEBD in mouse embryos (2+ ions in mammalian NEBD is still debated even though increased intracellular Ca2+ ion levels accelerate NEBD. So far, all these observations have only opened up more questions that remain open for future investigation.Although multiple tethering components involved in NEBD have been identified thus far, this list remains vastly incomplete today. Likewise, an increase in intracellular Caive NEBD , but con embryos . Hence, in vitro fertilization, and protoplast fusion-like scenarios while blocking this fusion might be of value in the context of viral replication in the future days to come.Future research in the field will improve our fundamental understanding of nuclear envelope biology that has broad implications for human health with probable outcomes in the form of new components and mechanistic insight into nuclear envelope repair pathways. Thus providing us with a better understanding of pathologies associated with immune decline, neurodegenerative diseases, viral infections, ageing, and laminopathies. Apart from cell biology, a better understanding of nuclear biology has translational potential in the form of designing new technologies for artificial membrane fusion required for targeted drug delivery systems,