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Molecular interactions between viral DNA and viral-encoded protein are a prerequisite for successful herpesvirus replication and production of new infectious virions. Here, we examined how the essential Kaposi’s sarcoma-associated herpesvirus (KSHV) protein, RTA, binds to viral DNA using transmission electron microscopy (TEM). Previous studies using gel-based approaches to characterize RTA binding are important for studying the predominant form(s) of RTA within a population and identifying the DNA sequences that RTA binds with high affinity. However, using TEM we were able to examine individual protein-DNA complexes and capture the various oligomeric states of RTA when bound to DNA. Hundreds of images of individual DNA and protein molecules were collected and then quantified to map the DNA binding positions of RTA bound to the two KSHV lytic origins of replication encoded within the KSHV genome. The relative size of RTA or RTA bound to DNA were then compared to protein standards to determine whether RTA complexed with DNA was monomeric, dimeric, or formed larger oligomeric structures. We successfully analyzed a highly heterogenous dataset and identified new binding sites for RTA. This provides direct evidence that RTA forms dimers and high order multimers when bound to KSHV origin of replication DNA sequences. This work expands our understanding of RTA binding, and demonstrates the importance of employing methodologies that can characterize highly heterogenic populations of proteins.ImportanceKaposi’s sarcoma-associated herpesvirus (KSHV) is a human herpesvirus associated with several human cancers, typically in patients with compromised immune systems. Herpesviruses establish lifelong infections in hosts in part due to the two phases of infection: the dormant and active phases. Effective antiviral treatments to prevent the production of new viruses are needed to treat KSHV. A detailed microscopy-based investigation of the molecular interactions between viral protein and viral DNA revealed how protein-protein interactions play a role in DNA binding specificity. This analysis will lead to a more in depth understanding of KSHV DNA replication and serve as the basis for anti-viral therapies that disrupt and prevent the protein-DNA interactions, thereby decreasing spread to new hosts. more...

Extracellular vesicles (EVs), or exosomes, play a pivotal role in tumor growth and metastasis, such as in the case of Kaposi Sarcoma. By loading tumor-derived EVs with chemotherapeutic drugs, we noted that their pro-tumor/pro-angiogenic phenotype was converted into an anti-tumor phenotype more...

We study the dynamics of threads of monodisperse droplets, including droplet chains and multi-chains, in which the droplets are interconnected by capillary bridges of another immiscible liquid phase. This system represents wet soft-granular matter - a class of granular materials in which the grains are soft and wetted by thin fluid films-with other examples including wet granular hydrogels or foams. In contrast to wet granular matter with rigid grains ( more...

Notre compréhension fondamentale de la biologie cellulaire s’est accélérée grâce à des décennies de recherche sur les divers mécanismes d’adaptation des virus à ADN pour survivre en exploitant leurs cellules hôtes. Les virus à ADN présentent une immense diversité, depuis les types de cellules qu’ils infectent jusqu’à la taille et la forme des virions. more...

PURPOSE This study aimed to determine the relationship between age of first exposure (AFE) to repetitive head impacts through contact/collision sports and patient-reported outcomes in community rugby players. METHODS We recruited community rugby players older than 18 yr with at least 1 yr of contact rugby participation to complete an online survey. Participants completed the Brief Symptom Inventory-18 (BSI-18), Short-Form Health Survey 12 (SF-12), and Satisfaction with Life Scale (SWLS) via Qualtrics. We used generalized linear models to examine the association between AFE (continuous) and patient-reported outcomes by sex, while controlling for cumulative years contact/collision sport history, age, and concussion history (yes/no). In addition, we used Mann-Whitney U tests to compare patient-reported outcomes between AFE more...

Background and aim Adenocarcinoma of the esophagus is still burdened by low 5-year survival. A complete pathologic response after neoadjuvant treatment is associated with improved long-term survival, however this result is rarely achieved. The aim of this study was to evaluate the impact of tumor regression grade (TRG), according to Mandard, on long-term survival. The primary endpoint was to determine whether the survival benefit associated with a complete tumor regression could be achieved also with a subtotal tumor response (TRG2). Secondly, we analyzed how TRG predicts survival in chemotherapy (CT) or chemoradiotherapy (CRT) treated patients and the relationship with nodal status (ypN). Methods We evaluated all consecutive patients who underwent esophagectomy for cancer at Padova University Hospital and Mainz University Hospital. Inclusion criteria were histologically confirmed ADK involving the thoracic esophagus or Siewert I and II cancers, treated by neoadjuvant CT or CRT and surgery. Patients with Siewert III or cervical cancer and those who underwent upfront surgery were excluded from the study. Kaplan-Maier analysis was used to estimate overall survival (OS) according to the TRG and ypN. Results Overall, 270 patients were recruited in the study period. The median OS was 50 months (range 1–134). TRG 1 and 2 were associated with improved survival (P Conclusion Tumor regression after neoadjuvant treatment is significantly associated with long term survival irrespective of the nodal status. In the overall study population and among CRT treated patients, the long-term survival was comparable between TRG 1 and TRG 2 patients. This finding allowed us to support the criterion of a Major Pathologic Response, which includes both TRG 1 and 2 ypN0 patients, potentially creating a homogeneous and larger subgroup of subjects to be evaluated in future clinical trials and studies. more...

Background and aim Laparoscopic Heller-Dor (LHD) is an effective treatment for esophageal achalasia, with good long-term results in more than 85–90% of patients. It remains controversial whether LHD is also indicated in the end-stage disease when the esophagus has a sigmoid shape. The aim of this study was to evaluate the outcome of LHD in patients with end-stage achalasia, as compared to patients with megaesophagus but without a sigmoid shape. Methods From 1992 to 2020, patients with a radiological diagnosis of sigmoid-shaped esophagus (radiological stage IV achalasia: group SE) and patients with straight, larger than 6 cm in diameter esophagus (radiological stage III achalasia: group NSE), both treated with LHD, were compared. Treatment failure was defined as the persistence or reoccurrence of an Eckardt score > 3, or the need for retreatment. Results During the study period, 164 such patients were observed: 73 patients in the SE group and 91 in the NSE group. The patients’ demographic and clinical parameters are summarised in Table 1. The median follow-up was 67 months (IQR: 24–106). No intra or postoperative mortality was recorded. There were 7 mucosal lesions: 2 in the SE group and 5 in the NSE group (P = 0.45). A positive outcome was obtained in 73% of patients in the SE group, as compared to 90% in the NSE group (P = 0.007, Table 2). All the patients whose surgery failed subsequently underwent one or more endoscopic pneumatic dilations. The overall success rate of the combination of LHD and endoscopic dilations was 83.6% in the SE group and all patients in the NSE group. The persisting failures had additional treatments: 4 patients a redo-myotomy, one each a botox injection, POEM and only one patient had an esophagectomy; the remaining 5 patients refused additional treatments. Postoperative 24-hour pH-monitoring was abnormal in 16.6% in SE group and 8.3% in NSE group (P = 0.33). Conclusion In end-stage achalasia patients, LHD may still achieve good results by relieving symptoms in more than 70% of patients. Therefore, LHD may still be the first surgical option to be offered to these patients before esophagectomy. more...

Background and aim Esophageal perforation (EP) is characterized by high morbidity and mortality. In recent years, non-operative management has proved to be successful in selected patients. However, there is a lack of data supporting the superiority of one approach over the others and there is no consensus on the treatment algorithm. The Pittsburgh Severity Score (PSS) is a scoring system based on clinical factors at the time of EP presentation, intended to guide treatment. The aim of the study is to verify PSS usefulness in stratifying EP severity and in guiding clinical decisions. Methods All patients referred to our Unit for EP between January 2005 and January 2020 were enrolled. Patients were stratified according to their PSS into 3 groups (PSS ≤ 2; 3–5 and > 5): the postoperative outcomes were compared. The predictive value of the PSS was evaluated by simple linear and logistic regression for the following outcomes: need for surgery, complications, in-hospital mortality, ICU and hospital stay, time to refeeding and need for reintervention. Results Seventy-three patients were referred for EP (M/F: 46/27). Perforations were more frequently iatrogenic (41.1%) or spontaneous (37.0%). The median PSS was 4 (IQR 2–6). Surgery was required in 60.3% of cases. The median PSS was significantly lower in the conservative group (P Conclusion PSS is useful in stratifying patients in risk groups with different morbidity and mortality. It is also useful in guiding the therapeutic conduct, selecting patients for nonoperative management. Prospective studies are needed to confirm the role of the PSS in the treatment of esophageal perforation. more...

Background and aim The current standard of care in treating locally-advanced esophageal cancer has been established after the publication of the landmark CROSS trial and several others, which demonstrated an overall survival benefit from preoperative chemoradiation prior to esophagectomy in this setting. However, elderly patients have been generally excluded from randomized study protocols, so literature regarding the choice of the best treatment approach for resectable esophageal cancer in patients older than 75 years old is limited. The aim of this study was to characterize outcomes associated with neoadjuvant chemoradiation prior to esophagectomy, compared to esophagectomy alone, in older patients with esophageal cancer. Methods We conducted a retrospective study in patients ≥75 years with locally-advanced esophageal cancer (both adenocarcinoma -EAC and squamous cell carcinoma -SCC) undergoing esophagectomy ± neoadjuvant chemoradiation at our Center between 2000 and 2020. Results A total of 141 patients were included in the study. Patients were subcategorized into two groups: patients undergoing upfront surgery (SURG, n = 97) and patients undergoing neoadjuvant therapy before surgery (NAT, n = 44). Patients in the NAT group experienced a significantly higher postoperative morbidity when compared to the SURG group. Overall survival (21 months and 25 months, p = 0.8883) and cancer-related survival (20 and 25 months, p 0.8052) were not statistically different between the SURG and the NAT group. When looking to the histology, OS for EAC and for SCC did not significantly differ between the 2 groups. We subcategorized patients according to their final pathological stage into patients with pTNM stage I and II, and patients with stage III and IV. Among patients with EACs stage III-IV, the NAT group experienced a significant benefit in OS when compared to the SURG group. Univariate analysis for risk factors for postoperative morbidity showed a significant association between NAT and postoperative complications (p 0,0336). Multivariate analysis confirmed neoadjuvant therapy as an independent prognostic factor for morbidity (P 0,0226). Conclusion In our study, patients undergoing NAT before surgery experienced a higher postoperative morbidity when compared to patients undergoing upfront esophagectomy. Further studies are needed to identify which older patients are most suitable for neoadjuvant therapy before esophagectomy. more...

Background and aim Esophago-gastric junction outflow obstruction (EGJOO) is a relatively new clinical entity revealed by high-resolution manometry (HRM) that can have two different etiologies, i.e. idiopathic or secondary (esophageal strictures, eosinophilic esophagitis, giant hiatal hernia, prior fundoplication or bariatric surgery, malignancy). Treatment of EGJOO currently mirrors that of achalasia, but this is based only on few studies with small case series. The aim of this study was to assess the outcome of laparoscopic Heller-Dor (LHD) in patients with EGJOO, as compared with patients with esophageal achalasia. Methods Between 2016–2019, all patients with a diagnosis of idiopathic EGJOO and referred to our Centre for dysphagia or food regurgitation were prospectively collected. Naïve EGJOO patients were first given medical treatment with Ca2+ channel blockers for 6 months. Then patients were ‘restaged’: patients without clinical improvement and/or whose integrated relaxation pressure (IRP) had not decreased were scheduled for LHD. EGJOO patients who had already been treated elsewhere with botox injections, pneumatic dilations or Ca2+ channel blockers were scheduled upfront for LHD. Patients with EGJOO who underwent LHD were compared with patients with radiological stage I achalasia. The achalasia group was further analyzed by subgrouping the patients based on the manometric-pattern. Treatment failure was defined as the persistence or reoccurrence of a Eckardt score > 3 or the need for retreatment. Results During the study period, 150 patients were enrolled: 25 EGJOO patients and 125 radiological stage I achalasia (25 pattern I, 74 pattern II, and 26 pattern III). The median follow-up was 24 months (IQR: 34–16). Treatment was successful in 96% of patients in the EGJOO group and in 96% of achalasia patients with pattern I, 98.7% in those with pattern II, and 96.2% of those with pattern III (P = 0.50). Six-month post-operative HRM showed a reduction in the LES resting pressure and IRP for all patients in all 4 groups (P Conclusion This is the first comparative study based on prospective data collection to assess the outcome of LHD in patients with EGJOO. LHD emerged as an effective treatment for EGJOO, with an excellent success rate, comparable with the procedure’s efficacy in treating early-stage achalasia. more...

Background: Clinical and epidemiological data showed that African American (AA) women suffer higher incidence and mortality rates due to triple-negative breast cancer (TNBC) compared to European American (EA) women. These disparate disease outcomes are attributed to multiple factors including socioeconomic status, access to health care, and earlier disease onset. However, a growing body of evidence also shows the existence of different gene expression signatures and tumor microenvironments between these patient racial groups. We and others identified CRYβB2 as a cancer racial disparity gene that has higher expression in AA patients with breast, colon, and prostate cancers. We previously demonstrated, via in vitro and in vivo studies, that CRYβB2 promotes proliferation, metastasis, and IL-6 signaling. The objective of the current research is to identify the cellular and molecular mechanisms downstream of CRYβB2 leading to its tumor-promoting phenotypes. Methods: CRISPR/Cas9 and lentiviral knock-in gene editing tools were used to create TNBC cell lines with CRYβB2 knockout and overexpression respectively. The functional consequences of CRYβB2 were assessed in vitro using colony formation, tumor spheroid invasion, and collagenase activity assays. Additionally, tumor spheroids were grown and RNA sequencing (RNA-seq) was performed to assess the effect of CRYβB2 on gene expression. Results: Herein, functional assays showed that CRYβB2 promotes proliferation and invasion in vitro in both AA and EA cell lines. RNA-seq and Gene Ontology enrichment analyses revealed that CRYβB2 alters key pathways such as cell adhesion and extracellular matrix (ECM) organization. RT-qPCR validation also showed that CRYβB2 inhibits the expression of a specific class of adhesion molecules known as protocadherins (PCDHs). Downregulation of PCDHs has been shown to enhance tumor progression and is a predictive marker for low survival in many cancer types. Indeed, data mining of publicly available data shows that three of the identified PCDHs (PCDHGB5, PCDHGB6 and PCDHGB7) have significantly lower expression levels in breast cancer compared to normal tissue and their low expression leads to low overall survival. Furthermore, RNA-seq data showed that CRYβB2 promotes the expression of multiple matrix metalloproteinase (MMP) genes. This was functionally validated using collagenase activity assay where CRYβB2 enhanced collagen degradation in vitro. Conclusions: Our findings provide a mechanistic explanation for the role of CRYβB2 in driving disparate clinical outcome among AA and EA TNBC patients. We demonstrate a paradigm where high CRYβB2 expression promotes tumor progression by suppressing cell adhesion molecules and increasing ECM degradation to enhance tumor growth, invasion, and metastasis. Disclaimer: This work was prepared while Jodie Fleming was employed at NCCU. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. Citation Format: Amr A. Waly, London Harper, Jodie M. Fleming, Lindsey M. Costantini. The racial disparity-linked gene CRYβB2 alters cell adhesion and tumor microenvironment to promote proliferation and invasion in triple-negative breast cancer [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PR006. more...

Introduction Military members’ knowledge of concussion signs and symptoms may be critical to appropriate concussion identification and health-seeking behavior, particularly for those in leadership roles. The current study aimed to characterize concussion knowledge and attitudes among future military officers undergoing U.S.-based Reserve Officers’ Training Corps (ROTC) training. Materials and Methods Army and Air Force ROTC cadets at 2 large, public universities were utilized for a survey-based observational study. The study was approved by the institutional review board at both university research sites. Cadets completed a modified Rosenbaum Concussion Knowledge and Attitude Survey to obtain cadets’ Concussion Knowledge Index and Concussion Attitude Index, where higher scores are preferable. Cadets’ concussion knowledge and attitudes were characterized via descriptive statistics. Results Cadets (n = 110) had a mean Concussion Knowledge Index of 18.8 ± 3.2 (range = 9–23, out of 25). Potentially detrimental misconceptions included: belief that typically concussion symptoms no longer persist after 10 days (79.1%) and brain imaging shows visible physical damage following concussion (74.5%). Mean Concussion Attitude Index was 60.6 ± 7.4 (range = 46–75, out of 75). In general, cadets reported higher agreement with safe concussion behavior than what they believe peers would report. Conclusions Cadets were found to have a high concussion knowledge, yet common misconceptions remained. Cadets consistently reported safe choices but were less sure that peers felt similarly; future investigations should evaluate ROTC concussion social norms and education should note peers’ beliefs supporting safe concussion attitudes. more...

This study aimed to determine the relationship between age of first exposure (AFE) to repetitive head impacts through contact/collision sports and patient-reported outcomes in community rugby players.We recruited community rugby players older than 18 yr with at least 1 yr of contact rugby participation to complete an online survey. Participants completed the Brief Symptom Inventory-18 (BSI-18), Short-Form Health Survey 12 (SF-12), and Satisfaction with Life Scale (SWLS) via Qualtrics. We used generalized linear models to examine the association between AFE (continuous) and patient-reported outcomes by sex, while controlling for cumulative years contact/collision sport history, age, and concussion history (yes/no). In addition, we used Mann-Whitney U tests to compare patient-reported outcomes between AFE12 and AFE ≥12.A total of 1037 rugby players (31.6 ± 11.3 yr (range, 18-74 yr), 59.1% men) participated in this study. Whether analyzed continuously or dichotomously at age 12 yr, younger AFE was not associated with worse patient-reported outcomes for either men or women. Positive concussion history was a significant predictor of worse BSI-18 subscores, SF-12 subscores, and SWLS in women and worse BSI-18 subscores in men. Cumulative contact/collision sport history was a significant predictor of better BSI-18 Depression and SF-12 (Mental Component Summary) subscores in men only. In men and women, older age was a significant predictor of better BSI-18 Depression, Anxiety, and GSI subscores; better SWLS (in men only); and better SF-12 Mental Component Summary, but worse SF-12 (Physical Component Summary).Younger AFE to contact/collision sport is not associated with worse patient-reported outcomes in early adult rugby players. Concussion history was predictive of worse patient-reported outcomes. more...

To determine (1) the association between lifetime diagnosed concussion and lower extremity musculoskeletal injury (LE-MSI) among community rugby union players and (2) the sex specific risk of LE-MSI given concussion history among males and females.Retrospective survey.1037 (59.0% male, (612/1037), age: 31.6 ± 11.3 years) rugby players (10.1 ± 8.1 years played) completed an online survey to ascertain injury history. A chi-squared test of association was performed between concussion and LE-MSI; significant outcomes were followed-up with an odds ratio. A binary logistic regression with any LE-MSI (yes/no) as the outcome and concussion (yes/no) and sex (male/female) as predictors was performed to determine if there was a sex by concussion interaction.There was an overall significant association between concussion and any LE-MSI(χ(1) = 13.055, p 0.001, OR = 2.30 [95%CI: 1.45, 3.65]). Both male (OR = 2.21) and females (OR = 2.49) had significant associations for concussion and LE-MSI, but there were no differences between sex for risk of LE-MSI (RCommunity rugby players with a history of concussion are2× more likely to also experience an LE-MSI than those without a history of concussion. There were no differences in the odds of LE-MSI between males and females with a history of diagnosed concussion. In line with current World Rugby injury prevention programs, future research should aim to reduce LE-MSI incidence to maximize player safety and wellness through targeted injury prevention and teams should utilize a conservative return to play protocols following concussion. more...

IntroductionConcussions have been associated with an increased risk of lower-extremity musculoskeletal injury (LE-MSI) in athletes and US Army soldiers, creating an added economic, physical and social burden. Yet, there is a paucity of evidence on this relationship among Reserve Officers’ Training Corps (ROTC) cadets, a group which engages in activities with high-injury risk and will subsequently commission as active duty officers. This study aimed to examine the association between concussions and LE-MSI in ROTC cadets.Methods125 (83 were male) Army and Air Force ROTC cadets (19.8±2.0 years) from two large state universities’ Army and Air Force ROTC programmes participated in this study. Cadets completed a reliable injury history questionnaire to ascertain the following variables of interest: (1) any concussion history, (2) reported concussions, (3) undiagnosed concussions, and (4) potentially unrecognised concussion history and LE-MSI history (eg, ankle sprain, knee sprain or muscle strain). Data were analysed using a χ2test for association and binary logistic regression to determine ORs.ResultsCadets with any concussion history (n=42) had a significantly (p=0.035) higher association with LE-MSI (OR 2.47, 95% CI 1.05 to 5.83) than those without. Cadets who had a reported concussion (n=33) had a significantly (p=0.026) higher association with LE-MSI (OR 2.95, 95% CI 1.11 to 7.84) compared to cadets without.ConclusionsROTC cadets with a history of diagnosed concussion were more likely to have suffered an LE-MSI than cadets without a concussion history. ROTC cadre should be aware of this relationship and incorporate injury prevention protocols. more...

In this article, I explore the performative nature of ghost bike memorials across the United States. Ghost bikes, flat-white painted immobile bicycles, are installed roadside to mark the location o... more...

To provide a summary of the evidence on the comparative effectiveness of two surgical treatment strategies, sentinel node biopsy (SNB) and elective neck dissection (END), in patients with T1-T2 oral cancer and clinically negative (cN0) neck, in terms of overall survival (OS), disease-free survival (DFS) and neck recurrence rates (NRRs).A systematic review was performed by including studies published up to April 2019. Meta-analysis was performed to compare NRRs between SNB and END. A narrative summary of the results was generated for OS, DFS and morbidity outcomes. The certainty of evidence was assessed according to the GRADE methodology.No randomized studies were retrieved. Five observational studies were included in the comparative effectiveness analysis and four observational studies were included in the comparative morbidity analysis. The pooled risk ratio showed no differences in NRRs between SNB and END (10.5% vs 11.6%; pooled RR 1.09; 95% CI 0.67-1.76). No differences in OS or DFS between the two treatments were found. SNB appears to be associated with a lower rate of postoperative complications and lower shoulder dysfunction than END. Conversely, the results of the quality of life (QoL) questionnaires are not sufficient to advocate a particular strategy.Our review highlights the lack of well conducted and randomized studies comparing SNB to END, leading to poor clinical evidence. Although our findings suggest no significant differences in OS, DFS and NRR between the two strategies, the certainty of our evidence is too low to make it useful for clinical decision making. more...

Background In the last decade of the past century, primary Laparoscopic Heller-Dor (LHD) for Achalasia progressively became the procedure of choice in the new millennium. The aim of this study was to assess the long-term outcome of LHD to treat Achalasia at a single high-volume institution during the past 25 years. Methods 1000 patients underwent LHD from 1992-2017 by 6 staff surgeons alternatively. Patients who had already been treated with surgical or endoscopic myotomy were ruled out. Symptoms were collected and scored using a detailed questionnaire; barium-swallow, endoscopy, manometry were performed, before and after surgery while, 24-hour pH monitoring were performed 6 months after surgery. Results LHD was the primary treatment for 1000 patients (M:F=536:464); the median age was 46 (IQR 36-54), 183 (18.3%) had a history of endoscopic treatments (pneumatic dilation or botox injections, or both). The surgical procedure was completed laparoscopically in all but 7 patients (0.7%) and there was one perioperative death for heart attack. There were 25 perforations (2.5%): 22 were recognized and repaired during the operation, 3 were detected by postoperative contrast swallow. The outcome was positive in 902 patients (90.2%). In patients who had a previous treatment the failures were 25/183 (13.7%) while in the primary treatment group the failures were 73/817 (8.9%) (p=0.055). All the 98 patients whose LHD failed subsequently underwent one or more endoscopic pneumatic dilations, which ameliorated their recurrent symptoms in all but 11 patients (10 of whom required reoperation). The overall success rate of the combination of LHD and endoscopic dilations (where necessary) was 98.4%. At univariate analysis, manometric pattern (p=0.001), sigmoid megaesophagus (p=0.003) and a chest pain score (p=0.002) were the only factors predictive of a positive final results. At multivariate analysis, these three factors were independently associated to good outcome. Postoperative 24-hour pH-monitoring was abnormal in 50/590 patients (8.5%) Two patients developed an esophageal cancer during the follow-up time. Conclusions In a university tertiary referral center LHD can durably relieve achalasia symptoms. Preoperative manometric pattern, a presence of a sigmoid esophagus and the chest pain score represent the strongest predictor of outcome. more...

This data article is related to the research article entitled “Silver nanoparticles alter epithelial basement membrane integrity, cell adhesion molecule expression and TGF-beta secretion”, available in the journal Nanomedicine: Nanotechnology, Biology, and Medicine [1]. This Data in Brief consists of data that describe changes in the expression of basement membrane (BM)-associated genes and proteins in three non-transformed epithelial cell lines following acute (6 h) and chronic (24 h plus 7-day chase) exposure to silver nanoparticles (AgNPs). Human BEAS2B (lung), MCF10AI (breast), and CCD-18Co (colon) cultured epithelia were analyzed for protein expression by LC-MS/MS and for gene expression by pathway-focused QRT-PCR arrays of 168 focal adhesion, integrin, and extracellular matrix (ECM) genes known to be localized to the plasma membrane, the BM/ECM, or secreted into the extracellular space. Ingenuity pathway analysis (IPA) of combined gene and protein expression datasets was then used to predict canonical pathways affected by AgNP exposure. Keywords: Silver nanoparticles, Epithelial cells, Basement membrane, Extracellular matrix, TGF-Beta, Intracellular signalling pathway analysis more...

ObjectiveTo determine the relationship between exposure to repetitive head impacts (RHI) through collision sports prior to the age of 12 and quality of life measures in community rugby players.BackgroundIt is suggested that RHI incurred before age 12 may be associated with later life neurologic impairments. However, research on age of first exposure (AFE) to collision sports and psychological outcomes has not be explored in rugby, a sport which participants often continue in community settings beyond college.Design/MethodsIndividuals over 18 years old who currently or previously played contact rugby completed an online questionnaire. To assess quality of life and psychological status, participants completed the Brief-Symptoms Inventory 18 (BSI-18), Short Form 12 (SF-12), and Satisfaction with Life Survey (SWLS). Participants were dichotomized into AFE to collision sports (12); AFE to rugby was not used since most participated in other collision sports prior to rugby. Data were not normally distributed; therefore, a Mann-Whitney U test was performed to compare outcomes between AFE groups.Results1,037 rugby players (31.6 + 11.3 years, 59.1% male) participated in this study. There were no significant differences between AFE 12 groups on all outcomes: BSI-18 Somatization (U = 97,286, p = 0.307), BSI-18 Depression (U = 100,267, p = 0.778), BSI-18 Anxiety (U = 98,851, p = 0.531), SF-12 Physical (U = 94,413, p = 0.241), SF-12 Mental (U = 96,517, p = 0.512), SWLS (U = 98,866, p = 0.537). Mean scores for all outcomes were: BSI-18 Somatization (2.33 + 2.99), BSI-18 Depression (4.20 + 4.91), BSI-18 Anxiety (3.32 + 3.75), SF-12 Physical (52.40 + 7.25), SF-12 Mental (46.20 + 11.45), SWLS (24.86 + 6.31).ConclusionsConsistent with recent cohort studies, there was no observed difference on three common measures of psychological well-being and quality of life in rugby players based upon AFE to collision sports. However, later life potential consequences of RHI in rugby players remains to be elucidated. more...

ObjectiveTo examine sex differences between concussion and lower-extremity musculoskeletal injury (LE-MSI) in community male and female rugby players.BackgroundThere is an ∼2x elevated risk of post-concussion subsequent MSI in high school through professional athletes. However, the effect of sex on risk is inconsistent and sparse, and rugby provides an ideal population as it’s the only collision sport with the same rules for both sexes.Design/Methods1,037 rugby players (31.6 + 11.3 years, 59.1% male), with at least one year of rugby playing experience, participated in this study, completing an online injury history questionnaire to ascertain concussion (yes/no) and LE-MSI (yes/no) history. A chi-squared test was performed to determine the association between concussion and any LE-MSI; significant findings were followed up with a post hoc odds ratio test. A binary logistic regression with any LE-MSI (yes/no) as the outcome and concussion (yes/no) and sex (male/female) as predictors was performed to determine if there was a sex by concussion interaction.ResultsThere was a significant association between concussion and any LE-MSI for all groups (Overall: ?(1) =13.06, p < 0.001, OR = 2.30 [95% CI: 1.45–3.65]; Males: ?(1) =7.43 p = 0.006, OR = 2.21 [95% CI: 1.24–3.96]; and Females: ?(1) = 5.78, p = 0.016, OR = 2.48 [95% CI: 1.16–5.31]). However, there were no differences for risk of LE-MSI between males and females (p = 0.99, R2 = 0.024).ConclusionsBoth male and female community rugby players had a 2x greater risk of LE-MSI, given a history of concussion compared to those without a history of concussion, which aligns with previous studies focused on collegiate athletes. However, there was no difference in risk of LE-MSI between sexes, contrary to smaller, but more controlled studies. Future research should investigate the potential physiological mechanisms for increased risk of LE-MSI. more...

International audience; Single crystals of transition metal carbides from group IV, TiC 1-x and ZrC 1-x , have been irradiated at room temperature (RT) with 1.2 MeV gold ions for various fluences in the range 2 × 10 14 –3 × 10 16 ions/cm 2 and polycrystals for fluences between 2 × 10 13 and 4 × 10 15 ions/cm 2 . The irradiated samples were characterized by micro Raman spectroscopy. For this purpose, the evolution of main parameters of Raman spectra for both types of samples (band positions, shifts, and intensities, and optical over acoustical band area ratios) versus ion fluence are reported, and discussed. All acoustic and optical bands increase and broaden with fluence. Yet no amorphization is achieved under the highest fluence for both carbides. From the lowest fluence up to 10 15 ions/cm 2 , point defects like interstitials and vacancies are created. The increasing number of carbon vacancies involves a local variation in stoichiometry accompanied by carbon release. For fluence above 10 15 ions/cm 2 , extended defects like dislocation loops appear, as seen by TEM observations. more...

Isolation of extracellular vesicles (EVs) from cell culture supernatant or plasma can be accomplished in a variety of ways. Common measures to quantify relative success are: concentration of the EVs, purity from non-EVs associated protein, size homogeneity and functionality of the final product. Here, we present an industrial-scale workflow for isolating highly pure and functional EVs using cross-flow based filtration coupled with high-molecular weight Capto Core size exclusion. Through this combination, EVs loss is kept to a minimum. It outperforms other isolation procedures based on a number of biochemical and biophysical assays. Moreover, EVs isolated through this method can be further concentrated down or directly immunopurified to obtain discreet populations of EVs. From our results, we propose that cross-flow/Capto Core isolation is a robust method of purifying highly concentrated, homogenous, and functionally active EVs from industrial-scale input volumes with few contaminants relative to other methods. more...

Debilitating heart conditions, notably dilated and hypertrophic cardiomyopathies (CM), are associated with point mutations in metavinculin, a larger isoform of the essential cytoskeletal protein vinculin. Metavinculin is co-expressed with vinculin at sub-stoichiometric ratios in cardiac tissues. Cardiomyopathy mutations in the metavinculin tail domain (MVt) occur within the extra 68-residue insert that differentiates it from the vinculin tail domain (Vt). Vt binds actin filaments (F-actin) and promotes vinculin dimerization to bundle F-actin into thick fibers. While MVt binds to F-actin in a similar manner to Vt, MVt is incapable of F-actin bundling and inhibits Vt-mediated F-actin bundling. We performed F-actin co-sedimentation and negative-stain EM experiments to dissect the coordinated roles of metavinculin and vinculin in actin fiber assembly and the effects of three known metavinculin CM mutations. These CM mutants were found to weakly induce the formation of disordered F-actin assemblies. Notably, they fail to inhibit Vt mediated F-actin bundling, and instead promote formation of large assemblies embedded with linear bundles. Computational models of MVt bound to F-actin suggest that MVt undergoes a conformational change licensing the formation of a protruding sub-domain incorporating the insert, which sterically prevents dimerization and bundling of F-actin by Vt. Sub-domain formation is destabilized by CM mutations, disrupting this inhibitory mechanism. These findings provide new mechanistic insights into the ability of metavinculin to tune actin organization by vinculin and suggest that dysregulation of this process by CM mutants could underlie their malfunction in disease. more...

Extracellular vesicles (EVs) or exosomes have been implicated in the pathophysiology of infections and cancer. The negative regulatory factor (Nef) encoded by simian immunodeficiency virus (SIV) and human immunodeficiency virus (HIV) plays a critical role in the progression to AIDS and impairs endosomal trafficking. Whether HIV-1 Nef can be loaded into EVs has been the subject of controversy, and nothing is known about the connection between SIV Nef and EVs. We find that both SIV and HIV-1 Nef proteins are present in affinity-purified EVs derived from cultured cells, as well as in EVs from SIV-infected macaques. Nef-positive EVs were functional, i.e., capable of membrane fusion and depositing their content into recipient cells. The EVs were able to transfer Nef into recipient cells. This suggests that Nef readily enters the exosome biogenesis pathway, whereas HIV virions are assembled at the plasma membrane. It suggests a novel mechanism by which lentiviruses can influence uninfected and uninfectable, i.e., CD4-negative, cells., IMPORTANCE Extracellular vesicles (EVs) transfer biologically active materials from one cell to another, either within the adjacent microenvironment or further removed. EVs also package viral RNAs, microRNAs, and proteins, which contributes to the pathophysiology of infection. In this report, we show that both human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) incorporate the virus-encoded Nef protein into EVs, including EVs circulating in the blood of SIV-infected macaques and that this presents a novel mechanism of Nef transfer to naive and even otherwise non-infectable cells. Nef is dispensable for viral replication but essential for AIDS progression in vivo. Demonstrating that Nef incorporation into EVs is conserved across species implicates EVs as novel mediators of the pathophysiology of HIV. It could help explain the biological effects that HIV has on CD4-negative cells and EVs could become biomarkers of disease progression. more...