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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.

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

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 (

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

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.

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.

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.

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.

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.

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.

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.

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.