The dopaminergic system is implicated in the pathophysiology of migraine. However, the underlying mechanisms remain unclear. We explored the effects and mechanisms of dopaminergic system modulation in the in-vivo and in-vitro rat models of migraine. Dopaminergic agonist apomorphine, D2 receptor antagonists metoclopramide and haloperidol, and 5-HT3 receptor antagonist ondansetron alone and together were tested in nitroglycerin-induced migraine model, in vivo. Likewise, the combinations of drugs were also tested on basal CGRP release in-vitro hemiskull preparations. Mechanical allodynia was tested by von-Frey filaments. CGRP concentrations in trigeminovascular structures and in-vitro superfusates, and c-Fos levels in brainstem were determined by ELISA. Meningeal-mast cells were evaluated with toluidine-blue staining. Apomorphine further enhanced nitroglycerin-induced mechanical allodynia, brainstem c-fos expression, trigeminal ganglion and brainstem CGRP concentrations, and meningeal mast cell degranulation, in vivo. Haloperidol completely antagonised all apomorphine-induced effects and also alleviated changes induced by nitroglycerin without apomorphine. Metoclopramide and ondansetron partially attenuated apomorphine- or nitroglycerin-induced effects. A combination of haloperidol and ondansetron decreased basal CGRP release, in-vitro, while the other administrations were ineffective. Apomorphine-mediated dopaminergic activation exacerbated nitroglycerin-stimulated migraine pain by further enchancing c-fos expression, CGRP release and mast cell degranulation in strategical structures associated with migraine pain. Metoclopramide partially attenuated the effects of apomorphine, most likely because it is also a 5-HT3 receptor antagonist. Haloperidol with pure D2 receptor antagonism feature appears to be more effective than metoclopramide in reducing migraine-related parameters in dopaminergic activation- and/or NTG-induced migrane like conditions.

Background: It is unknown whether Torque Teno virus (TTV) DNA load monitoring could anticipate the development of infectious events in hematological patients undergoing treatment with small molecular targeting agents. We characterized the kinetics of plasma TTV DNA in patients treated with ibrutinib or ruxolitinib and assessed whether TTV DNA load monitoring could predict the occurrence of Cytomegalovirus (CMV) DNAemia or the magnitude of CMV-specific T-cell responses. Methods: Multicenter, retrospective, observational study, recruiting 20 patients treated with ibrutinib and 21 with ruxolitinib. Plasma TTV and CMV DNA loads were quantified by real-time PCR at baseline and days +15, +30, +45, +60, +75, +90, +120, +150, and +180 after treatment inception. Enumeration of CMV-specific IFN-γ-producing CD8 + and CD4 + T cells in whole blood was performed by flow cytometry. Results: Median TTV DNA load in ibrutinib-treated patients increased significantly ( P=0.025) from baseline (median, 5.76 log 10 copies/ml) to day +120 (median, 7.83 log 10 copies/ml). A moderate inverse correlation (Rho=-0.46; P<0.001) was found between TTV DNA load and absolute lymphocyte count (ALC). In ruxolitinib-treated patients, TTV DNA load quantified at baseline was not significantly different from that measured after treatment inception ( P ≥0.12). TTV DNA loads were not predictive of the subsequent occurrence of CMV DNAemia in either patient group. No correlation was observed between TTV DNA loads and CMV-specific IFN-γ-producing CD8 + and CD4 + T cell counts in either patient group. Conclusion: The data did not support the hypothesis that TTV DNA load monitoring in hematological patients treated with ibrutinib or ruxolitinib could be useful to predict either the occurrence of CMV DNAemia or the level of CMV-specific reconstitution.

A document by Carlos F de Paule. Click on the document to view its contents.

A document by Yasmin Sheikhhassani. Click on the document to view its contents.

Liposomal drug products are playing an increasing role in the field of drug delivery. With this increased demand comes the need to increase the capabilities and capacity of manufacturing options. Continuous manufacturing techniques present a significant opportunity to address these needs for liposomal manufacturing processes. Liposomal formulations have unique considerations that impact translation from batch to continuous process designs. This article examines aspects of converting to a continuous design that were previously viewed as inconsequential in a batch process. The batch process involves the removal of ethanol through tangential flow filtration (TFF). Ethanol was found to reduce the permeability of the hollow fibers used for TFF. This effect was determined to have minimal impact on the overall batch process design but considerable influence on the design of continuous TFF such as inline diafiltration (ILDF). Using a pilot scale setup, ethanol was found to decrease permeability in an inverse manner to ethanol concentration. Further assessment found that dilution of the ethanol levels prior to diafiltration can significantly reduce the amount of ILDF stages needed and that a continuous design requires less buffer to the commensurate batch design.

A document by Bea Andersson. Click on the document to view its contents.

Cell-derived extracellular vesicles (EVs) are evolutionary-conserved secretory organelles that, based on their molecular composition, are important intercellular signaling regulators. At least three classes of circulating EVs are known based on mechanism of biogenesis: exosomes (sEVs/Exos), microparticles (lEVs/MPs) and shed midbody remnants (sMB-Rs). sEVs/Exos are of endosomal pathway origin, microparticles (lEVs/MPs) from plasma membrane blebbing, and shed midbody remnants (sMB-Rs) arise from symmetric cytokinetic abscission. Here, we isolate sEVs/Exos, lEVs/MPs and sMB-Rs secreted from human isogenic primary (SW480) and metastatic (SW620) colorectal cancer (CRC) cell lines in milligram quantities for label-free MS/MS-based proteomic profiling. Purified EVs revealed selective composition packaging of exosomal protein markers in SW480/SW620-sEVs/Exos, metabolic enzymes in SW480/SW620-lEVs/MPs, while centralspindlin complex proteins, nucleoproteins, splicing factors, RNA granule proteins, translation-initiation factors, and mitochondrial proteins selectively traffic to SW480/SW620-sMB-Rs. Collectively, we identify 39 human cancer-associated genes in EVs; 17 associated with SW480-EVs, 22 with SW620-EVs. We highlight oncogenic receptors/transporters selectively enriched in sEVs/Exos (EGFR/ FAS in SW480-Exos and MET, TGFBR2, ABCB1 in SW620-sEVs/Exos). Interestingly, MDK, STAT1, and TGM2 are selectively enriched in SW480-sMB-Rs, and ADAM15 to SW620-sMB-Rs. Our study reveals sEVs/Exos, lEVs/MPs and sMB-Rs have distinct protein signatures that open potential diagnostic avenues of distinct types of EVs for clinical utility.

Pemigatinib treatment for intrahepatic cholangiocarcinoma with FGFR2 fusion detected by a liquid comprehensive genomic profiling testShun Ishido¹*, Nobuharu Tamaki¹*, Kento Inada¹, Jun Itakura¹, Yuka Takahashi¹, Naoki Uchihara¹, Keito Suzuki¹, Yuki Tanaka¹, Haruka Miyamoto¹, Michiko Yamada¹, Hiroaki Matsumoto¹, Tsubasa Nobusawa¹, Taisei Keitoku¹, Kenta Takaura¹, Shohei Tanaka¹, Chiaki Maeyashiki¹, Yutaka Yasui¹, Kaoru Tsuchiya¹, Hiroyuki Nakanishi¹, Masayuki Kurosaki¹#, Namiki Izumi¹*These authors equally contributed to this case report.#This author (Masayuki Kurosaki) is corresponding author.Department of Gastroenterology and Hepatology, Musashino Red Cross Hospital, Tokyo, JapanGuarantor of this article: Masayuki Kurosaki

Objectives: We recently reported that deep learning (DL) using pelvic magnetic resonance imaging is useful for predicting the severity of urinary incontinence (UI) after robot-assisted radical prostatectomy (RARP). However, our results were limited because the prediction accuracy was approximately 70%. We aimed to develop a more accurate prediction system that can be used to inform patients on recovery from UI after RARP using a DL model based on intraoperative video images. Materials and Methods: This study included 101 patients with prostate cancer who underwent RARP. Three snapshots showing the pelvic cavity (before bladder neck incision, just after prostate removal, and after vesicourethral anastomosis) from intraoperative video records, as well as preoperative and intraoperative covariates, were assessed. We evaluated the DL models plus simple or ensemble machine learning, and their sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) were analyzed. Results: Sixty-four and 37 patients demonstrated ‘early continence’ and ‘late continence’, respectively, at the 3-month follow-up. The combination of DL and simple machine learning using intraoperative video snapshots with clinicopathological parameters had a notably high performance (AUC, 0.683 to 0.749) for predicting early recovery from post-prostatectomy UI. Notably, the combination of DL and ensemble artificial neural network using intraoperative video snapshots had the highest performance (AUC, 0.882; sensitivity, 92.2%; specificity, 78.4%; overall accuracy, 85.3%) for predicting early recovery from post-prostatectomy UI. In contrast, DL and ensemble ML with clinicopathological parameters (Method 4) achieved no additive effects (AUC, 0.690 to 0.747) compared with DL and simple ML with clinicopathological parameters. Internal validation was performed on additional 30 cases with similar results. Conclusions: Our results suggest that DL algorithms using intraoperative video images can be used to reliably inform patients regarding their recovery from UI after RARP. (287 words)

Introduction Randomised controlled trials show a reduction in acute kidney injury, renal impairment, and acute renal failure after initiation of a sodium glucose cotransporter-2 inhibitor. Observational literature on the association is conflicting, however important to understand for populations with a higher risk of medication-related adverse renal events. We aimed to systematically review the literature to summarize the association between sodium glucose cotransporter-2 inhibitor use and acute kidney injury, renal impairment, and acute renal failure in three at-risk groups: older people aged >65 years, people with heart failure, and people with reduced renal function. Methods Data Sources: A systematic search of Embase (1974–29/03/21) and PubMed (1946–29/03/21) was performed. Study Selection: RCTs and observational studies were included if they reported numbers of acute kidney injury or acute renal failure in people using sodium glucose cotransporter-2 inhibitors, compared to other diabetic therapies. Studies needed to report results by level of renal function, heart failure status, or age. Results Of 858 results, 6 studies were included. The absolute risk of acute kidney injury or acute renal failure was higher in people >65 years compared to those <65 years, higher in people with heart failure (vs. without), and higher in people with reduced kidney function (vs. preserved kidney function), but insufficient evidence to determine if the relative effect of sodium glucose cotransporter-2 inhibitors on this risk was similar for each group. Conclusion At risk cohorts are associated with a higher incidence of acute kidney problems in users of sodium glucose cotransporter-2 inhibitors.

The molluscan feeding structure is the radula, a chitinous membrane with teeth, which are highly adapted to the food and the substrate. In Polyplacophora and Patellogastropoda, the handling of hard ingesta can be facilitated by high content of chemical compounds containing Fe or Si in the tooth cusps. Other taxa, however, possess teeth that are less mineralized, even though animals have to avoid structural failure or high wear during feeding as well. Here, we investigated the gastropod Gastropteron rubrum, feeding on hard Foraminifera, diatoms and Porifera. Tooth morphologies and wear were documented by scanning electron microscopy and their mechanical properties were tested by nanoindentation. We determined, that gradients of hard- and stiffness run along each tooth, decreasing from cusp to basis. We also found, that inner lateral teeth are harder and stiffer than the outer ones. These findings allowed us to propose hypotheses about the radula-ingesta interaction. In search for the origins of the gradients, teeth were visualized using confocal laser scanning microscopy, to determine the degree of tanning, and analyzed with energy-dispersive X-ray spectroscopy, to test the elemental composition. We found that the mechanical gradients probably have their origin in the degree of tanning, as the teeth did not contain high proportions of metals or other minerals. However, in the tooth surfaces, which interact with the ingesta, high Si and Ca content was determined, which is likely an adaptation to reduce wear.

This editorial reviews the data to guide the discussion on whether or not to replace implantable cardioverter-defibrillator generator when the left ventricular ejection fraction has improved.

Atrial fibrillation (AF) provides extremel rapid excitation frequency in both atria, and induces several pathophysiological mechanisms by influencing each other to promote AF. This auto-enhancing feature is often described by the well-known concept “AF begets AF”. The cure of AF by creating pulmonary vein (PV) isolation may be able to reverse the spiral and be able to achieve reverse remodeling. Because of the initial focus on PV in the pathogenesis of AF, the effects of reverse remodeling after catheter ablation (CA) have been investigated on improvement of the left-sided cardiac system. On the other hand, tricuspid regurgitation (TR), the most neglected of all valvular diseases, is increasingly recognized as an important prognostic condition in heart failure patients. Previous reports, which tested the role of CA for AF patients with TR, have demonstrated that maintenance of SR provides reverse remodeling of the right-sided cardiac system, but have yet to prove whether this leads to improvement in patient prognosis. What is new and noteworthy about the report by Ukita et al. in this issue is that they found that TR improvement itself improves major event-free survival rate (incidence of heart failure hospitalization and all-cause mortality). However, several issues remain unresolved in their report. They observed a low AF recurrence rate in the TR-improved group, but did not address the possibility that AF suppression itself contributed to improve event-free survival rate. Further investigation is required to clarify whether TR improvement or maintenance of SR is the greater contributor to improved prognosis in AF.

A unique case of hybrid SEF-LGFMS with a EWSR1-CREB3L1 Fusion and robotic-assisted bronchoscopy in an 11-year-old Authors: Leore Lavin, MS1*, Nithya Krishnamurthy, BA1*, Chantal Spencer Grant, MD2, Kayleen Bailey, MD3Affiliations : 1Icahn School of Medicine at Mount Sinai, 2Mount Sinai Hospital Department of Pediatric Pulmonology, 3Mount Sinai Hospital Department of Pediatric Hematology-Oncology* Both authors contributed equally to this workCorresponding author: Leore Lavin,[email protected] East 98th St. New York, NY 10029. 646-709-8909.Running title: Rare pediatric SEF-LGFMS with EWSR1-CREB3L1 FusionKeywords: Robotic Bronchoscopy, Pediatric Oncology, SEF, LGFMS, EWSR1, CREB3L1Abbreviations:

Clinical Letter to the EditorConflict of interests’ statement: No potential conflict of interest is declared for any of the authorsTitle: Hemophagocytic lymphohistiocytosis in an adolescent withNLRP12 -related autoinflammatory disorder, a case report

6 month old child presented with complaints of recurrent respiratory infection, tachypnea and decreased air entry on right side . Xray chest AP view (Fig[1](#fig-cap-0001)) showed opaque right hemithorax with ipsilateral mediastinal shift and compensatory hyperinflation of left lung. Further evaluation with CECT thorax (Fig [2](#fig-cap-0002)) showed hypoplastic right lung with collapse consolidation and right sided mediastinal shift. Bifurcation of trachea was not seen with trachea continuing as left main bronchus with normal segmental division. Right bronchus appears to be arising from lower part of esophagus, precise communication was not demonstrated (Fig [3](#fig-cap-0003)). Conventional contrast (non ionic) esophagogram was performed and free flow of contrast was seen from lower third of esophagus to the right main bronchus (Fig [4](#fig-cap-0004)) confirming the diagnosis of Congenital bronchopulmonary foregut malformation (CBPFM)

Mixed Phenotype Acute Leukemia with BCL11B Copy Gain: What is the Best Strategy? Keri Toner MD1, 2, Reuven Schore MD1,2 and Jinjun Cheng MD PhD1,3,4*Center for Cancer and Blood Disorders, Children’s National Hospital, Washington, DCDepartment of Pediatrics, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, USADivision of Pathology and Laboratory Medicine, Children’s National Hospital, Washington, DCDepartments of Pathology and Pediatrics, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, USA

Introduction: The purpose of this study was to investigate whether patient, tumor and radiation therapy factors are associated with development of middle ear effusion (MEE) in nasopharyngeal carcinoma (NPC) patients. Methods: A retrospective review of NPC patients treated between January 2000 and June 2018 at Rabin Medical Center. Patient factors, tumor factors, radiation doses and radiation fields were collected and outlined if needed (middle ear, eustachian tube (ET), tensor veli palatini (TVP) and levator palatini (LVP) muscles), then analyzed and compared between patients with MEE and those without and between sides in patients with unilateral MEE. Results: 73 patients were enrolled. Most were males (71.2%) with advanced stage diseases (78%). At the time of diagnosis 14 patients (19.2%) presented with MEE and in 18 (24.6%) patients post radiation MEE was observed (15 ipsilateral to the tumor and 3 bilateral). Tumor stage, histology and laterality were not associated with development of MEE. Comparison of mean radiation field dosages including - gross target volume (GTV), clinical target volume (CTV) and patient target volume (PTV) showed no association with post radiation MEE. In addition, no difference was found in the radiation doses to the middle ear, ET or the LVP nor the TVP between ears with and without MEE. Conclusions: Postirradiation MEE remains a common adverse effect in NPC patients. Surprisingly, tumor stage, tumor laterality and histology were not associated with MEE. Similar findings were observed for total radiation doses and specific doses to the middle ear, ET and ET muscles.