bluuebunny/crossref_metadata_2025 huggingface dataset:
CD36 and ERK5 link dyslipidemia to apoptotic-like platelet procoagulant function
Authors: Yang Moua , Silverstein Roy L.
Published: July 2019

Purpose of review

Metabolic diseases, including dyslipidemia, diabetes mellitus, and chronic inflammation are risk factors for clinically significant thrombotic events. Thrombosis in these settings is multifaceted with coordinated mechanisms between platelet activation and the hemostatic pathways. This review focuses on recent advances in platelet procoagulant and apoptotic signaling with emphasis on the pathophysiologic mechanisms induced by platelet CD36 in dyslipidemia, and the key unaddressed questions relating to the field.

Recent findings

CD36 promotes platelet activation and increases the risk for thrombosis through signaling events. These include generation of reactive oxygen species, activation of redox-sensitive MAP kinase ERK5, and promotion of a pro-thrombotic phenotype. CD36 promotes phosphatidylserine externalization leading to a procoagulant function downstream from MAP kinase ERK5 that is separate from a pro-aggregatory function. Phosphatidylserine externalization requires maladaptive caspase activation, promotes assembly of the factor tenase and prothrombinase complex, and promotes fibrin formation. It is distinct from the canonical pathways mediating platelet procoagulant function by strong physiologic stimuli or by the platelet apoptotic-like Bak/Bax-mediated pathway for cellular clearance.

Understanding CD36 signaling in the context of dyslipidemia, or other metabolic diseases will identify important and novel signaling hubs that could be potential therapeutic targets for intervention without impacting hemostasis.
Rapid Whole Genome Sequencing Has Clinical Utility in Children in the PICU*
Authors: Sanford Erica F. , Clark Michelle M. , Farnaes Lauge , Williams Matthew R. , Perry James C. , Ingulli Elizabeth G. , Sweeney Nathaly M. , Doshi Ami , Gold Jeffrey J. , Briggs Benjamin , Bainbridge Matthew N. , Feddock Michele , Watkins Kelly , Chowdhury Shimul , Nahas Shareef A. , Dimmock David P. , Kingsmore Stephen F. , Coufal Nicole G.
Published: July 2019

Objectives:

Genetic disorders are a leading contributor to mortality in the neonatal ICU and PICU in the United States. Although individually rare, there are over 6,200 single-gene diseases, which may preclude a genetic diagnosis prior to ICU admission. Rapid whole genome sequencing is an emerging method of diagnosing genetic conditions in time to affect ICU management of neonates; however, its clinical utility has yet to be adequately demonstrated in critically ill children. This study evaluates next-generation sequencing in pediatric critical care.

Design:

Retrospective cohort study.

Setting:

Single-center PICU in a tertiary children’s hospital.

Patients:

Children 4 months to 18 years admitted to the PICU who were nominated between July 2016 and May 2018.

Interventions:

Rapid whole genome sequencing with targeted phenotype-driven analysis was performed on patients and their parents, when parental samples were available.

Measurements and Main Results:

A molecular diagnosis was made by rapid whole genome sequencing in 17 of 38 children (45%). In four of the 17 patients (24%), the genetic diagnoses led to a change in management while in the PICU, including genome-informed changes in pharmacotherapy and transition to palliative care. Nine of the 17 diagnosed children (53%) had no dysmorphic features or developmental delay. Eighty-two percent of diagnoses affected the clinical management of the patient and/or family after PICU discharge, including avoidance of biopsy, administration of factor replacement, and surveillance for disorder-related sequelae.

Conclusions:

This study demonstrates a retrospective evaluation for undiagnosed genetic disease in the PICU and clinical utility of rapid whole genome sequencing in a portion of critically ill children. Further studies are needed to identify PICU patients who will benefit from rapid whole genome sequencing early in PICU admission when the underlying etiology is unclear.
Sedative and Analgesic Drug Rotation Protocol in Critically Ill Children With Prolonged Sedation: Evaluation of Implementation and Efficacy to Reduce Withdrawal Syndrome*
Authors: Sanavia Eva , Mencía Santiago , Lafever Sarah N. , Solana María J. , Garcia Miriam , López-Herce Jesús
Published: July 2019

Objectives:

The first aim of this study was to assess the implementation of a sedative and analgesic drug rotation protocol in a PICU. The second aim was to analyze the incidence of withdrawal syndrome, drug doses, and time of sedative or analgesic drug infusion in children after the implementation of the new protocol.

Design:

Prospective observational study.

Setting:

PICU of a tertiary care hospital between June 2012 and June 2016.

Patients:

All patients between 1 month and 16 years old admitted to the PICU who received continuous IV infusion of sedative or analgesic drugs for more than 4 days were included in the study.

Interventions:

A sedative and analgesic drug rotation protocol was designed. The level of sedation, analgesia, and withdrawal syndrome were monitored with validated scales. The relationship between compliance with the protocol and the incidence of withdrawal syndrome was studied.

Measurements and Main Results:

One-hundred pediatric patients were included in the study. The protocol was followed properly in 35% of patients. Sixty-seven percent of the overall cohort presented with withdrawal syndrome. There was a lower incidence rate of withdrawal syndrome (34.3% vs 84.6%; p < 0.001), shorter PICU length of stay (median 16 vs 25 d; p = 0.003), less time of opioid infusion (median 5 vs 7 d for fentanyl; p = 0.004), benzodiazepines (median 5 vs 9 d; p = 0.001), and propofol (median 4 vs 8 d; p = 0.001) in the cohort of children in which the protocol was followed correctly.

Conclusions:

Our results show that compliance with the drug rotation protocol in critically ill children requiring prolonged sedation may reduce the appearance of withdrawal syndrome without increasing the risk of adverse effects. Furthermore, it may reduce the time of continuous IV infusions for most sedative and analgesic drugs and the length of stay in PICU.
Influence of Anxiety and Depression, Self-Rated Return-to-Work Problems, and Unemployment on the Outcome of Outpatient Rehabilitation After Shoulder Arthroscopy
Authors: Strube Patrick , Schöpe Tom , Hölzl Alexander , Brodt Steffen , Matziolis Georg , Zippelius Timo K.
Published: July 2019

Objective

The aim of this study was to research the influence of psychosocial confounders on outpatient rehabilitation after arthroscopic shoulder surgery.

Design

This retrospective study included patients who underwent such rehabilitation in a single center between January 2014 and October 2016. Shoulder function (Constant Shoulder Score) and pain (visual analog scale), improvements in these scores, and patient satisfaction were evaluated with regard to anxiety and depression (Hospital Anxiety and Depression Scale), self-rated return-to-work problems (Würzburg screening), and employment status.

Results

The analysis included 176 patients. The mean (SD) Constant Shoulder Score and visual analog scale improved from 53.9 (18) to 75.4 (16.5) and 4.6 (2.1) to 2.9 (2.4) cm, respectively. A total of 84.1% of the patients were satisfied with the outcome. Unemployed patients (P = 0.001) and Hospital Anxiety and Depression Scale–positive ones (P = 0.014) were less satisfied than their counterparts. Patients with a Würzburg screening–positive screening showed less improvement in pain (P = 0.015), function (P = 0.016), and satisfaction (P = 0.002) than those without. Unemployed reported more pain (P = 0.008) than employed patients when starting rehabilitation. At the end of rehabilitation, all psychosocial scores (Hospital Anxiety and Depression Scale, P = 0.002; Würzburg screening, P = 0.001; unemployment, P < 0.001) negatively influenced pain, Würzburg screening (P = 0.007), and unemployment (P = 0.008) function.

Conclusions

Because we identified psychosocial factors that influence the success of outpatient shoulder rehabilitation, rehabilitation setup should be adjusted in patients with such problems.
Magnetization Transfer Imaging Is Unaffected by Decreases in Renal Perfusion in Swine
Authors: Jiang Kai , Ferguson Christopher M. , Woollard John R. , Landes Vanessa L. , Krier James D. , Zhu Xiangyang , Nayak Krishna S. , Lerman Lilach O.
Published: July 2019

Objectives

Multiparametric renal magnetic resonance imaging (MRI), including diffusion-weighted imaging, magnetic resonance elastography, and magnetization transfer imaging (MTI), is valuable in the noninvasive assessment of renal fibrosis. However, hemodynamic changes in diseased kidneys may impede their ability to measure renal fibrosis. Because MTI assesses directly tissue content of macromolecules, we test the hypothesis that MTI would be insensitive to renal hemodynamic changes in swine kidneys with acute graded ischemia.

Materials and Methods

Seven domestic pigs underwent placement of an inflatable silicone cuff around the right renal artery to induce graded renal ischemia. Multiparametric MRI was performed at baseline, 50%, 75%, and 100% renal artery stenosis as well as reperfusion. Measurements included regional perfusion, R2*, apparent diffusion coefficient (ADC), stiffness, and magnetization transfer ratio (MTR) using arterial spin-labeled MRI, blood oxygenation–dependent MRI, diffusion-weighted imaging, magnetic resonance elastography, and MTI, respectively. Histology was performed to rule out renal fibrosis.

Results

During graded ischemia, decreases in renal perfusion were accompanied with elevated R2*, decreased ADC, and stiffness, whereas no statistically significant changes were observed in the MTR. No fibrosis was detected by histology. After release of the obstruction, renal perfusion showed only partial recovery, associated with return of kidney R2*, ADC, and stiffness to baseline levels, whereas cortical MTR decreased slightly.

Conclusions

Renal MTI is insensitive to decreases in renal perfusion and may offer reliable assessment of renal structural changes.
Postoperative Management After Total Pharyngolaryngectomy Using the Free Ileocolon Flap
Authors: Manrique Oscar J. , Sabbagh M. Diya , Kapoor Trishul , Ciudad Pedro , Chen Hung-Chi
Published: July 2019

Introduction

Management after total pharyngolaryngectomy with free ileocolon flaps can be challenging. Adequate postoperative surgical guidelines are essential to avoid complications. Factors, such as agitation, hypotension, or prolonged mechanical ventilation, might compromise final outcomes. Herein, we describe our experience in the early postoperative care of patients after total pharyngolaryngectomy with immediate reconstruction using the free ileocolon flap.

Methods

This is a retrospective review of all patients who underwent total pharyngolaryngectomy and immediate reconstruction using the free Ileocolon flap. Demographics, etiology of resection, neoadjuvant therapy, surgical time, method of sedation, postoperative use of vasopressors, length of intensive care unit (ICU) stay, time of discontinuation of mechanical ventilation, and complications were recorded and analyzed.

Results

Between 2010 and 2015, a total of 34 patients underwent total pharyngolaryngectomy and immediate reconstruction using the free Ileocolon flap. The most common cause of total pharyngolaryngectomy was cancer. Twenty-eight patients had neoadjuvant therapy (radiation). The average surgical time was 11.5 hours (range, 8–14.5 hours), average length of ICU stay was 3 days (range, 2–15 days) with an average time for mechanical ventilation cessation of 3 days (range, 1–20 days). Midazolam and dexmedetomidine were the most common sedatives used during surgery and in the ICU period. Three patients required vasopressors due to hypotension, 2 had unplanned self-extubation from the tracheostomy site, 2 experienced postoperative bleeding, 1 had pneumonia, 4 required unplanned return to the operating room, 2 had partial flap loss, and 1 had complete flap loss.

Conclusions

Overall, a majority of patients recovered well postoperatively with minimal complications and low rate of reoperation. Our research provides a foundation to develop a risk-stratified approach to determine the need for an ICU admission or early transfer to floor care.
Difficult Airway Identification and Management in Patients With Moebius Syndrome
Authors: Telich-Tarriba Jose E. , Guevara-Valmaña Osvaldo , Ruiz-Beltran Sandra , Aguiar-Rosas Sebastian , Cardenas-Mejia Alexander
Published: July 2019

Background:

Airway management in patients with Moebius syndrome can be difficult due to the presence of orofacial malformations. This paper aims to present our institution's experience in the evaluation and management of the upper airway in patients with Moebius syndrome

Methodology:

The authors performed a retrospective study including every patient with Moebius syndrome submitted to surgery between 2012 and 2017. Difficult airway was defined as one requiring more than 2 attempts to achieve endotracheal tube placement, a Mallampati score equal or above III, a Cormack score equal or above III, or need of a fiberscope during intubation.

Results:

Fifty-one patients were included, (39.3% males) requiring 172 procedures. Each patient required an average of 3.37 ± 1.94 anesthetic events. Average Body Mass Index (BMI) was 20.07 ± 6.06, 45.1% were overweight or obese. Four patients (7%) were considered as having a potentially difficult airway. Endotracheal intubation was achieved in all patients; 38 patients were intubated in a single attempt, while the rest were successfully intubated on a second try. Fiberscope was not necessary. Univariate analysis showed that overweight/obese patients were more likely to be considered as having a difficult airway (P = 0.03).

Conclusion:

Intubation can be difficult in patients with Moebius syndrome, but failure is rare. Overweight or obese patients are at risk of presenting a difficult airway. An extensive preoperative evaluation and adequate communication between the members of the multidisciplinary team in charge of these patients is paramount.
Lipoma Rarely Involving Multiple Suprahyoid Fascial Spaces: A Large Lesion Removed With Bimanual Transcervical Finger Dissection Using a “Push–Pull Down” Maneuver
Authors: Imai Tomoaki , Nakazawa Mitsuhiro , Uzawa Narikazu
Published: July 2019

Lipoma involving multiple fascial spaces is extremely rare and poses a challenge to surgeons using less invasive procedures. Although blunt dissection using a finger is often used in excisional surgeries as a supplementary maneuver, few cases have been described using the bimanual technique for the removal of these extensive lesions.

Herein, the authors present a large lipoma of the oral floor extending to the unilateral submandibular and parapharyngeal spaces, which was excised only via a submandibular approach using the “push–pull down” maneuver, blunt finger dissection of the tumor with counter-pushing on the overlying mucosa to the submandibular direction facilitated tumor separation. The tumor was consequently pulled down and removed without an additional intraoral incision.

Based on accurate preoperative examinations, this maneuver, provides a less invasive surgery for well-encapsulated benign tumors involving multiple fascial spaces.
Bilateral Sagittal Split Osteotomy: Description of Surgical Technique to Complement the Procedural Cognition Simulation in the Craniofacial Interactive Virtual Assistant-Professional Edition
Authors: Rodriguez Eduardo D. , Plana Natalie M. , Diaz-Siso Jesus Rodrigo , Flores Roberto L.
Published: July 2019

Operative experience for the contemporary trainee has become exceedingly more challenging in the setting of more stringent hospital regulations. Surgical training is thus shifting toward more self-directed, independent learning to maximize operative opportunities as they become available; yet, this can prove difficult for complex surgeries like craniofacial procedures. The intricate anatomy and fine reconstructive techniques employed cannot be readily depicted onto a two-dimensional page. To address this educational gap, the Craniofacial Interactive Virtual Assistant-Pro Edition (CIVA-Pro) was developed as a web-based surgical simulator to aid learners with conceptualizing the surgical principles utilized in these cases. The current work reviews the Bilateral Sagittal Split Osteotomy module of CIVA-Pro, providing detailed narratives for each chapter with expert commentary on broadened indications and future directions.
The Characteristics of Lower Eyelid Reverse Ptosis After Reconstruction of Orbital Floor Wall Fracture Using Transconjunctival Approach
Authors: Park Jinhwan , Kim Seungheon , Baek Sehyun
Published: July 2019

The purpose of this study was to evaluate the changes of lower eyelid position and the incidence of reverse ptosis after reconstruction of orbital floor fracture. The authors retrospectively reviewed the clinical records of patients who received reconstruction of orbital floor wall fractures between 2014 and 2017. Digital photographs were taken preoperatively, and at 1 week, 1 month, and 3 months postoperatively. Main measurements were marginal reflex distance 2 (MRD 2), which was measured by analyzing the digital photographs using ImageJ software. Among 264 patients with orbital wall fracture, The authors enrolled 32 eyes (18 right eyes and 14 left eyes) of 32 patients (21 males and 11 females) with a mean age of 32.1 years (range, 16–57 years). 7 (21.9%) of 32 patients had reverse ptosis at postoperative 3 months. When MRD2 of affected eye was compared based on the fellow eye, 7 patients with reversed ptosis showed a definite MRD 2 decrease (>1 mm) of affected eye from postoperative 1 month. Age showed a significant negative correlation with the difference of MRD2 between affected and fellow eye at postoperative 1 and 3 months (r = −0.378, P = 0.033 and r = −0.372, P = 0.036, respectively). Postoperative complications were not observed in all patients. The transconjunctival access in orbital floor wall surgery is a safe and useful surgical approach. However, some may have a reverse ptosis postoperatively, especially older patients. Reverse ptosis is a major clinical finding that should not be overlooked in post-operative follow-up.
Surgical Management of Large Odontogenic Cysts of the Mandible
Authors: Bonavolontà Paola , Dell’Aversana Orabona Giovanni , Friscia Marco , Sani Lorenzo , Abbate Vincenzo , Iaconetta Giorgio , Califano Luigi
Published: July 2019

Introduction:

Odontogenic cysts are defined as those cysts that arise from odontogenic epithelium and occur in the tooth-bearing regions of the jaws. Cystectomy, marsupialization, or decompression of odontogenic cyst are the most common treatments proposed for this pathology. The aim of this study is to retrospectively evaluate the result of decompression based on the volumetric reduction of the cystic cavity and new bone formation by cone beam computerized tomography (CBCT).

Methods:

The 16 patients affected by a large odontogenic mandibular cyst were enrolled in the study. All the patients underwent a surgical decompression of the cyst followed by the enucleation after a follow-up ranging from 6 to 9 months according to the volume's reduction and new bone formation. All the patients were evaluated with a CBCT before and after the surgical decompression to measure and analyze the percentage of reduction of the cystic volume before proceeding with the enucleation.

Results:

The decompression of the cyst showed a reduction of the cystic volume ranging from 38.2% to 54.4% proportionally to the treatment duration. The highest percentage of volume reduction observed was 54.4% in 1 patient followed-up for 9 months, before the surgical enucleation.

Conclusion:

In our experience, the decompression seems to be the most suitable technique for the primary treatment of large odontogenic cyst of the jaws followed by the enucleation after 6 to 9 months. The CBCT is an objective method to evaluate the cystic volume reduction after the decompression and helps the surgeon with the surgical planning.
Changes in Vital Signs During Epileptic and Psychogenic Nonepileptic Attacks: A Video-EEG Study
Authors: Badry Reda
Published: July 2019

Introduction:

Epilepsy is a common disorder. Psychogenic nonepileptic attack (PNEA) is one of the epilepsy mimics. Video EEG is still the gold standard tool that differentiates between epileptic seizures (ES) and PNEA. Oxygen saturation (SaO2) and ictal vital signs, including heart rate (HR), respiration rate (RR), body temperature, systolic blood pressure (SBP), and diastolic blood pressure show crucial changes during ES and PNEA.

Purpose:

To analyze, compare, and find relationships of changes in ictal vital sign during ES and PNEA.

Methods:

Ninety-four adults had video EEG monitoring. SaO2, HR, RR, body temperature, SBP, and diastolic blood pressure were obtained at baseline and during the attacks.

Results:

The 49 patients with ES and 45 patients with PNEA had nearly similar baseline vital sign. Epileptic seizures yielded a higher ictal HR (p = 0.003) and lower ictal SaO2 (p = 0.04) than PNEA. Ictal RR and ictal SBP of patients with PNEA were higher than those of patients with ES (p = 0.02 and P = 0.04, respectively). For ES, ictal HR inversely correlated with ictal SaO2 (P = 0.003). In PNEA, ictal HR directly correlated with ictal SBP (P = <0.005).

Conclusions:

There are statistically significant differences between pre-ictal and ictal SaO2, HR, SBP, and diastolic blood pressure in both ES and PNEA groups. The inverse relationship between ictal SaO2 and ictal HR in ES suggests severe cardiorespiratory dysfunction. The significantly elevated ictal HR, ictal RR, and ictal SBP during PNEA demonstrates the risk of those attacks if not stopped.
Connections between the cell cycle, cell adhesion and the cytoskeleton
Authors: Jones Matthew C. , Zha Junzhe , Humphries Martin J.
Published: July 2019

Cell division, the purpose of which is to enable cell replication, and in particular to distribute complete, accurate copies of genetic material to daughter cells, is essential for the propagation of life. At a morphological level, division not only necessitates duplication of cellular structures, but it also relies on polar segregation of this material followed by physical scission of the parent cell. For these fundamental changes in cell shape and positioning to be achieved, mechanisms are required to link the cell cycle to the modulation of cytoarchitecture. Outside of mitosis, the three main cytoskeletal networks not only endow cells with a physical cytoplasmic skeleton, but they also provide a mechanism for spatio-temporal sensing via integrin-associated adhesion complexes and site-directed delivery of cargoes. During mitosis, some interphase functions are retained, but the architecture of the cytoskeleton changes dramatically, and there is a need to generate a mitotic spindle for chromosome segregation. An economical solution is to re-use existing cytoskeletal molecules: transcellular actin stress fibres remodel to create a rigid cortex and a cytokinetic furrow, while unipolar radial microtubules become the primary components of the bipolar spindle. This remodelling implies the existence of specific mechanisms that link the cell-cycle machinery to the control of adhesion and the cytoskeleton. In this article, we review the intimate three-way connection between microenvironmental sensing, adhesion signalling and cell proliferation, particularly in the contexts of normal growth control and aberrant tumour progression. As the morphological changes that occur during mitosis are ancient, the mechanisms linking the cell cycle to the cytoskeleton/adhesion signalling network are likely to be primordial in nature and we discuss recent advances that have elucidated elements of this link. A particular focus is the connection between CDK1 and cell adhesion.

This article is part of a discussion meeting issue ‘Forces in cancer: interdisciplinary approaches in tumour mechanobiology’.
Validation of the Seegene RV15 multiplex PCR for the detection of influenza A subtypes and influenza B lineages during national influenza surveillance in hospitalized adults
Authors: LeBlanc J. J. , ElSherif M. , Mulpuru S. , Warhuus M. , Ambrose A. , Andrew M. , Boivin G. , Bowie W. , Chit A. , Dos Santos G. , Green K. , Halperin S. A. , Hatchette T. F. , Ibarguchi B. , Johnstone J. , Katz K. , Langley J. M. , Lagacé-Wiens P. , Loeb M. , Lund A. , MacKinnon-Cameron D. , McCarthy A. , McElhaney J. E. , McGeer A. , Poirier A. , Powis J. , Richardson D. , Semret M. , Shinde V. , Smyth D. , Trottier S. , Valiquette L. , Webster D. , Ye L. , McNeil S. A.
Published: July 2019

Background. The Serious Outcomes Surveillance Network of the Canadian Immunization Research Network (CIRN SOS) has been performing active influenza surveillance since 2009 (ClinicalTrials.gov identifier: NCT01517191). Influenza A and B viruses are identified and characterized using real-time reverse-transcriptase polymerase chain reaction (RT-PCR), and multiplex testing has been performed on a subset of patients to identify other respiratory virus aetiologies. Since both methods can identify influenza A and B, a direct comparison was performed.

Methods. Validated real-time RT-PCRs from the World Health Organization (WHO) to identify influenza A and B viruses, characterize influenza A viruses into the H1N1 or H3N2 subtypes and describe influenza B viruses belonging to the Yamagata or Victoria lineages. In a subset of patients, the Seeplex RV15 One-Step ACE Detection assay (RV15) kit was also used for the detection of other respiratory viruses.

Results. In total, 1111 nasopharyngeal swabs were tested by RV15 and real-time RT-PCRs for influenza A and B identification and characterization. For influenza A, RV15 showed 98.0 % sensitivity, 100 % specificity and 99.7 % accuracy. The performance characteristics of RV15 were similar for influenza A subtypes H1N1 and H3N2. For influenza B, RV15 had 99.2 % sensitivity, 100 % specificity and 99.8 % accuracy, with similar assay performance being shown for both the Yamagata and Victoria lineages.

Conclusions. Overall, the detection of circulating subtypes of influenza A and lineages of influenza B by RV15 was similar to detection by real-time RT-PCR. Multiplex testing with RV15 allows for a more comprehensive respiratory virus surveillance in hospitalized adults, without significantly compromising the reliability of influenza A or B virus detection.
Tumor cell-organized fibronectin is required to maintain a dormant breast cancer population
Authors: Barney Lauren E. , Hall Christopher L. , Schwartz Alyssa D. , Parks Akia N. , Sparages Christopher , Galarza Sualyneth , Platt Manu O. , Mercurio Arthur M. , Peyton Shelly R.
Published: July 2019

Tumors can undergo long periods of dormancy, with cancer cells entering a largely quiescent, non-proliferative state before reactivation and outgrowth. For a patient, these post-remission tumors are often drug resistant and highly aggressive, resulting in poor prognosis. To understand the role of the extracellular matrix (ECM) in regulating tumor dormancy, we created anin vitrocell culture system that combines carefully controlled ECM substrates with nutrient deprivation to observe entranceintoand exitfromdormancy with live imaging. We saw that cell populations capable of surviving entrance into long-term dormancy were heterogeneous, containing quiescent, cell cycle arrested, and actively proliferating cells. Cell populations that endured extended periods of serum-deprivation-induced dormancy formed an organized, fibrillar fibronectin matrix via αvβ3and α5β1integrin adhesion, ROCK-generated tension, and TGFβ2 stimulation. We surmised that the fibronectin matrix was primarily a mediator of cell survival, not proliferation, during the serum-deprivation stress, bacause cancer cell outgrowth after dormancy required MMP-2-mediated fibronectin degradation. Given the difficulty of animal models in observing entrance and exit from dormancy in real-time, we propose this approach as a new,in vitromethod to study factors important in regulating dormancy, and we used it here to elucidate a role for fibronectin deposition and MMP activation.
Chemogenetic silencing of neurons in the mouse anterior cingulate area modulates neuronal activity and functional connectivity
Authors: Peeters Lore M. , Hinz Rukun , Detrez Jan R. , Missault Stephan , De Vos Winnok H. , Verhoye Marleen , Van der Linden Annemie , Keliris Georgios A.
Published: July 2019

The anterior cingulate area (ACA) is an integral part of the prefrontal cortex in mice and has been implicated in several cognitive functions. Previous anatomical and functional imaging studies demonstrated that the ACA is highly interconnected with numerous brain regions acting as a hub region in functional networks. However, the importance of the ACA in regulating functional network activity and connectivity remains to be elucidated. Recently developed neuromodulatory techniques, such as Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) allow for precise control of neuronal activity. In this study, we used an inhibitory kappa-opioid receptor DREADDs (KORD) to temporally inhibit neuronal firing in the right ACA of mice and assessed functional network activity and connectivity using non-invasive functional MRI. We demonstrated that KORD-induced inhibition of the right ACA induced blood oxygenation-level dependent (BOLD) signal decreases and increases in connected brain regions throughout of hemispheres. Furthermore, these modulations in neuronal activity were associated with decreased intra- and interhemispheric functional connectivity. These results demonstrate that the combination of the DREADD technology and non-invasive functional imaging methods is a valuable tool for unraveling the underlying mechanisms of network function and dysfunction.
Systematic phenomics analysis of ASD-associated genes reveals shared functions and parallel networks underlying reversible impairments in habituation learning
Authors: McDiarmid Troy A. , Belmadani Manuel , Liang Joseph , Meili Fabian , Mathews Eleanor A. , Mullen Gregory P. , Rand James B. , Mizumoto Kota , Haas Kurt , Pavlidis Paul , Rankin Catharine H.
Published: July 2019

A major challenge facing the genetics of Autism Spectrum Disorders (ASD) is the large and growing number of candidate risk genes and gene variants of unknown functional significance. Here, we used Caenorhabditis elegans to systematically functionally characterize ASD-associated genes in vivo. Using our custom machine vision system we quantified 26 phenotypes spanning morphology, locomotion, tactile sensitivity, and habituation learning in 87 strains each carrying a mutation in an ortholog of an ASD-associated gene. We identified hundreds of novel genotype-phenotype relationships ranging from severe developmental delays and uncoordinated movement to subtle deficits in sensory and learning behaviors. We clustered genes by similarity in phenomic profiles and used epistasis analysis to discover parallel networks centered on CHD8•chd-7 and NLGN3•nlg-1 that underlie mechanosensory hyper-responsivity and impaired habituation learning. We then leveraged our data for in vivo functional assays to gauge missense variant effect. Expression of wild-type NLG-1 in nlg-1 mutant C. elegans rescued their sensory and learning impairments. Testing the rescuing ability of all conserved ASD-associated neuroligin variants revealed varied partial loss-of-function despite proper subcellular localization. Finally, we used CRISPR-Cas9 auxin inducible degradation to determine that phenotypic abnormalities caused by developmental loss of NLG-1 can be reversed by adult expression. This work charts the phenotypic landscape of ASD-associated genes, offers novel in vivo variant functional assays, and potential therapeutic targets for ASD.
Wide-field time-gated SPAD imager for phasor-based FLIM applications
Authors: Ulku Arin , Ardelean Andrei , Antolovic Michel , Weiss Shimon , Charbon Edoardo , Bruschini Claudio , Michalet Xavier
Published: July 2019

We describe the performance of a new wide area time-gated single-photon avalanche diode (SPAD) array for phasor-FLIM, exploring the effect of gate length, gate number and signal intensity on the measured lifetime accuracy and precision. We conclude that the detector functions essentially as an ideal shot noise limited sensor and is capable of video rate FLIM measurement. The phasor approach used in this work appears ideally suited to handle the large amount of data generated by this type of very large sensor (512×512 pixels), even in the case of small number of gates and limited photon budget.
Genome-wide scan identifies novel genetic loci regulating salivary metabolite levels
Authors: Nag Abhishek , Kurushima Yuko , Bowyer Ruth C. E. , Wells Philippa M. , Weiss Stefan , Pietzner Maik , Kocher Thomas , Raffler Johannes , Völker Uwe , Mangino Massimo , Spector Timothy D. , Milburn Michael V. , Kastenmüller Gabi , Mohney Robert P. , Suhre Karsten , Menni Cristina , Steves Claire J.
Published: July 2019

Saliva, as a biofluid, is inexpensive and non-invasive to obtain, and provides a vital tool to investigate oral health and its interaction with systemic health conditions. There is growing interest in salivary biomarkers for systemic diseases, notably cardiovascular disease. Whereas hundreds of genetic loci have been shown to be involved in the regulation of blood metabolites leading to unprecedented insights into the pathogenesis of complex human diseases, little is known about the impact of host genetics on salivary metabolites. Here we report the first genome-wide association study exploring 476 salivary metabolites in 1,419 subjects of European ancestry from the TwinsUK cohort (discovery phase). A total of 14 salivary metabolites were significantly associated (p<10−10) with genetic variants that mapped to 11 distinct loci, most of which replicated in the Study of Health in Pomerania (SHIP-2) cohort. Interestingly, while only a limited number of the loci that are known to regulate blood metabolites were also associated with salivary metabolites in our study, we identified several novel saliva-specific locus-metabolite associations, including associations for theAGMAT(with the metabolites 4-guanidinobutanoate and beta-guanidinopropanoate),ATP13A5(with the metabolite creatinine) andDPYS(with the metabolites 3-ureidopropionate and 3-ureidoisobutyrate) loci. Our study suggests that there are biological pathways which are specific to the regulation of the salivary metabolome. In addition, some of our findings may have clinical relevance, such as the utility of the pyrimidine (uracil) degradation metabolites in predicting 5-fluorouracil toxicity and the role of the agmatine pathway metabolites as biomarkers of oral health.
Development of a Surface Programmable Activation Receptor system (SPAR): A living cell biosensor for rapid pathogen detection
Authors: Kittle Joseph D. , Lwande Joel S. , Williams M. Russell , Brody Richard , Frenchmeyer Melissa , Hua Jiangzhou , Liang Shengwen , McQuaid Kyle , Mo Min , Neese Allison , Tang Yuanyuan , Vedamoorthy Srikanth , Zeng Lingchun , Zupancic Thomas , McBrairty Charles
Published: July 2019

Efficient pathogen detection is essential for the successful treatment and prevention of infectious disease; however, current methods are often too time intensive to be clinically relevant in cases requiring immediate intervention. We have developed a Surface Programmable Activation Receptor (SPAR) diagnostic platform comprised of universal biosensor cells engineered for use in combination with custom or commercial antibodies to achieve rapid and sensitive pathogen detection. SPAR cells are stably transfected Jurkat T cells designed to constitutively express a modified T cell mouse FcγRI receptor on the cell surface and a high level of the luminescent reporter protein aequorin in the cytoplasm. The modified mFcγRI-CD3ζ receptor protein binds with high affinity to the Fc region of any full-length mouse IgG2a and some IgG2 antibodies: this allows customized target detection via the selection of specific antibodies. T-cell receptor aggregation in response to target antigen binding results in signal transduction which, when amplified via the endogenous T cell signal cascade, triggers the rapid intracellular release of calcium. Increased Ca2+ concentrations activate the expressed reporter protein aequorin resulting in the immediate emission of detectable light. Testing demonstrates the accurate and specific detection of numerous targets including P. aeruginosa, E. coli O111, and E. coli O157. We report that the SPAR biosensor cell platform is a reliable pathogen detection method that enables the rapid identification of bacterial causative agents using standard laboratory instrumentation. The technology lends itself to the development of efficient point-of-care testing and may aid in the implementation of effective and pathogen-specific clinical therapies.
Chronic paroxetine blunts stress response and normalizes adverse behavioral effects seen acutely in nulliparous rats
Authors: Tuckute Greta , Overgaard Agnete , Frokjaer Vibe G.
Published: July 2019

Selective serotonin reuptake inhibitors (SSRI) are widely used antidepressants and their effect is partly mediated by restoring stress axis dynamics, which may depend on sex and hormonal states. In the present study, we investigate the effect of daily injections of the SSRI paroxetine (5 mg/kg s.c.) on swim stress-induced corticosterone (CORT) response, depressive-like behavior (forced swim test) and anxiety-like behavior (open field test) in nulliparous Sprague Dawley rats. Data were acquired after either 1-3 (acute PXT) or 11-13 (chronic PXT) injections. We found that chronic, but not acute, paroxetine blunted the swim stress-induced CORT response. We observed an increase in depressive-like and anxiety-like behavior following acute PXT, and a normalization after chronic PXT treatment. Intriguingly, our findings of rapid recovery from adverse SSRI effects differ from corresponding studies performed by our group in postpartum rats. Thus, the study emphasizes that mechanisms of action and efficacy of SSRIs differ according to reproductive states, which if translated to humans may inform treatment strategies, beyond SSRIs alone, for hormone transition related depressive states.
Multistability and regime shifts in microbial communities explained by competition for essential nutrients
Authors: Dubinkina Veronika , Fridman Yulia , Pandey Parth Pratim , Maslov Sergei
Published: July 2019

Microbial communities routinely have several possible species compositions or community states observed for the same environmental parameters. Changes in these parameters can trigger abrupt and persistent transitions (regime shifts) between such community states. Yet little is known about the main determinants and mechanisms of multistability in microbial communities. Here we introduce and study a resource-explicit model in which microbes compete for two types of essential nutrients. We adapt game-theoretical methods of the stable matching problem to identify all possible species compositions of a microbial community. We then classify them by their resilience against three types of perturbations: fluctuations in nutrient supply, invasions by new species, and small changes of abundances of existing ones. We observe multistability and explore an intricate network of regime shifts between stable states in our model. Our results suggest that multistability requires microbial species to have different stoichiometries of essential nutrients. We also find that balanced nutrient supply promote multistability and species diversity yet make individual community states less stable.
Convergent metatarsal fusion in jerboas and chickens is mediated by similarities and differences in the patterns of osteoblast and osteoclast activities
Authors: Gutierrez Haydee L. , Tsutsumi Rio , Moore Talia Y. , Cooper Kimberly L.
Published: July 2019

The extraordinary malleability of the vertebrate limb supports a variety of locomotor functions including running and leaping in cursorial and saltatorial species. In many of these animals, the metatarsals and/or metacarpals are disproportionately elongated to increase stride length and fused into a single larger element, likely to resist fracture due to increased ground reaction forces. Despite the fact that metapodial fusion evolved convergently in modern birds, ungulates, and jerboas, the developmental basis has only been explored in chickens, which diverged from the mammalian lineage approximately 300 million years ago. Here, we use the lesser Egyptian jerboa, Jaculus jaculus, to understand the cellular processes that unite three distinct metatarsal elements into a single cannon bone in a mammal, and we revisit the developing chicken to assess similarities and differences in the localization of osteoblast and osteoclast activities. In both species, adjacent metatarsals align along flat surfaces, osteoblasts cross the periosteal membrane to unite the three elements in a single circumference, and osteoclasts resorb bone at the interfaces leaving a single marrow cavity. However, although spatial and temporal partitioning of osteoblast and osteoclast activities reshape three bones into one in both species, the localization of osteoclasts is distinct. While osteoclasts are uniformly distributed throughout the endosteum of chicken metatarsals, these catabolic cells are highly localized to resorb bone at the interfaces of neighboring jerboa metatarsals. Each species therefore provides an opportunity to better understand the mechanisms that partition osteoblasts and osteoclasts to alter the shape of bone during development and evolution.
The plant pathogenPectobacterium atrosepticumcontains a functional formate hydrogenlyase-2 complex
Authors: Finney Alexander J. , Lowden Rebecca , Fleszar Michal , Albareda Marta , Coulthurst Sarah J. , Sargent Frank
Published: July 2019

Pectobacterium atrosepticumSCRI1043 is a phytopathogenic gram-negative enterobacterium. Genomic analysis has identified that genes required for both respiration and fermentation are expressed under anaerobic conditions. One set of anaerobically expressed genes is predicted to encode an important but poorly-understood membrane-bound enzyme termed formate hydrogenlyase-2 (FHL-2), which has fascinating evolutionary links to the mitochondrial NADH dehydrogenase (Complex I). In this work, molecular genetic and biochemical approaches were taken to establish that FHL-2 is fully functional inP. atrosepticumand is the major source of molecular hydrogen gas generated by this bacterium. The FHL-2 complex was shown to comprise a rare example of an active [NiFe]-hydrogenase-4 (Hyd-4) isoenzyme, itself linked to an unusual selenium-free formate dehydrogenase in the final complex. In addition, further genetic dissection of the genes encoding the predicted membrane domain of FHL-2 established surprisingly that the majority of genes encoding this domain are not required for physiological hydrogen production activity. Overall, this study presentsP. atrosepticumas a new model bacterial system for understanding anaerobic formate and hydrogen metabolism in general, and FHL-2 function and structure in particular.

Significance Statement

Pectobacterium atrospecticumcontains the genes for the formate hydrogenlyase-2 enzyme, considered the ancient progenitor of mitochondrial respiratory Complex I. In this study, the harnessing ofP. atrosepticumas a new model system for understanding bacterial hydrogen metabolism has accelerated new knowledge in FHL-2 and its component parts. Importantly, those component parts include an unusual selenium-free formate dehydrogenase and a complicated [NiFe]-hydrogenase-4 with a large membrane domain. FHL-2 is established as the major source of molecular hydrogen produced under anaerobic conditions byP. atrospectium, however surprisingly some components of the membrane domain were not essential for this activity.
Classifying disease-associated variants using measures of protein activity and stability
Authors: Jepsen Michael Maglegaard , Fowler Douglas M. , Hartmann-Petersen Rasmus , Stein Amelie , Lindorff-Larsen Kresten
Published: July 2019

Decreased cost of human exome and genome sequencing provides new opportunities for diagnosing genetic disorders, but we need better and more robust methods for interpreting sequencing results including determining whether and by which mechanism a specific missense variants may be pathogenic. Using the protein PTEN (phosphatase and tensin homolog) as an example, we show how recent developments in both experiments and computational modelling can be used to determine whether a missense variant is likely to be pathogenic. One approach relies on multiplexed experiments that enable determination of the effect of all possible individual missense variants in a cellular assay. Another approach is to use computational methods to predict variant effects. We compare two different multiplexed experiments and two computational methods to classify variant effects in PTEN. We distinguish between methods that focus on effects on protein stability and protein-specific methods that are more directly related to enzyme activity. Our results on PTEN suggest that ~60% of pathogenic variants cause loss of function because they destabilise the folded protein which is subsequently degraded. Methods that quantify a broader range of effects on PTEN activity perform better at predicting variant effects. Either experimental method performs better than the corresponding computational predictions, so that e.g. experiments that probe cellular abundance perform better at identifying pathogenic variants than predictions of thermodynamic stability. Our results suggest that loss of stability of PTEN is a key driver for disease, and we hypothesize that experiments and prediction methods that probe protein stability can be used to find variants with similar mechanisms in other genes.
Brain Regional Gene Expression Network Analysis Identifies Unique Interactions Between Chronic Ethanol Exposure and Consumption
Authors: Smith M.L. , Lopez M.F. , Wolen A.R. , Becker H.C. , Miles M.F.
Published: July 2019

Progressive increases in ethanol consumption is a hallmark of alcohol use disorder (AUD). Persistent changes in brain gene expression are hypothesized to underlie the altered neural signaling producing abusive consumption in AUD. To identify brain regional gene expression networks contributing to progressive ethanol consumption, we performed microarray and scale-free network analysis of expression responses in a C57BL/6J mouse model utilizing chronic intermittent ethanol by vapor chamber (CIE) in combination with limited access oral ethanol consumption. This model has previously been shown to produce long-lasting increased ethanol consumption, particularly when combining oral ethanol access with repeated cycles of intermittent vapor exposure. The interaction of CIE and oral consumption was studied by expression profiling and network analysis in medial prefrontal cortex, nucleus accumbens, hippocampus, bed nucleus of the stria terminalis, and central nucleus of the amygdala. Brain region expression networks were analyzed for ethanol-responsive gene expression, correlation with ethanol consumption and functional content using extensive bioinformatics studies. In all brain-regions studied the largest number of changes in gene expression were seen when comparing ethanol naïve mice to those exposed to CIE and drinking. In the prefrontal cortex, however, unique patterns of gene expression were seen compared to other brain-regions. Network analysis identified modules of co-expressed genes in all brain regions. The prefrontal cortex and nucleus accumbens showed the greatest number of modules with significant correlation to drinking behavior. Across brain-regions, however, many modules with strong correlations to drinking, both baseline intake and amount consumed after CIE, showed functional enrichment for synaptic transmission and synaptic plasticity.