Shock

Beneficial Effects of Ivabradine on Post-Resuscitation Myocardial Dysfunction In A Porcine Model of Cardiac Arrest Background: Ivabradine selectively inhibits the If current, reducing the heart rate and protecting against myocardial ischemia/reperfusion injury. We investigated the effects of ivabradine on post-resuscitation myocardial function in a porcine model of cardiopulmonary resuscitation. Methods and Results: Ventricular fibrillation was induced and untreated for 8 minutes while defibrillation was attempted after 6 minutes of cardiopulmonary resuscitation in anesthetized domestic swine. Then the animals were randomized into ivabradine and placebo groups (n = 5 each). Ivabradine and saline were administered at the same volume 5 minutes after ROSC (Return of Spontaneous Circulation), followed by continuous intravenous infusion at 0.5 mg/kg for 480 minutes. Hemodynamic parameters were continuously recorded. Myocardial function was assessed by echocardiography at baseline and at 60, 120, 240, 480 minutes and 24 hours after resuscitation. The serum levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI) were measured by commercial enzyme-linked immunosorbent assay kits. Animals were killed 24 hours after resuscitation, and all myocardial tissue was removed for histopathological analysis. The heart rate was significantly reduced from 1 hour after resuscitation in the ivabradine group (all p < 0.05). The post-resuscitation mitral E/A and E/e′ velocity ratios and left ventricular ejection fraction were significantly better in the ivabradine than placebo group (p < 0.05). The serum levels of myocardial injury biomarkers (NT-proBNP, cTnI) and the myocardial biopsy scores were significantly lower in the ivabradine than placebo group (p < 0.05). Neurological deficit scores were lower in the IVA group at PR 24 hours (p < 0.05). Conclusions: Ivabradine improved post-resuscitation myocardial dysfunction, myocardial injury and post-resuscitation cerebral function, and also slowed the heart rate in this porcine model. Address reprint requests to Min Yang, MD, PhD, The 2nd Department of Intensive Care Unit, No. 2 Hospital Affiliated to Anhui Medical University, Furong Road 678, Hefei, China, 230032. E-mail: 512130761@qq.com Received 4 April, 2019 Revised 25 April, 2019 Accepted 27 June, 2019 Source of funding: This study was supported by the National Natural Science Foundation of China (No. 81601661), Natural Science Foundation of Anhui Province of China (No. 1608085MH195), and Science Foundation for Post-doctoral Researchers in Anhui Province of China (No. 2016B140). Disclosure: The authors have no conflicts of interest to declare. This is an open access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://creativecommons.org/licenses/by-nc-nd/4.0 © 2019 by the Shock Society

Early Maladaptive Cardiovascular Responses are Associated with Mortality In A Porcine Model of Hemorrhagic Shock Background: Hemorrhage is a leading cause of death on the battlefield. Current methods for predicting hemodynamic deterioration during hemorrhage are of limited accuracy and practicality. During a study of the effects of remote ischemic preconditioning in pigs that underwent hemorrhage, we noticed arrhythmias among all pigs that died before the end of the experiment but not among surviving pigs. The present study was designed to identify and characterize the early maladaptive hemodynamic responses (tachycardia in the presence of hypotension without a corresponding increase in cardiac index or mean arterial blood pressure) and their predictive power for early mortality in this experimental model. Methods: Controlled hemorrhagic shock was induced in 16 pigs. Hemodynamic parameters were monitored continuously for 7 h following bleeding. Changes in cardiovascular and laboratory parameters were analyzed and compared between those that had arrhythmia and those that did not. Results: All animals had similar changes in parameters until the end of the bleeding phase. Six animals developed arrhythmias and died early, while 10 had no arrhythmias and survived longer than 6 h or until euthanasia. Unlike survivors, those that died did not compensate for cardiac output (CO), diastolic blood pressure (DBP), and stroke volume (SV). Oxygen delivery (DO2) and mixed venous saturation (SvO2) remained low in animals that had arrhythmia, while achieving certain measures of recuperation in animals that did not. Serum lactate increased earlier and continued to rise in all animals that developed arrhythmias. No significant differences in hemoglobin concentrations were observed between groups. Conclusions: Despite similar initial changes in variables, we found that low CO, DBP, SV, DO2, SvO2, and high lactate are predictive of death in this animal model. The results of this experimental study suggest that maladaptive responses across a range of cardiovascular parameters that begin early after hemorrhage may be predictive of impending death, particularly in situations where early resuscitative treatment may be delayed. Address reprint requests to Dr Arik Eisenkraft, MD, MHA, Institute for Research in Military Medicine, Faculty of Medicine of the Hebrew University of Jerusalem, POB 12272, Jerusalem 91120, Israel. E-mail: aizenkra@gmail.com Received 16 December, 2018 Revised 17 January, 2019 Accepted 19 June, 2019 RS and LG have equal contributions. RS contributed to all aspects of the study including study design, data acquisition, analysis, interpretation of data, and drafting/critical revision. LG and AE contributed to all aspects of the study except data acquisition and analysis respectively. LW-A and GY contributed to study design, data acquisition, and drafting/critical revision. SDG and CW contributed to interpretation of the data and drafting/critical revision. JM, GS, BS-P, and AB contributed to the study design and data acquisition. This study was supported in part by the IDF Medical Corps Grant numbers 4440520303 and 4440622654 and The Alexander Grass Foundation Fund for Research in Military Medicine, The Institute for Research in Military Medicine (IRMM), Faculty of Medicine, The Hebrew University of Jerusalem, Israel. Prof SDG is the Director of the Military Track of Medicine and The Institute for Research in Military Medicine, and the Brandman Foundation Professor of Cardiac and Pulmonary Diseases of The Faculty of Medicine, The Hebrew University of Jerusalem. Part of this work was presented in the Euroanaesthesia 2016 Congress, May 28–30, London, UK, by Dr RS. This work was presented to the Scientific Council of The Israel Medical Association as part of the research component of Dr RS' residency training in anesthesiology. The authors report no conflicts of interest. © 2019 by the Shock Society

Effects of The Poly(ADP-ribose) Polymerase Inhibitor Olaparib in Cerulein-Induced Pancreatitis Objective: Activation of the constitutive nuclear and mitochondrial enzyme poly(ADP-ribose) polymerase (PARP) has been implicated in the pathogenesis of cell dysfunction, inflammation and organ failure in various forms of critical illness. The objective of our study was to evaluate the efficacy and safety of the clinically approved PARP inhibitor olaparib in an experimental model of pancreatitis in vivo and in a pancreatic cell line subjected to oxidative stress in vitro. The preclinical studies were complemented with analysis of clinical samples to detect PARP activation in pancreatitis. Methods: Mice were subjected to cerulein-induced pancreatitis; circulating mediators and circulating organ injury markers; pancreatic myeloperoxidase and malondialdehyde levels were measured and histology of the pancreas was assessed. In human pancreatic duct epithelial cells (HPDE) subjected to oxidative stress, PARP activation was measured by PAR Western blotting and cell viability and DNA integrity were quantified. In clinical samples, PARP activation was assessed by PAR (the enzymatic product of PARP) immunohistochemistry. Results: In male mice subjected to pancreatitis, olaparib (3 mg/kg i.p.) improved pancreatic function: it reduced pancreatic myeloperoxidase and malondialdehyde levels, attenuated the plasma amylase levels, and improved the histological picture of the pancreas. It also attenuated the plasma levels of pro-inflammatory mediators (TNF-α, IL-1β, IL-2, IL-4, IL-6, IL-12, IP-10, KC) but not MCP-1, RANTES or the anti-inflammatory cytokine IL-10. Finally, it prevented the slight, but significant increase in plasma blood urea nitrogen level, suggesting improved renal function. The protective effect of olaparib was also confirmed in female mice. In HPDE cells subjected to oxidative stress olaparib (1 μM) inhibited PARP activity, protected against the loss of cell viability and prevented the loss of cellular NAD+ levels. Olaparib, at 1–30 μM did not have any adverse effects on DNA integrity. In human pancreatic samples from patients who died of pancreatitis, increased accumulation of PAR was demonstrated. Conclusion: Olaparib improves organ function and tempers the hyperinflammatory response in pancreatitis. It also protects against pancreatic cell injury in vitro without adversely affecting DNA integrity. Repurposing and eventual clinical introduction of this clinically approved PARP inhibitor may be warranted for the experimental therapy of pancreatitis. Address reprint requests to Csaba Szabo, MD, PhD, Chair of Pharmacology, Faculty of Science and Medicine, University of Fribourg, Chemin du Musée 18, Fribourg 1700, Switzerland. E-mail: csaba.szabo@unifr.ch. Received 22 May, 2019 Revised 18 June, 2019 Accepted 26 June, 2019 Ethics approval and consent to participate: The current report does not contain human interventional studies. (The human pancreatic tissue samples were obtained from a commercial biobank which obtained them under informed consent and in compliance with all applicable rules and regulations.) Consent for publication: The current report does not contain human studies or studies that would require consenting. Availability of data and material: The datasets during and/or analyzed during the current study available from the corresponding author on reasonable request. Competing interests: The authors declare no competing interests. Funding: This work was supported by grants from the National Institutes of Health (R01GM107876) and the Swiss National Foundation (to C.S.), and by the FAPESP (to R.S. and F.G.S.) Authors' contributions: AA conducted in vivo and in vitro experiments; BS, GT, ND, and MM conducted in vitro studies, CS, LL, FGS and RS conceived the overall study design; AA, ST, LL, FGS and RS contributed to various parts of the study design, method development, data interpretation and writing of the manuscript. CS was responsible for the coordination of the project. CS drafted the first version of the manuscript and finalized the manuscript. All authors read and approved the final manuscript. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). The authors declare that they have no competing interests. © 2019 by the Shock Society

Activation of hypoxia-inducible factor -1α via succinate dehydrogenase pathway during acute lung injury induced by trauma/hemorrhagic shock Hypoxia-inducible factor (HIF)-1α is a transcription factor that is critical for tissue adaption to hypoxia and inflammation. Previous studies had indicated that normoxic activation of HIF-1α in cancer involves inhibition or mutation of the metabolic enzyme succinate dehydrogenase (SDH). We have found that local inhibition of HIF-1α ameliorates acute lung injury (ALI) induced by trauma/hemorrhagic shock (T/HS) in rats. In this study, we found pulmonary activation of HIF-1α and inhibition of SDH during THS-Induced ALI in rats and transcriptional activation of HIF-1α during ALI induced by T/HS lymph via SDH pathway in vitro. Furthermore, pharmacologic inhibition of HIF-1α attenuates lung inflammation and pulmonary edema during ALI by T/HS. Activation of HIF-1α is detrimental to ALI induced by T/HS. Thus, our data suggest that HIF-1α activation by T/HS is necessary for T/HS-induced lung injury and a critical role for SDH in the initiation of acute inflammatory response following ALI. Nevertheless, this is a preclinical work and several limitations impede translation of the findings to patients, such as uncontrolled bleeding and simultaneous treatment, and prolonged course of clinical shock on the outcome of the work, which needs to be addressed in future. Address reprint requests to Qifang Li, Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, NO.2 Ruijin Road, Shanghai, 200025, China. E-mail: psoriasisbreak@163.com. Received 25 December, 2018 Revised 23 January, 2019 Accepted 14 March, 2019 There is no relevant conflict of interest. © 2019 by the Shock Society

Identification of Fibrinogen as a Key Anti-Apoptotic Factor in Human Fresh Frozen Plasma for Protecting Endothelial Cells In Vitro Resuscitation with human fresh frozen plasma (FFP) in hemorrhagic shock (HS) patients is associated with improved clinical outcomes. Our group has demonstrated that the beneficial effect of FFP is due to its blockade on endothelial hyperpermeability, thereby improving vascular barrier function. The current study aimed to investigate HS-induced endothelial cell apoptosis, a potential major contributor to the endothelial hyperpermeability, and to determine the effect and the key components/factors of FFP on protecting endothelial cells from apoptosis. We first measured and demonstrated an increase in apoptotic endothelial microparticles (CD146+AnnexinV+) in patients in shock compared to normal subjects, indicating the induction of endothelial cell activation and apoptosis in shock patients. We then transfused HS rats with FFP and showed that FFP blocked HS-induced endothelial cell apoptosis in gut tissue. To identify the anti-apoptotic factors in FFP, we utilized high-performance liquid chromatography, fractionated FFP, and screened the fractions in vitro for the anti-apoptotic effects. We selected the most effective fractions, performed mass spectrometry, and identified fibrinogen as a potent anti-apoptotic factor. Taken together, our findings suggest that HS-induced endothelial apoptosis may constitute a major mechanism underlying the vascular hyperpermeability. Furthermore, the identified anti-apoptotic factor fibrinogen may contribute to the beneficial effects of FFP resuscitation, and therefore, may have therapeutic potential for HS. Address reprint requests to Yanna Cao, MD, Department of Surgery, The University of Texas Health Science Center at Houston, 6431 Fannin St., MSB4.608, Houston, TX 77030. E-mail: Yanna.Cao@uth.tmc.edu, Tien C. Ko, MD, Department of Surgery, The University of Texas Health Science Center at Houston, 5656 Kelly, 30S62008, Houston, TX 77026. E-mail: Tien.C.Ko@uth.tmc.edu. Received 9 May, 2019 Revised 28 May, 2019 Accepted 14 June, 2019 This study was supported by the National Institute of General Medical Sciences P50 grant GM038529 (T.C.K. and J.B.H.), Jack H Mayfield M.D. Distinguished Professorship in Surgery (T.C.K), the William Stamps Farish Fund, the Howell Family Foundation, and the James H. "Red" Duke Professorship Chair fund (C.E.W), and Dean's fund for Summer Research Program (J.M.D.). The authors report no conflicts of interest. © 2019 by the Shock Society

First-Days Reduction of Plasma and Skin Advanced Glycation End Products is Related to Outcome in Septic Patients Background: Advanced glycation end products (AGEs) are a result of non-enzymatic glycation of proteins and lipids, which can attach to either their cell surface receptor (RAGE) or its soluble form (sRAGE). Evidence exists for the implication of AGE-RAGE axis in sepsis, but data are still insufficient and conflicting. We aimed to analyse the kinetics of plasma and skin AGEs and sRAGE during sepsis, and their association with outcome in septic patients. Methods: We performed a prospective observational study. We enrolled 90 consecutive patients with severe sepsis or septic shock, within the first 24 hours of Intensive Care Unit admission. During the first 5 days of sepsis, we measured plasma autofluorescence (PAF) and skin autofluorescence (SAF) as surrogates of circulating and skin AGEs, respectively. sRAGE was measured on days 1, 3 and 5. Delta values were defined as the difference between the PAF, SAF or sRAGE on a specific day and the value on day 1. Results: 28-day mortality was 18%. Bivariate analysis found that ΔPAF3-1, ΔPAF4-1, ΔPAF5-1 and ΔSAF5-1 were significantly associated with 28-day mortality. Additionally, sRAGE1 was inversely correlated to ΔPAF4-1 (r = -0.250, p = 0.019) and ΔPAF5-1 (r = -0.246, p = 0.024), and significantly associated with 28-day mortality. In an adjusted multivariate logistic regression analysis, ΔPAF2-1, ΔPAF3-1, ΔPAF4-1, ΔPAF5-1 and ΔSAF5-1 were associated with 28-day mortality. Conclusions: Kinetics of plasma and skin AGEs during the first days of sepsis are independently associated with mortality, where a decrease of plasma and skin AGEs are related to higher mortality. Address reprint requests to Emilio Rodriguez-Ruiz, MD, Intensive Care Medicine Department, Complexo Hospitalario Universitario de Santiago (CHUS). C/Choupana s/n, 15706 Santiago de Compostela, A Coruña, Spain. E-mail: r.ruizemilio@gmail.com Received 27 March, 2019 Revised 10 June, 2019 Accepted 10 June, 2019 Conflicts of interest and Source of Funding: The authors have no conflicts of interest to declare regarding this article. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

Myocardial Edema: An Overlooked Mechanism of Septic Cardiomyopathy? No abstract available

Catecholamines Induce Endoplasmic Reticulum Stress via Both Alpha and Beta Receptors Severely burned patients suffer from a hypermetabolic syndrome that can last for years after the injury has resolved. The underlying cause of these metabolic alterations most likely involve the persistent elevated catecholamine levels that follow the surge induced by thermal injury. At the cellular level, endoplasmic reticulum (ER) stress in metabolic tissues is a hallmark observed in patients following burn injury and is associated with several detrimental effects. Therefore, ER stress could be the underlying cellular mechanism of persistent hypermetabolism in burned patients. Here, we show that catecholamines induce ER stress and that adreno-receptor blockers reduce stress responses in the HepG2 hepatocyte cell line. Our results also indicate that norepinephrine (NE) significantly induces ER stress in HepG2 cells and 3T3L1 mouse adipocytes. Furthermore, we demonstrate that the alpha-1 blocker, prazosin, and beta blocker, propranolol, block ER stress induced by NE. We also show that the effects of catecholamines in inducing ER stress are cell type-specific, as NE treatment failed to evoke ER stress in human fibroblasts. Thus, these findings reveal the mechanisms used by catecholamines to alter metabolism and suggest inhibition of the receptors utilized by these agents should be further explored as a potential target for the treatment of ER stress-mediated disease. Address reprint requests to Marc G. Jeschke, MD, PhD, Director Ross Tilley Burn Centre, Sunnybrook Health Sciences Centre; Division of Plastic Surgery, Department of Surgery, Department of Immunology, University of Toronto; Sunnybrook Research Institute, 2075 Bayview Ave., Rm D704, Toronto, ON, CANADA, M4N 3M5. E-mail: marc.jeschke@sunnybrook.ca Received 18 March, 2019 Revised 5 April, 2019 Accepted 3 June, 2019 Disclosure statement: The authors have nothing to disclose. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

TGF-β Pathway Inhibition Protectsthe Diaphragm From Sepsis-Induced Wasting and Weakness in Rat Sepsis is a frequent complication in patients in intensive care units (ICU). Diaphragm weakness,one of the most common symptoms observed, can lead to weaning problemsduring mechanical ventilation. Over the last couple of years, members of thetransforming growth factor (TGF) β family,such as myostatin, activin A and TGF-β1,have beenreported to strongly trigger the activation of protein breakdown involved in muscle wasting. The aim of this study was to investigate the effect of TGF-βinhibitorLY364947on the diaphragm during chronic sepsis. Rats were separated into four groups exposed to different experimental conditions: (i) Control group, (ii) Septic group, (iii) Septic group with inhibitor from day 0 (LY D0), and (iv) Septic group with inhibitor from day 1 (LY D1). Sepsis was induced in ratsby cecal ligation and puncture, and carried out for seven days. Chronic sepsis was responsible for a decrease inbody weight, food intake anddiaphragm's mass. The inhibitor was able to abolish diaphragm wasting only in the LY D1 group. Similarly, LY364947 had a beneficial effect on the diaphragm contraction only for the LY D1 group. SMAD3 was over-expressed and phosphorylated within rats in the Septic group;however this effect was reversed by LY364947. Calpain-1 and -2 as well as MAFbx were over-expressedwithin individuals in theSeptic group. Yet, calpain-1 and MAFbx expressions were decreased by LY364947 With this work, we demonstrate for the first time that the inhibition of TGF-β pathway during chronic sepsisprotects the diaphragm from wasting and weaknessas early asone day post infection. This could lead to more efficient treatment and care for septic patients in ICU. Address reprint requests to Jude Baptiste, PhD, Laboratoire de Physiologie – EA4324 ORPHY, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins 29238 Brest, Cedex 3 France; E-mail: bjude@outlook.fr Received 14 September, 2018 Revised 15 October, 2018 Accepted 29 May, 2019 Highlights: -Chronic sepsis of seven days is responsible for the decrease inthe diaphragm mass and its contractile force. -Sepsis induces the activation of TGF-β signaling and proteolytic pathway in the diaphragm. -The inhibition of TGF-β pathway prevents diaphragm dysfunction only if the inhibition starts 24 h after sepsis induction. -The early inhibition of the TGF-β pathway does not protect the diaphragm during sepsis. Conflict of interest: The authors declare that no conflict of interest exists. © 2019 by the Shock Society

The Use of Tranexamic acid (TXA) for The Management of Haemorrhage In Trauma Patients In The Prehospital Environment: Literature Review and Descriptive Analysis of Principal Themes Tranexamic acid (TXA) is an anti-fibrinolytic agent used to prevent traumatic exsanguination. It was first introduced to clinical practice for the management of patients with bleeding disorders, especially adapted to reduce bleeding in haemophiliacs undergoing oral surgical interventions. TXA exerts its action on the coagulation process by competitively inhibiting plasminogen activation, thereby reducing conversion of plasminogen into plasmin. This ultimately prevents fibrinolysis and reduces haemorrhage. Thus, TXA may be well suited for the management of traumatic haemorrhage in the pre-hospital setting. Despite multiplicity of studies on the use of TXA in clinical practice, there is no consensus regarding the use of TXA for the management of haemorrhage in trauma patients in the prehospital environment. Thus, a review on this topic was warranted. An extensive literature search yielded 14 full journal articles which met the inclusion criteria. These articles were thoroughly analysed and the and following themes were identified: "Dose of TXA administration", "Route of TXA administration", "Optimal window of TXA administration", "Safety of TXA use", "Clinical Effectiveness of TXA application" and the "Feasibility of TXA use in the prehospital setting". Overall, to achieve the best possible outcomes, literature supports the use of a loading dose of 1 gram of TXA, followed by 1 gram infusion over 8 hours, given by intravenous administration within a 3 hour window period of traumatic injury. TXA is very effective and safe to use in the pre-hospital setting, and its use is clinically and economically feasible. Address reprint requests to Dr Emmanuel Jesulola, Room 206, Building 1448, Charles Sturt University, Panorama Avenue, Bathurst, NSW 2795 Australia. E-mail: ejesulola@csu.edu.au Received 4 April, 2019 Revised 18 April, 2019 Accepted 14 May, 2019 Funding: No funding was required for this review article. Disclosure of interest: The authors report no conflict of interest. © 2019 by the Shock Society

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

A&A Practice - anesthesia-analgesia

Decreasing Frontal Electroencephalogram Alpha Power and Increasing Sensitivity to Volatile Anesthetics Over 3 Surgeries Within 7 Months: A Case Report Depth of anesthesia (DoA) monitors are widely used during general anesthesia to guide individualized dosing of hypnotics. Other than age and specific drugs, there are few reports on which comorbidities may influence the brain and the resultant electroencephalogram (EEG) of patients undergoing general anesthesia. We present a case of a patient undergoing 3 cardiac operations within 7 months with severe illness and comorbidity, leading to pronounced physical frailty and significant changes of frontal alpha power in the EEG and increased sensitivity to volatile anesthetics. These findings may have important clinical implications and should trigger further investigations on this topic. Accepted for publication May 1, 2019. Funding: This work received funding from Bangerter-Rhyner Foundation Grant and Clinical Trials Unit grant of the University Hospital of Bern, Inselspital, Bern, Switzerland. The authors declare no conflicts of interest. Clinical trial number: ClinicalTrials.gov; Identifier: NCT02976584. Address correspondence to, Heiko A. Kaiser, MD, Department of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, Freiburgstrasse, 3010 Bern, Switzerland., Address e-mail to heiko.kaiser@insel.ch. © 2019 International Anesthesia Research Society

Postpartum Thyroid Storm in Poorly Controlled Graves' Disease: A Case Report Hyperthyroidism during pregnancy is a relatively rare event, but poor control of hyperthyroidism during pregnancy is associated with a host of issues. These include intrauterine fetal demise, hypertensive disorders of pregnancy, preterm delivery, low birth weight, intrauterine growth restriction, and maternal congestive heart failure. One of the most feared sequelae of hyperthyroidism is thyroid storm, which has a mortality rate >10%. Patients who develop thyroid storm present several challenges to anesthesiologists including hemodynamic instability and medication management. Here, we present the anesthetic management for a parturient whose delivery was complicated by signs and symptoms consistent with thyroid storm. Accepted for publication May 23, 2019. Funding: None. The authors declare no conflicts of interest. Address correspondence to Jack M. Peace, MD, Department of Anesthesiology, Northwestern University Feinberg School of Medicine, 251 E Huron F5-704, Chicago, IL 60611. Address e-mail to jack.peace@northwestern.edu. © 2019 International Anesthesia Research Society

Analgesic Rescue With Opioid-Only Thoracic Epidural After Surgical Infiltration of Liposomal Bupivacaine: A Case Report We present the case of a 51-year-old man with a history of recurrent lung cancer after left upper lobectomy who presented for an elective completion pneumonectomy via a bilateral anterior thoracotomy incision. At the completion of surgery, bilateral multilevel intercostal infiltration was performed with liposomal bupivacaine. Due to poorly controlled postoperative pain after extubation, a thoracic epidural was placed in the intensive care unit. An opioid-only infusion was started and transitioned to a local anesthetic–based infusion on postoperative day 2. This case report represents a novel stepwise approach of thoracic epidural management after surgical infiltration of liposomal bupivacaine. Accepted for publication May 16, 2019. Funding: None. The authors declare no conflicts of interest. Address correspondence to Matthew W. Swisher, MD, Department of Anesthesiology, University of California San Diego, 9500 Gilman Dr MC 0898, La Jolla, CA 92093. Address e-mail to mwswisher@ucsd.edu. © 2019 International Anesthesia Research Society

Lung Isolation for Whole Lung Lavage in a Pediatric Patient With Atypical Airway Anatomy due to Short Stature: A Case Report This case demonstrates the airway management of a pediatric patient with short stature due to STAT5b deficiency, a rare genetic immunodeficiency associated with lung disease and endocrinopathy. The patient had recurrent pulmonary infections and pulmonary alveolar proteinosis (PAP) for which whole lung lavage (WLL) was recommended. Due to short stature and overall body habitus, the patient's airway would not accommodate a traditional double-lumen tube (DLT). Therefore, we placed 2 single-lumen breathing tubes: 1 endobronchial and 1 endotracheal, to mimic a DLT and facilitate WLL, demonstrating a viable option for lung isolation in the absence of purpose-built equipment. Accepted for publication April 30, 2019. Funding: None. The authors declare no conflicts of interest. Address correspondence to Susan E. Eklund, MD, Department of Anesthesiology, Critical Care and Pain Medicine, Boston Children's Hospital, 300 Longwood Ave, Bader 3, Boston, MA 02115. Address e-mail to susan.eklund@childrens.harvard.edu. © 2019 International Anesthesia Research Society

Comparison of Twitter Use of the American Society of Anesthesiologists and the American Association of Nurse Anesthetists Twitter has become a powerful tool for dissemination of information. The objective of this study was to evaluate Twitter usage of the American Society of Anesthesiologists (ASA) and the American Association of Nurse Anesthetists (AANA). All tweets from ASA ("@ASALifeline") and AANA ("@aanawebupdates") were collected over a 1-year time period. The content of each tweet was categorized using a rubric. ASA generated more original tweets than AANA. Twitter use was highest in October for ASA and September for AANA. Both societies are actively using Twitter. Future work should evaluate the impact of societal Twitter use. Accepted for publication May 7, 2019. Funding: None. The authors declare no conflicts of interest. Address correspondence to Elizabeth M. S. Lange, MD, Department of Anesthesiology, Northwestern University Feinberg School of Medicine, 251 E Huron St, F5-704, Chicago, IL 60611. Address e-mail to elizabeth.lange@northwestern.edu. © 2019 International Anesthesia Research Society

Having a Conscious Patient During Cardiopulmonary Resuscitation: Is It Not Time to Consider Sedation Protocol?: A Case Report A middle-aged man with acute inferior wall myocardial infarction was admitted in cardiac arrest and in an unresponsive state to the hospital. Cardiopulmonary resuscitation (CPR) was initiated. Patient showed signs of consciousness throughout the CPR. The impact of awareness during CPR on the neuropsychological status of a patient with a favorable neurological outcome is yet to be studied on a large scale. Sedation protocol without compromising hemodynamic status may prove a fair choice in such cases. Accepted for publication April 9, 2019. Funding: None. The author declares no conflicts of interest. Address correspondence to Vasanth Sukumar, DA, DNB, Royal Care Super Specialty Hospitals, Neelambur, Coimbatore 641062, Tamil Nadu, India. Address e-mail to vasnt.dr@gmail.com. © 2019 International Anesthesia Research Society

Bilateral Sciatic Neuropathies as a Complication of Positioning During Neuraxial Anesthesia for Cesarean Delivery: A Case Report Neurologic complications following neuraxial anesthesia for cesarean delivery are rare. We present a 33-year-old parturient who developed prolonged lower extremity weakness following a single-shot subarachnoid block for cesarean delivery. After neurologic evaluation, she was diagnosed with bilateral sciatic neuropathies due to prolonged positioning for the anesthetic. We review the incidence of nerve injury associated with neuraxial anesthesia and risk factors for developing peripheral nerve injury in this context. We offer a solution to prevent this complication from occurring. Accepted for publication March 18, 2019. Funding: None. The authors declare no conflicts of interest. The views expressed are those of the authors and do not reflect the official views or policy of the Department of Defense or its components. Address correspondence to Bradley A. Reel, MD, Department of Anesthesia, Brooke Army Medical Center, 3551 Roger Brooke Dr, San Antonio, TX 78234. Address e-mail to bradley.a.reel2.mil@mail.mil. © 2019 International Anesthesia Research Society

Andexanet Alfa for Urgent Reversal of Apixaban Before Aortic Surgery Requiring Cardiopulmonary Bypass: A Case Report Andexanet alfa is a recombinant factor Xa decoy molecule capable of reversing direct and indirect factor Xa–inhibiting anticoagulants. We present an adult patient on apixaban for nonvalvular atrial fibrillation who required urgent reoperative aortic surgery for an aortic root pseudoaneurysm. Apixaban was reversed with andexanet alfa. A second dose of andexanet alfa was required before surgical incision for persistently elevated antifactor Xa levels. Intraoperative management required use of cardiopulmonary bypass (CPB). No major adverse cardiovascular, cerebrovascular, hemorrhagic, or thromboembolic events were observed. Accepted for publication May 13, 2019. The authors declare no conflicts of interest. Funding: None. Address correspondence to Devon Flaherty, MD, Department of Anesthesiology, Perioperative, and Pain Medicine, Brigham and Women's Hospital, 75 Francis St, Boston, MA 02115. Address e-mail to flaherdc@gmail.com. © 2019 International Anesthesia Research Society

Large Iatrogenic Tracheal Injury From Attempted Endotracheal Cuff Leak Remedy: A Case Report Continuous insufflation of oxygen into the pilot balloon of an endotracheal tube has been documented as a viable method to fix an incompetent endotracheal tube cuff. We present a case of a large iatrogenic tracheal injury resulting from an improvised cuff leak management device. Valuable lessons are discussed to prevent similar incidents. Accepted for publication April 10, 2019. Funding: None. The authors declare no conflicts of interest. Supplemental digital content is available for this article. Direct URL citations are provided in the HTML and PDF versions of this article on the journal's website (www.cases-anesthesia-analgesia.org). Address correspondence to Ron E. Samet, MD, Division of Trauma Anesthesiology, University of Maryland Medical Center, 22 S Greene St, Room T1R77, Baltimore, MD 21201. Address e-mail to rsamet@som.umaryland.edu. © 2019 International Anesthesia Research Society

Electroencephalogram Suppression Despite Extremely Low–Dose Anesthetic During Cardiac Surgery: A Case Report A prominent feature of electroencephalogram (EEG) under general anesthesia is anteriorization of α waves. We report the case of a 63-year-old man anesthetized for coronary artery bypass grafting in whom the NeuroSENSE-processed EEG monitor recorded only δ waves in the absence of α frequencies, along with high EEG suppression despite extremely low doses of anesthetics during the whole procedure. The patient fully recovered from anesthesia 2 hours after the procedure and showed neither awareness nor neurological complications. This atypical EEG pattern under low concentration of anesthetics may be an intraoperative marker of a specific brain phenotype. Accepted for publication April 15, 2019. Funding: None. Conflicts of Interest: See Disclosures at the end of the article. Address correspondence to Victoria Van Regemorter, MD, Anesthesiology, Cliniques Universitaires St-Luc, Université Catholique de Louvain, Ave Hippocrate 10, 1200 Bruxelles, Belgium. Address e-mail to victoria.vanregemorter@uclouvain.be. © 2019 International Anesthesia Research Society

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Conservation Genetics

Population management using gene drive: molecular design, models of spread dynamics and assessment of ecological risks Abstract CRISPR gene drive has recently been proposed as a promising technology for population management, including in conservation genetics. The technique would consist in releasing genetically engineered individuals that are designed to rapidly propagate a desired mutation or transgene into wild populations. Potential applications in conservation biology include the control of invasive pest populations that threaten biodiversity (eradication and suppression drives), or the introduction of beneficial mutations in endangered populations (rescue drives). The propagation of a gene drive is affected by different factors that depend on the drive construct (e.g. its fitness effect and timing of expression) or on the target species (e.g. its mating system and population structure). We review potential applications of the different types of gene drives for conservation. We examine the challenges posed by the evolution of resistance to gene drives and review the various molecular and environmental risks associated with gene drives (e.g. propagation to non target populations or species and unintended detrimental ecosystem impacts). We provide some guidelines for future gene drive research and discuss ethical, biosafety and regulation issues.

Conservation implications of widespread polyploidy and apomixis: a case study in the genus Pomaderris (Rhamnaceae) Abstract Polyploidy resulting from whole genome duplication is common in plants and is increasingly being recognised as a critical factor in conservation actions, particularly when within-species variation in ploidy exists. Pomaderris(Rhamnaceae), a genus of 70 species endemic to Australia and New Zealand, has many species listed as threatened and that are subject to conservation management but of unknown ploidy. To provide a better understanding of polyploidy in the genus we sampled 36 of 70 species of Pomaderris and used flow cytometry to establish genome sizes and infer ploidy. Additionally, to examine within-species variation, we screened 104 individuals of eight rare species subject to conservation management. We did not find evidence for infraspecific variation in ploidy, suggesting that from a cytological perspective, conspecific individuals from the screened populations do not need to be kept geographically separated in conservation management. There is, however, considerable variation among species, with genome sizes suggesting the occurrence of diploidy, triploidy, tetraploidy and hexaploidy. Finding several species to be triploid but capable of seed production, we then explored reproductive biology using the flow cytometric seed screen. Results suggested that triploid species produce seeds asexually, as previously reported for two New Zealand congeners. While asexual reproduction through apomixis is generally a means of odd-ploid taxa overcoming sterility, we found that more than half of examined diploids and tetraploids also produced seeds asexually. Asexual reproduction means genetic diversity is potentially low, and these results should therefore be considered in future conservation actions and seed sampling designs.

Comparative landscape genetics of two endemic torrent salamander species, Rhyacotriton kezeri and R. variegatus : implications for forest management and species conservation Abstract Comparative landscape genetic studies provide insights into whether relationships between landscape features and patterns of spatial genetic structure differ among populations, species, habitat types, and regions. For species with fragmented distributions, especially when management practices contribute to fragmentation, tests of the factors structuring population connectivity are particularly important for understanding continued risks. We determined levels of genetic diversity and tested the relationships of landscape-scale vegetative, geographic, and climate variables with genetic distance in two congeneric, endemic salamander species with status of concern. Using microsatellite data for 326 Rhyacotriton kezeri and 557 Rhyacotriton variegatus individuals collected from 17 to 29 localities, respectively, we implemented a model of landscape resistance based on circuit theory. The northernmost portions of each species' range is more fragmented than areas to the south, leading to the prediction that these areas would have relatively lower genetic diversity in response. Due to reliance of both species upon cold-water habitats, we predicted that landscape variables maintaining cool, moist microhabitats would be correlated with gene flow. Genetic structure was high overall and trended toward increasing with the proportion of the forested landscape. Based on maximum likelihood population effects models across genetic clusters and species, land cover and roads were the best predictors of genetic distance, even though the degree of fragmentation differed across each species' geographic range. Our results suggest that forest cover is essential for dispersal in these salamanders, indicating negative effects of fragmentation resulting from timber harvest and other forest disturbances.

Impacts of stocking and its intensity on effective population size in Brook Charr ( Salvelinus fontinalis ) populations Abstract Effective population size (Ne) is a measure of the genetic size of a population and a crucial parameter for wildlife population management since it is strongly related to retention of genetic diversity in time and/or to inbreeding levels. Many exploited fish populations are stocked with the purpose of increasing population sizes to sustain important fishing pressures. However, stocking hatchery-reared fish could at the same time increase population census size and decrease Ne. Our study aimed at characterizing how stocking affected Ne in supplemented populations of Brook Charr (Salvelinus fontinalis) in Québec and at assessing how this relationship varied with the intensity of stocking (e.g. number of stocking events, number of fish stocked/ha, proportion of domestic genetic background). We estimated Ne with the linkage disequilibrium method in 54 populations (3361 sampled individuals analyzed at 20 microsatellites) with various levels of stocking intensity. We found that stocked lakes have significantly lower Ne than unstocked lakes. However, we found little evidence of an additional effect of stocking intensity on Ne of stocked lakes. Our results suggest that stocking may have a negative impact on Ne but that more intense stocking does not necessarily translate into lower Ne. However, even though low Ne in stocked populations could be attributed to an effect of stocking, it is also likely that stocked lakes consist of poor environments that translate into low Ne.

Predicting population extinctions in Darwin's finches Abstract Genetic data are increasingly used for fast, efficient, and cost-effective monitoring of natural populations and assessment of extinction risk in species management. A single modern molecular snapshot is typically used to infer population size and vulnerability, yet for species with unknown and potentially complex genetic metapopulation structure, this technique may not effectively predict vulnerability. Darwin's finches, which are well-represented in museum collections, offer a unique opportunity to test the effectiveness of predicting extinction vulnerability in species with complex structure, such as naturally fragmented populations. In this study, we compared ancient DNA from ~ 100 year old extinct and extant Darwin's finch populations in the Galápagos Islands to determine whether single time point genetic assessments in the past accurately predicted extinction risk, or if other factors such as metapopulation dynamics could mask population declines. Of eight extinct populations, only one had significantly reduced genetic variation compared to an extant population of similar characteristics. Contrary to our prediction that populations would have decreased genetic diversity prior to extinction when compared to persisting populations, at least one measure of genetic diversity was significantly higher in six of the eight extinct populations when compared to extant populations. Simulations lend support to the hypothesis that unaccounted for metapopulation structure may explain the observed pattern in many species. Therefore, models of genetic diversity reflecting population extinction potential may be inadequate for highly-mobile species with metapopulation dynamics such as the Galápagos finches.

Genetic structure of an abundant small mammal is influenced by low intensity urbanization Abstract Urban development can fragment and degrade habitat, and such habitat alterations can have profound effects on wildlife, including influencing population genetics. We used nine microsatellite loci to determine the effects of urbanization on genetic diversity and genetic structure in a native small mammal, Merriam's kangaroo rat, in areas in and around Las Cruces, NM, an expanding low density urban center. We found that Merriam's kangaroo rats in urban areas had increased genetic differentiation among populations as compared to wildland animals, and detected some evidence of lower genetic diversity in urban areas, indicating that the population genetics of a common and abundant wildlife species can be impacted by low density urbanization. Our results suggest that although abundant and common wildlife such as Merriam's kangaroo rats may persist in urban environments, these animals may still be influenced by more subtle effects of urbanization, such as genetic isolation.

Genetic mixture analyses in support of restoration of a high value recreational fishery for rainbow trout ( Oncorhynchus mykiss ) from a large lake in interior British Columbia Abstract Genetic mixture analysis is an important tool to apportion catch amongst potential component populations contributing to a fishery. We used variation at 10 microsatellite DNA loci to assess the level of genetic divergence between two ecotypes of rainbow trout (Oncorhynchus mykiss) that naturally co-occur in Kootenay Lake, southeastern British Columbia, and to exploit such divergence in a mixture analysis. One form, "Gerrard" rainbow trout, historically matured at sizes greater than 60 cm and 5 kg, spawns at a lake outlet after upstream migration in a large river tributary to Kootenay Lake, and is highly prized in the recreational fishery. The other form, "non-Gerrard" rainbow trout, is also native to the lake and matures at smaller sizes and spawns in numerous small streams tributary to Kootenay Lake. Recent declines in growth rate of Gerrard rainbow trout, however, has made them difficult to identify by size in fishery samples. Gerrard (N = 130, 6 sites) and non-Gerrard trout N = 312, 15 sites) were highly divergent from one another (FST = 0.14, P < 0.001) and constituted distinct genetic groups in model-based clustering analyses. Genetic mixture analyses of fishery samples indicated a high degree of accuracy in estimating mixture proportions; 100% Gerrard simulated fisheries were estimated to contain 99.9% Gerrards (95% confidence intervals of 99.8–100%) while 100% non-Gerrard rainbow trout simulated fisheries were estimated to contain 100.0% non-Gerrard trout (100–100%). Across eight fishery creel samples obtained between 2015 and 2017 (N = 527 fish), mixture analysis estimated the fishery to contain an average of 73.4% (95% confidence interval = 68.4–74.6%) Gerrard and 26.6% (23.4–31.6%) non-Gerrard trout. Realistic fishery simulations demonstrated strong agreement with empirical results; the average simulated values for Gerrards was 73.4% (65.9–80.0%) and for non-Gerrards was 26.8% (20.0–34.1%). Assignment tests resulted in an average 98.5% (± 0.066%) assignment confidence; 385 fish from the fishery samples were assigned to the Gerrard group (0.73) and 142 (0.27) to the non-Gerrard rainbow trout genetic group. Fitting length-at-age data for genetically assigned fishery samples to a von Bertalanffy growth model found greatest support for a model employing ecotype-specific L∞ (= 59.6 and 52.9 cm for Gerrards and non-Gerrards, respectively, both P < 0.001), and t0 (= − 1.36 and − 2.58, respectively, both P < 0.05), but a common K (= 0.189, P < 0.001). Our mixture analyses are being used to monitor catches and better understand the feeding and migration biology of these sympatric ecotypes of rainbow trout.

Messing about on the river: the role of geographic barriers in shaping the genetic structure of Bornean small mammals in a fragmented landscape Abstract Landscape features may restricting dispersal and gene flow, and increase demographic isolation among sub-populations. In addition, landscape features may represent potential dispersal barriers depending on species vagility. To predict the persistence of populations and to formulate adequate conservation measures it is essential to understand the ability of species to transverse landscape barriers. Using population genetic techniques we assessed the importance of physical barriers along the Kinabatangan River for a suite of non-volant small mammals. Cytochrome b sequence variation was examined for each of the 19 species sampled across both riverbanks. Haplotype networks and molecular variance analyses indicated contrasting patterns of genetic isolation between riversides for different taxa. Genetic isolation between riversides ranged from moderate to complete in tree shrews and squirrels, whereas no isolating effect could be detected in murids and gymnures. Although genetic divergence between forest fragments on the same side of the river could only be studied in a subset of six species, the results suggest an additional dispersal barrier for two of these studied species. While barrier effects of a paved road and tributaries could not be verified, large oil palm plantations seem to have disrupted gene flow in these species. Furthermore, the findings suggest higher genetic connectivity on the more continuously forested compared to the more fragmented riverside, and underline the importance of forest corridors as essential conservation measures to maintain genetic diversity in a fragmented landscape such as that along the Kinabatangan River.

History matters: contemporary versus historic population structure of bobcats in the New England region, USA Abstract Habitat fragmentation and genetic bottlenecks can have substantial impacts on the health and management of wildlife species by lowering diversity and subdividing populations. Population genetic comparisons across time periods can help elucidate temporal changes in populations and the processes responsible for the changes. Bobcats (Lynx rufus) are wide-ranging carnivores and are currently increasing in abundance across an expanding range. Bobcat populations in New England have fluctuated in the past century in response to changes in their prey base, harvest pressure, and landscape development. We genotyped contemporary (2010–2017) and historic (1952–1964) bobcats from New England and Quebec, Canada at a suite of microsatellite loci and tested for differences in diversity, effective population size, and gene flow. Over 20 generations separated the sampling periods, and the intervening years were marked by drastic changes in land use and species management regimes. We found a general decrease in genetic diversity and differing population genetic structure through time. Effective population size decreased at the end of the historic period, coincident with a spike in harvest, but rebounded to greater numbers in the contemporary period. Our results suggest that bobcat populations in the region are robust, but development and range dynamics may play a significant role in population structure. Our study also highlights the benefits of a historical perspective in interpreting contemporary population genetic data.

Beyond Bonferroni revisited: concerns over inflated false positive research findings in the fields of conservation genetics, biology, and medicine Abstract In 2006, Narum published a paper in Conservation Genetics emphasizing that Bonferroni correction for multiple testing can be highly conservative with poor statistical power (high Type II error). He pointed out that other approaches for multiple testing correction can control the false discovery rate (FDR) with a better balance of Type I and Type II errors and suggested that the approach of Benjamini and Yekutieli (BY) 2001 provides the most biologically relevant correction for evaluating the significance of population differentiation in conservation genetics. However, there are crucial differences between the original Benjamini and Yekutieli procedure and that described by Narum. After carefully reviewing both papers, we found an error due to the incorrect implementation of the BY procedure in Narum (Conserv Genet 7:783–787, 2006) such that the approach does not adequately control FDR. Since the incorrect BY approach has been increasingly used, not only in conservation genetics, but also in medicine and biology, it is important that the error is made known to the scientific community. In addition, we provide an overview of FDR approaches for multiple testing correction and encourage authors first and foremost to provide effect sizes for their results; and second, to be transparent in their descriptions of multiple testing correction. Finally, the impact of this error on conservation genetics and other fields will be study-dependent, as it is related to the number of true to false positives for each study.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Epidemiology

Extending inferences from a randomized trial to a target population

New methods for generalizability and transportability: the new norm

Coffee consumption and all-cause and cause-specific mortality: a meta-analysis by potential modifiers Abstract Coffee consumption has been associated with decreased mortality in previous studies. As aging, obesity, and lifestyle factors affect the risk of mortality, the association between coffee and mortality needs to be examined in various subpopulations by characteristics of subjects. To quantitatively assess this association, we conducted an updated meta-analysis including stratified analyses by potential modifiers. We searched in the PubMed and Web of Science databases through March 8, 2019, and conducted meta-analysis including linear and non-linear dose–response analyses. We identified 40 studies including 3,852,651 subjects and 450,256 all-cause and cause-specific deaths. Non-linear inverse associations between coffee consumption and mortality from all-causes, cardiovascular disease (CVD), and cancers were found. The lowest relative risk (RR) was at intakes of 3.5 cups/day for all-cause mortality (RR = 0.85, 95% CI 0.82–0.89), 2.5 cups/day for CVD mortality (RR = 0.83, 95% CI 0.80–0.87), and 2 cups/day for cancer mortality (RR = 0.96, 95% CI 0.94–0.99), while additional intakes were not associated with further lower mortality. An inverse association between coffee consumption and all-cause mortality was maintained irrespective of age, overweight status, alcohol drinking, smoking status, and caffeine content of coffee. By region, Europe and Asia showed stronger inverse associations than US. A non-linear inverse association was found for mortality from respiratory disease and diabetes, while linear inverse association was found for mortality from non-CVD, non-cancer causes. Moderate coffee consumption (e.g. 2–4 cups/day) was associated with reduced all-cause and cause-specific mortality, compared to no coffee consumption. The inverse association between coffee and all-cause mortality was consistent by potential modifiers except region.

A descriptive review of variable selection methods in four epidemiologic journals: there is still room for improvement Abstract A review of epidemiological papers conducted in 2009 concluded that several studies employed variable selection methods susceptible to introduce bias and yield inadequate inferences. Many new confounder selection methods have been developed since then. The goal of the study was to provide an updated descriptive portrait of which variable selection methods are used by epidemiologists for analyzing observational data. Studies published in four major epidemiological journals in 2015 were reviewed. Only articles concerned with a predictive or explicative objective and reporting on the analysis of individual data were included. Method(s) employed for selecting variables were extracted from retained articles. A total of 975 articles were retrieved and 299 met eligibility criteria, 292 of which pursued an explicative objective. Among those, 146 studies (50%) reported using prior knowledge or causal graphs for selecting variables, 34 (12%) used change in effect estimate methods, 26 (9%) used stepwise approaches, 16 (5%) employed univariate analyses, 5 (2%) used various other methods and 107 (37%) did not provide sufficient details to allow classification (more than one method could be employed in a single article). Despite being less frequent than in the previous review, stepwise and univariable analyses, which are susceptible to introduce bias and produce inadequate inferences, were still prevalent. Moreover, 37% studies did not provide sufficient details to assess how variables were selected. We thus believe there is still room for improvement in variable selection methods used by epidemiologists and in their reporting.

Interaction between lifestyle and genetic susceptibility in myopia: the Generation R study Abstract Myopia is a refractive error of the eye caused by a complex interplay between nature and nurture. The aim of this study was to investigate whether environmental risk factors can influence the genetic effect in children developing myopia. A total of 3422 children participating in the birth-cohort study Generation R underwent an extensive eye examination at 9 years with measurements of refractive error and axial length corneal radius ratio (AL/CR). Environmental risk factors were evaluated using a questionnaire, and environmental risk scores (ERS) were calculated using backward regression analyses. Genetic risk scores (GRS) were calculated based on all currently known risk variants for myopia. Gene-environment interaction (G×E) was investigated using linear and logistic regression analyses. The predictive value of G×E and parental myopia was estimated using receiver operating characteristic curves. Myopia prevalence was 12%. Both GRS (P < 0.01) and ERS (P < 0.01) were significantly associated with myopia and AL/CR, as was G×E interaction (P < 0.01 for myopia; P = 0.07 for AL/CR). The predictive value of parental myopia was 0.67 (95% CI 0.65–0.70), similar to the values of GRS (0.67; 95% CI 0.64–0.70; P = 0.98) and ERS (0.69; 95% CI 0.66–0.72; P = 0.98). Adding G×E interaction significantly improved the predictive value to 0.73 (95% CI 0.70–0.75; P < 0.01). This study provides evidence that nature and nurture are equally important for myopia and AL/CR; however, the combination has the strongest influence. Since myopia genes are common in the population, adjustment of lifestyle should be a major focus in the prevention of myopia.

Birth seasonality and risk of autism spectrum disorder Abstract Season of birth has been hypothesized to be a risk factor for autism spectrum disorder (ASD). However, the evidence has been mixed and limited due to methodological challenges. We examine ASD birth trends for 5,464,628 births across 5 countries. ASD birth prevalence data were obtained from the International Collaboration for Autism Registry Epidemiology database, including children born in Denmark, Finland, Norway, Sweden, and Western Australia. Empirical mode decomposition and cosinor modeling were used to assess seasonality. We show seasonal variation in ASD births for the countries of Finland and Sweden. There was a modest increase in risk for children born in the fall and a modest decrease in risk for children born in the spring. Solar radiation levels around conception and the postnatal period were inversely correlated with seasonal trends in ASD risk. In the first multinational study of birth seasonality of ASD, there was evidence supporting the presence of seasonal trends in Finland and Sweden. The observations that risk was highest for fall births (i.e., conceived in the winter) and lowest for spring births (i.e., conceived in the summer), and sunlight levels during critical neurodevelopmental periods explained much of the seasonal trends, are consistent with the hypothesis that a seasonally fluctuating risk factor may influence risk of ASD.

Quantification of biological age as a determinant of age-related diseases in the Rotterdam Study: a structural equation modeling approach Abstract Chronological age alone is not a sufficient measure of the true physiological state of the body. The aims of the present study were to: (1) quantify biological age based on a physiological biomarker composite model; (2) and evaluate its association with death and age-related disease onset in the setting of an elderly population. Using structural equation modeling we computed biological age for 1699 individuals recruited from the first and second waves of the Rotterdam study. The algorithm included nine physiological parameters (c-reactive protein, creatinine, albumin, total cholesterol, cytomegalovirus optical density, urea nitrogen, alkaline phosphatase, forced expiratory volume and systolic blood pressure). We assessed the association between biological age, all-cause mortality, all-cause morbidity and specific age-related diseases over a median follow-up of 11 years. Biological age, compared to chronological age or the traditional biomarkers of age-related diseases, showed a stronger association with all-cause mortality (HR 1.15 vs. 1.13 and 1.10), all-cause morbidity (HR 1.06 vs. 1.05 and 1.03), stroke (HR 1.17 vs. 1.08 and 1.04), cancer (HR 1.07 vs. 1.04 and 1.02) and diabetes mellitus (HR 1.12 vs. 1.01 and 0.98). Individuals who were biologically younger exhibited a healthier life-style as reflected in their lower BMI (P < 0.001) and lower incidence of stroke (P < 0.001), cancer (P < 0.01) and diabetes mellitus (P = 0.02). Collectively, our findings suggest that biological age based on the biomarker composite model of nine physiological parameters is a useful construct to assess individuals 65 years and older at increased risk for specific age-related diseases.

Generalizing from the results of randomized studies of treatment: Can non-randomized studies be of help?

Intake of 12 food groups and disability-adjusted life years from coronary heart disease, stroke, type 2 diabetes, and colorectal cancer in 16 European countries Abstract Our aim was to estimate and rank 12 food groups according to disability-adjusted life years (DALYs) from coronary heart disease (CHD), stroke, type 2 diabetes (T2D), and colorectal cancer (CRC) in 16 European countries. De novo published non-linear dose–response meta-analyses of prospective studies (based on 297 primary reports), and food consumption data from the European Food Safety Authority Comprehensive European Food Consumption Database in Exposure Assessment, and DALY estimates from the Institute for Health Metrics and Evaluation were used. By implementing disease-specific counterfactual scenarios of theoretical minimum risk exposure level (TMRELs), the proportion of DALYs attributed to 12 food groups was estimated. In addition, a novel modelling approach was developed to obtain a single (optimized) TMREL across diseases. Four scenarios were analysed (A: disease-specific TMRELs/all food-disease associations; B: disease-specific TMRELs/only significant food-disease associations; C: single TMREL/all food-disease associations; D: single TMREL/only significant food-disease associations). Suboptimal food intake was associated with the following proportions of DALYs; Scenario A (highest-estimate) and D (lowest-estimate): CHD (A: 67%, D: 52%), stroke (A: 49%, D: 30%), T2D (A: 57%, D: 51%), and CRC (A: 54%, D: 40%). Whole grains (10%) had the highest impact on DALYs, followed by nuts (7.1%), processed meat (6.4%), fruit (4.4%) and fish and legumes (4.2%) when combining all scenarios. The contribution to total DALYs of all food groups combined in the different scenarios ranged from 41–52% in Austria to 51–69% in the Czech-Republic. These findings could have important implications for planning future food-based dietary guidelines as a public health nutrition strategy.

Association between tea consumption and risk of cancer: a prospective cohort study of 0.5 million Chinese adults Abstract Current experimental and epidemiological studies provide inconsistent evidence toward the association between tea consumption and cancer incidence. We investigated whether tea consumption was associated with the incidence of all cancers and six leading types of cancer (lung cancer, stomach cancer, colorectal cancer, liver cancer, female breast cancer and cervix uteri cancer) among 455,981 participants aged 30–79 years in the prospective cohort China Kadoorie Biobank. Tea consumption was assessed at baseline (2004–2008) with an interviewer-administered questionnaire. Cancer cases were identified by linkage to the national health insurance system. Cox proportional hazard regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). In the present population, daily tea consumers were more likely to be current smokers and daily alcohol consumers. 22,652 incident cancers occurred during 10.1 years follow-up (5.04 cases/1000 person-years). When we restricted analyses to non-smokers and non-excessive alcohol consumers to minimize confounding, tea consumption was not associated with all cancers (daily consumers who added tea leaves > 4.0 g/day vs. less-than-weekly consumers: HR, 1.03; 95%CI, 0.93–1.13), lung cancer (HR, 1.08; CI, 0.84–1.40), colorectal cancer (HR, 1.08; CI, 0.81–1.45) and liver cancer (HR, 1.08; CI, 0.75–1.55), yet might be associated with increased risk of stomach cancer (HR, 1.46; CI, 1.07–1.99). In both less-than-daily and daily tea consumers, all cancer risk increased with the amount of tobacco smoked or alcohol consumed. Our findings suggest tea consumption may not provide preventive effect against cancer incidence.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Industrial Microbiology & Biotechnology

Synthetic microbial consortia for biosynthesis and biodegradation: promises and challenges Abstract Functional differentiation and metabolite exchange enable microbial consortia to perform complex metabolic tasks and efficiently cycle the nutrients. Inspired by the cooperative relationships in environmental microbial consortia, synthetic microbial consortia have great promise for studying the microbial interactions in nature and more importantly for various engineering applications. However, challenges coexist with promises, and the potential of consortium-based technologies is far from being fully harnessed. Thorough understanding of the underlying molecular mechanisms of microbial interactions is greatly needed for the rational design and optimization of defined consortia. These knowledge gaps could be potentially filled with the assistance of the ongoing revolution in systems biology and synthetic biology tools. As current fundamental and technical obstacles down the road being removed, we would expect new avenues with synthetic microbial consortia playing important roles in biological and environmental engineering processes such as bioproduction of desired chemicals and fuels, as well as biodegradation of persistent contaminants.

Promoting microbial utilization of phenolic substrates from bio-oil Abstract The economic viability of the biorefinery concept is limited by the valorization of lignin. One possible method of lignin valorization is biological upgrading with aromatic-catabolic microbes. In conjunction, lignin monomers can be produced by fast pyrolysis and fractionation. However, biological upgrading of these lignin monomers is limited by low water solubility. Here, we address the problem of low water solubility with an emulsifier blend containing approximately 70 wt% Tween® 20 and 30 wt% Span® 80. Pseudomonas putida KT2440 grew to an optical density (OD600) of 1.0 ± 0.2 when supplied with 1.6 wt% emulsified phenolic monomer-rich product produced by fast pyrolysis of red oak using an emulsifier dose of 0.076 ± 0.002 g emulsifier blend per g of phenolic monomer-rich product. This approach partially mitigated the toxicity of the model phenolic monomer p-coumarate to the microbe, but not benzoate or vanillin. This study provides a proof of concept that processing of biomass-derived phenolics to increase aqueous availability can enhance microbial utilization.

Stable N -acetyltransferase Mpr1 improves ethanol productivity in the sake yeast Saccharomyces cerevisiae Abstract N-Acetyltransferase Mpr1 was originally discovered as an enzyme that detoxifies l-azetidine-2-carboxylate through its N-acetylation in the yeast Saccharomyces cerevisiae Σ1278b. Mpr1 protects yeast cells from oxidative stresses possibly by activating a novel l-arginine biosynthesis. We recently constructed a stable variant of Mpr1 (N203K) by a rational design based on the structure of the wild-type Mpr1 (WT). Here, we examined the effects of N203K on ethanol fermentation of the sake yeast S. cerevisiae strain lacking the MPR1 gene. When N203K was expressed in the diploid Japanese sake strain, its fermentation performance was improved compared to WT. In a laboratory-scale brewing, a sake strain expressing N203K produced more ethanol than WT. N203K also affected the contents of flavor compounds and organic acids. These results suggest that the stable Mpr1 variant contributes to the construction of new industrial yeast strains with improved fermentation ability and diversity of taste and flavor.

Influence of R and S enantiomers of 1-octen-3-ol on gene expression of Penicillium chrysogenum Abstract Inhibition of spore germination offers an attractive and effective target for controlling fungal species involved in food spoilage. Mushroom alcohol (1-octen-3-ol) functions as a natural self-inhibitor of spore germination for many fungi and, therefore, provides a useful tool for probing the molecular events controlling the early stages of fungal growth. In Penicillium spp., the R and S enantiomers of 1-octen-3-ol delayed spore germination and sporulation in four species of Penicillium involved in soils of fruit and grains, but to different degrees. Because of its well-annotated genome, we used Penicillium chrysogenum to perform a comprehensive comparative transcriptomic analysis of cultures treated with the two enantiomers. Altogether, about 80% of the high-quality reads could be mapped to 11,396 genes in the reference genome. The top three active pathways were metabolic (978 transcripts), biosynthesis of secondary metabolites (420 transcripts), and microbial metabolism in diverse environments (318 transcripts). When compared to the control, treatment with (R)-(-)-1-octen-3-ol affected the transcription levels of 91 genes, while (S)-(+)-1-octen-3-ol affected only 41 genes. Most of the affected transcripts were annotated and predicted to be involved in transport, establishment of localization, and transmembrane transport. Alternative splicing and SNPs' analyses indicated that, compared to the control, the R enantiomer had greater effects on the gene expression pattern of Penicillium chrysogenum than the S enantiomer. A qRT-PCR analysis of 28 randomly selected differentially expressed genes confirmed the transcriptome data. The transcriptomic data have been deposited in NCBI SRA under the accession number SRX1065226.

Metabolic engineering of glucose uptake systems in Corynebacterium glutamicum for improving the efficiency of l -lysine production Abstract Traditional amino acid producers typically exhibit the low glucose uptake rate and growth deficiency, resulting in a long fermentation time because of the accumulation of side mutations in breeding of strains. In this study, we demonstrate that the efficiency of l-lysine production in traditional l-lysine producer Corynebacterium glutamicum ZL-9 can be improved by rationally engineering glucose uptake systems. To do this, different bypasses for glucose uptake were investigated to reveal the best glucose uptake system for l-lysine production in traditional l-lysine producer. This study showed that overexpression of the key genes in PTSGlc or non-PTSGlc increased the glucose consumption, growth rate, and l-lysine production. However, increasing the function of PTSGlc in glucose uptake led to the increase of by-products, especially for plasmid-mediated expression system. Increasing the participation of non-PTSGlc in glucose utilization showed the best glucose uptake system for l-lysine production. The final strain ZL-92 with increasing the expression level of iolT1, iolT2 and ppgK could produce 201.6 ± 13.8 g/L of l-lysine with a productivity of 5.04 g/L/h and carbon yield of 0.65 g/(g glucose) in fed-batch culture. This is the first report of a rational modification of glucose uptake systems that improve the efficiency of l-lysine production through increasing the participation of non-PTSGlc in glucose utilization in traditional l-lysine producer. Similar strategies can be also used for producing other amino acids or their derivatives.

Over-expression of Isu1p and Jac1p increases the ethanol tolerance and yield by superoxide and iron homeostasis mechanism in an engineered Saccharomyces cerevisiae yeast Abstract The ethanol stress response in ethanologenic yeast during fermentation involves the swishing of several adaptation mechanisms. In Saccharomyces cerevisiae, the Jac1p and Isu1p proteins constitute the scaffold system for the Fe–S cluster assembly. This study was performed using the over-expression of the Jac1p and Isu1p in the industrially utilized S. cerevisiae UMArn3 strain, with the objective of improving the Fe–S assembly/recycling, and thus counteracting the toxic effects of ethanol stress during fermentation. The UMArn3 yeast was transformed with both the JAC1-His and ISU1-His genes-plasmid contained. The Jac1p and Isu1p His-tagged proteins over-expression in the engineered yeasts was confirmed by immunodetection, rendering increases in ethanol tolerance level from a DL50 = ~ 4.5% ethanol (v/v) to DL50 = ~ 8.2% ethanol (v/v), and survival up 90% at 15% ethanol (v/v) comparing to ~ 50% survival in the control strain. Fermentation by the engineered yeasts showed that the ethanol production was increased, producing 15–20% more ethanol than the control yeast. The decrease of ROS and free-iron accumulation was observed in the engineered yeasts under ethanol stress condition. The results indicate that Jac1p and Isu1p over-expression in the S. cerevisiaeUMArn3.3 yeast increased its ethanol tolerance level and ethanol production by a mechanism that involves ROS and iron homeostasis related to the biogenesis/recycling of Fe–S clusters dependent proteins.

Effect of applied voltage and temperature on methane production and microbial community in microbial electrochemical anaerobic digestion systems treating swine manure Abstract Microbial electrochemical technology (MET) that can harvest electricity/valuable materials and enhance the efficiency of conventional biological processes through the redox reaction of organic/inorganic compounds has attracted considerable attention. MET-based anaerobic digestion (AD) systems treating swine manure were operated at different applied voltages (0.1, 0.3, 0.5, 0.7, and 0.9 V) and temperatures (25, 35, and 45 °C). Among the MET-based AD systems with different applied voltages at 35 °C, M4 at 0.7 V showed the highest methane production (2.96 m3-CH4/m3) and methane yield (0.64 m3-CH4/kg-VS). The methane production and yield increased with increasing temperature at an applied voltage of 0.7 V. Nevertheless, the MET-based AD systems (LM at 25 °C and 0.7V) showed competitive AD performance (2.33 m3-CH4/m3 and 0.53 m3-CH4/VS) compared with the conventional AD system (35 °C). The microbial community was affected by the applied voltage and temperature, and hydrogenotrophic methanogens such as M. flavescens, M. hungatei, and M. thermautotrophicus were mainly responsible for methane production in MET-based AD systems. Therefore, the methane production can be enhanced by an applied voltage or by direct interspecies electron transfer because M. flavescens and M. thermautotrophicus were especially predominant in cathode of MET-based AD systems. The MET-based AD systems can help enhance biogas production from swine manure with no significant change in methane content. Furthermore, MET-based AD systems will be a promising AD system through low material development and the optimal operation.

Characterization and engineering of the Lrp/AsnC family regulator SACE_5717 for erythromycin overproduction in Saccharopolyspora erythraea Abstract In this work, we found that the Lrp/AsnC family protein SACE_5717 negatively regulated erythromycin biosynthesis in S. erythraea. Disruption of SACE_5717 led to a 27% improvement in the yield of erythromycin in S. erythraea A226. SACE_5717 directly repressed its own gene expression, as well as that of the adjacent gene SACE_5716 by binding to the target sequence 5′-GAACGTTCGCCGTCACGCC-3′. The predicted LysE superfamily protein SACE_5716 directly influenced the export of lysine, histidine, threonine and glycine in S. erythraea. Arginine, tyrosine and tryptophan were characterized as the effectors of SACE_5717 by weakening the binding affinity of SACE_5717. In the industrial S. erythraea WB strain, deletion of SACE_5717 (WBΔSACE_5717) increased erythromycin yield by 20%, and by 36% when SACE_5716 was overexpressed in WBΔSACE_5717 (WBΔSACE_5717/5716). In large-scale 5-L fermentation experiment, erythromycin yield in the engineered strain WBΔSACE_5717/5716 reached 4686 mg/L, a 41% enhancement over 3323 mg/L of the parent WB strain.

Inactivation of the uptake hydrogenase in the purple non-sulfur photosynthetic bacterium Rubrivivax gelatinosus CBS enables a biological water–gas shift platform for H 2 production Abstract Biological H2 production has potential to address energy security and environmental concerns if produced from renewable or waste sources. The purple non-sulfur photosynthetic bacterium Rubrivivax gelatinosus CBS produces H2 while oxidizing CO, a component of synthesis gas (Syngas). CO-linked H2 production is facilitated by an energy-converting hydrogenase (Ech), while a subsequent H2 oxidation reaction is catalyzed by a membrane-bound hydrogenase (MBH). Both hydrogenases contain [NiFe] active sites requiring 6 maturation factors (HypA-F) for assembly, but it is unclear which of the two annotated sets of hyp genes are required for each in R. gelatinosus CBS. Herein, we report correlated expression of hyp1 genes with Ech genes and hyp2 expression with MBH genes. Moreover, we find that while Ech H2 evolving activity is only delayed when hyp1 is deleted, hyp2 deletion completely disrupts MBH H2 uptake, providing a platform for a biologically driven water–gas shift reaction to produce H2 from CO.

CRISPR/Cas9-mediated engineering of Escherichia coli for n -butanol production from xylose in defined medium Abstract Butanol production from agricultural residues is the most promising alternative for fossil fuels. To reach the economic viability of biobutanol production, both glucose and xylose should be utilized and converted into butanol. Here, we engineered a dual-operon-based synthetic pathway in the genome of E. coli MG1655 to produce n-butanol using CRISPR/Cas9 technology. Further deletion of competing pathway followed by fed-batch cultivation of the engineered strain in a bioreactor with glucose-containing complex medium yielded 5.4 g/L n-butanol along with pyruvate as major co-product, indicating a redox imbalance. To ferment xylose into butanol in redox-balanced manner, we selected SSK42, an ethanologenic E. coli strain engineered and evolved in our laboratory to produce ethanol from xylose, for integrating synthetic butanol cassette in its genome via CRISPR/Cas9 after deleting the gene responsible for endogenous ethanol production. The engineered plasmid- and marker-free strain, ASA02, produced 4.32 g/L butanol in fed-batch fermentation in completely defined AM1–xylose medium.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com