Digital Commons Network^™

Role Of Mucin 19 In The Respiratory Tract, Kaitlin Mcbride

LSU Master's Theses

Human metapneumovirus (HMPV) is a negative sense, single stranded RNA virus belonging to the Pneumoviridae family, and represents an important pathogen that causes severe respiratory disease worldwide. There is currently no vaccine against HMPV, so it is important to study the aspects of the immune response induced by HMPV. Because infiltration of mucus is a hallmark of HMPV infection, it is warranted to study the role of mucus in the disease process. Mucin proteins make up the major component of mucus and can be found within the airway and lungs. Previous work from our laboratory demonstrated a high upregulation of …

Sequence Analysis Of The Potato Aphid Macrosiphum Euphorbiae Transcriptome Identified Two New Viruses, Marcella A. Texeira, Noa Sela, Hagop S. Atamian, Ergude Bao, Rita Chaudhury, Jacob Macwilliams, Jiangman He, Sophie Mantelin, Thomas Girke, Isgouhi Kaloshian

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

The potato aphid, Macrosiphum euphorbiae, is an important agricultural pest that causes economic losses to potato and tomato production. To establish the transcriptome for this aphid, RNA-Seq libraries constructed from aphids maintained on tomato plants were used in Illumina sequencing generating 52.6 million 75±105 bp paired-end reads. The reads were assembled using Velvet/Oases software with SEED preprocessing resulting in 22,137 contigs with an N50 value of 2,003bp. After removal of contigs from tomato host origin, 20,254 contigs were annotated using BLASTx searches against the non-redundant protein database from the National Center for Biotechnology Information (NCBI) as well as IntereProScan. …

The Role Of Nutrition In Influencing Mechanisms Involved In Environmentally Mediated Diseases, Bernhard Hennig, Michael C. Petriello, Mary V. Gamble, Young-Joon Surh, Laura A. Kresty, Norbert Frank, Nuchanart Rangkadilok, Mathuros Ruchirawat, William A. Suk

Superfund Research Center Faculty Publications

Human exposure to environmental contaminants such as persistent chlorinated organics, heavy metals, pesticides, phthalates, flame retardants, electronic waste and airborne pollutants around the world, and especially in Southeast Asian regions, are significant and require urgent attention. Given this widespread contamination and abundance of such toxins as persistent organic pollutants (POPs) in the ecosystem, it is unlikely that remediation alone will be sufficient to address the health impacts associated with this exposure. Furthermore, we must assume that the impact on health of some of these contaminants results in populations with extraordinary vulnerabilities to disease risks. Further exacerbating risk; infectious diseases, poverty …

Ethnicity, Climate, And House Types As Mortality Factors In The New Orleans' 1878 Yellow Fever Outbreak, Gabriele Richardson

LSU Doctoral Dissertations

The following study is first a demonstration that even incomplete health data from 1878 New Orleans’ yellow fever outbreak can be used with modern epidemiological methods and achieve useful and revealing results. Accordingly, this research examines three possible associations between New Orleans’ population exposed to yellow fever and the environment they lived in, using 4,755 yellow fever cases, derived from the New Orleans Board of Health report, Cemetery Burial records, Hospital, Asylum, and prison registers, and New Orleans obituaries. The first relationship addresses the vulnerability to yellow fever for people born in Italy, France, Great Britain, and the German Empire, …

Rapid Identification And Typing Of Mycobacterium Avium Complex Using Matrix Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (Maldi-Tof) Library And Biomarker Based Approaches, Claudia Antonika, John Dustin Loy, Raul Barletta, Michael Collins

UCARE: Research Products

Bacteria associated with the Mycobacterium avium complex (MAC) cause significant problems in animal agriculture and cause large economic losses and morbidity of livestock. Two clinically important subspecies of M. avium are M.avium ssp paratuberculosis (MAP) and M. avium ssp hominissuis (MAH). MAP is the etiologic agent of Johne’s disease, a chronic and fatal enteritis in ruminants, that has been linked to Crohn’s disease in humans.1 MAH has zoonotic potential as it is one of the leading causes of secondary infections in AIDS patients. Identification of M. avium in clinical samples is challenging as they have overlapping host ranges and clinical …

Developing Novel Therapeutics For Bacterial Lung Infections, Brandon J. Baer, Ruud Veldhuizen, Cory Yamashita

Western Research Forum

Background: Bacterial lung infections are leading causes of death worldwide. Unfortunately, increasing resistance to antibiotics and the inflammation often accompanying these infections are leading to poor outcomes despite antibiotic intervention. Complicating treatment further, the tree-like branching structure of the lung makes drug delivery to distal sites of infection difficult. Our research aims to address these challenges by developing new therapeutics and new tools to improve and assess drug delivery, bacterial killing and inflammation. Our therapy combines host defense peptides, which have been shown to kill antibiotic-resistant bacteria and down regulate inflammation, with a pulmonary vehicle, exogenous surfactant, that can improve …

Loop-Mediated Isothermal Amplification Methods For Diagnosis Of Bacterial Meningitis, Mitsuko Seki, Paul E. Kilgore, Eun Jin Kim, Makoto Ohnishi, Satoshi Hayakawa, Dong Wook Kim

Department of Pharmacy Practice

The rapid, accurate, and efficient identification of an infectious disease is critical to ensure timely clinical treatment and prevention in public health settings. In 2015, meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis was responsible for 379,200 (range: 322,700–444,700) deaths. Clinical features alone cannot determine whether bacterial meningitis is present; an analysis of cerebrospinal fluid (CSF) is essential. Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification method offering an alternative to polymerase chain reaction (PCR). LAMP-based assays for detection of three leading bacteria in CSF for diagnosis of meningitis have been established. The typing assays …

Calnexin Is Necessary For T Cell Transmigration Into The Central Nervous System, Amber M. Paul, Joanna Jung, Paul Eggleton, Alison Robinson, Jessica Wang, Nick Gutowski

Publications

In multiple sclerosis (MS), a demyelinating inflammatory disease of the CNS, and its animal model (experimental autoimmune encephalomyelitis; EAE), circulating immune cells gain access to the CNS across the blood-brain barrier to cause inflammation, myelin destruction, and neuronal damage. Here, we discovered that calnexin, an ER chaperone, is highly abundant in human brain endothelial cells of MS patients. Conversely, mice lacking calnexin exhibited resistance to EAE induction, no evidence of immune cell infiltration into the CNS, and no induction of inflammation markers within the CNS. Furthermore, calnexin deficiency in mice did not alter the development or function of the immune …

Tumorigenic Proteins Upregulated In The Mycn-Amplified Imr-32 Human Neuroblastoma Cells Promote Proliferation And Migration., Hayat Zaatiti, Jad Abdallah, Zeina Nasr, George Khazen, Anthony Sandler, Tamara J Abou-Antoun

Surgery Faculty Publications

Childhood neuroblastoma is one of the most common types of extra-cranial cancer affecting children with a clinical spectrum ranging from spontaneous regression to malignant and fatal progression. In order to improve the clinical outcomes of children with high-risk neuroblastoma, it is crucial to understand the tumorigenic mechanisms that govern its malignant behaviors. MYCN proto-oncogene, bHLH transcription factor (MYCN) amplification has been implicated in the malignant, treatment-evasive nature of aggressive, high-risk neuroblastoma. In this study, we used a SILAC approach to compare the proteomic signatures of MYCN-amplified IMR-32 and non-MYCN-amplified SK-N-SH human neuroblastoma cells. Tumorigenic proteins, including fatty-acid binding protein 5 …

Reducing The Risk Of Invasive Pathogens To Wildlife Health In The United States, Edward E. Clark Jr., Marshall Meyers, David Eldon Starling, Brent Stewart, Nathan Stone, Gary Tabor, Jeffrey S. White

National Invasive Species Council

Call to Action

In keeping with action items 4.3.1 and 4.3.2 of the 2016–2018 National Invasive Species Council (NISC) Management Plan, the Wildlife Health Task Team of the Invasive Species Advisory Committee (ISAC) was charged with: 1) identifying the major areas of vulnerability to native wildlife from the introduction and spread of invasive pathogens, and 2) making recommendations to address these vulnerabilities, including through potential changes in statute, regulation, policy, or practice of the relevant agencies.

Serum Tumour Necrosis Factor Alpha In Osteopenic And Osteoporotic Postmenopausal Females: A Cross-Sectional Study In Pakistan, Rafat Murad, Zahra Shezad, Saara Ahmed, Mussarat Ashraf, Murad Qadir, Rehana Rehman

Department of Biological & Biomedical Sciences

Objective: To compare biochemical parameters serum tumour necrosis factor alpha, calcium, magnesium, bone-specific alkaline phosphatase and vitamin D in postmenopausal women.
Methods: This cross-sectional study was carried out from June 2015 to July 2016 at Jinnah Medical and Dental College, Karachi, and comprised postmenopausal women. Bone mineral density done by dual energy X-ray absorptiometryscan categorised subjects by World Health Organisation classification into normal (T score > -1) osteopenic (T score between -1 and -2.5) and osteoporotic (T score < -2.5). Biochemical parameters like tumour necrosis alpha, calcium, magnesium, bone-specific alkaline phosphatase and vitamin D were measured by solid phase enzyme amplified sensitivity immunoassay method. SPSS 16 was used to analyse the data.
Results: Of the 146 women, 34(23%) were normal, 93(67%) were osteopenic and 19(13%) were osteoporotic. There was significant difference in mean body mass …

Gas Plasma Pre-Treatment Increases Antibiotic Sensitivity And Persister Eradication In Methicillin-Resistant Staphylococcus Aureus, Li Guo, Ruobing Xu, Yiming Zhao, Dingxin Liu, Zhijie Liu, Xiaohua Wang, Hailan Chen, Michael G. Kong

Bioelectrics Publications

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of serious nosocomial infections, and recurrent MRSA infections primarily result from the survival of persister cells after antibiotic treatment. Gas plasma, a novel source of ROS (reactive oxygen species) and RNS (reactive nitrogen species) generation, not only inactivates pathogenic microbes but also restore the sensitivity of MRSA to antibiotics. This study further found that sublethal treatment of MRSA with both plasma and plasma-activated saline increased the antibiotic sensitivity and promoted the eradication of persister cells by tetracycline, gentamycin, clindamycin, chloramphenicol, ciprofloxacin, rifampicin, and vancomycin. The short-lived ROS and RNS generated by plasma …

Central Nervous System Histoplasmosis: Multicenter Retrospective Study On Clinical Features, Diagnostic Approach And Outcome Of Treatment, Joseph Wheat, Thein Myint, Ying Guo, Phebe Kemmer, Chadi A. Hage, Colin Terry, Marwan M. Azar, James Riddell, Peter Ender, Sharon Chen, Kareem Shehab, Kerry Cleveland, Eden Esguerra, James Johnson, Patty Wright, Vanja Douglas, Pascalis Vergidis, Winnie Ooi, John Baddley, David Bamberger, Raed N. Khairy, Holenarasipur R. Vikram, Elizabeth Jenny-Avital, Geetha Sivasubramanian, Karen Bowlware, Barbara Pahud, Juan Sarria, Townson Tsai, Maha Assi, Satish Mocherla

Internal Medicine Faculty Publications

Central nervous system (CNS) involvement occurs in 5 to 10% of individuals with disseminated histoplasmosis. Most experience has been derived from small single center case series, or case report literature reviews. Therefore, a larger study of central nervous system (CNS) histoplasmosis is needed in order to guide the approach to diagnosis, and treatment.

A convenience sample of 77 patients with histoplasmosis infection of the CNS was evaluated. Data was collected that focused on recognition of infection, diagnostic techniques, and outcomes of treatment.

Twenty nine percent of patients were not immunosuppressed. Histoplasma antigen, or anti-Histoplasma antibodies were detected in the …

Barriers To Addressing Hiv-Associated Neurocognitive Disorder (Hand): Community-Based Service Provider Perspectives, Renato M. Liboro, Francisco Ibanez-Carrasco, Sean B. Rourke, Andrew Easton, Claudia Medina, Daniel Pugh, Allan Rae, Lori E. Ross, Paul A. Shuper

Psychology Faculty Research

HIV-Associated Neurocognitive Disorder (HAND) is an emergent public health issue in developed countries. Consequently, people living with HIV who experience HAND will increasingly require support from community-based HIV service providers. The objective of our qualitative study was to identify barriers service providers face in addressing HAND among people living with HIV. Thirty-three providers from 22 AIDS service organizations across Ontario, Canada, were interviewed. Using thematic analysis, three types of barriers were identified: (a) personal/professional, (b) service access, and (c) systemic. This paper draws attention to HAND-related obstacles that service providers encounter in their work and presents options to overcome them.

Drug Repurposing For Schistosomiasis: Combinations Of Drugs Or Biomolecules., Maria João Gouveia, Paul J Brindley, Fátima Gärtner, José M Correia Da Costa, Nuno Vale

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Schistosomiasis is a major neglected tropical disease. Control of schistosomiasis currently relies on a single drug, praziquantel, and despite its efficacy against the all schistosome species that parasitize humans, it displays some problematic drawbacks and alone is ineffective in counteracting adverse pathologies associated with infection. Moreover, due to the development of the potential emergence of PZQ-resistant strains, the search for additional or alternative antischistosomal drugs have become a public health priority. The current drug discovery for schistosomiasis has been slow and uninspiring. By contrast, repurposing of existing approved drugs may offer a safe, rapid and cost-effective alternative. Combined treatment with …

An Aged Canid With Behavioral Deficits Exhibits Blood And Cerebrospinal Fluid Amyloid Beta Oligomers, Clare Rusbridge, Francisco J. Salguero, Monique Antoinette David, Kiterie M. E. Faller, Jose T. Bras, Rita J. Guerreiro, Angela C. Richard-Londt, Duncan Grainger, Elizabeth Head, Sebastian G. P. Brandner, Brian Summers, John Hardy, Mourad Tayebi

Pharmacology and Nutritional Sciences Faculty Publications

Many of the molecular and pathological features associated with human Alzheimer disease (AD) are mirrored in the naturally occurring age-associated neuropathology in the canine species. In aged dogs with declining learned behavior and memory the severity of cognitive dysfunction parallels the progressive build up and location of Aβ in the brain. The main aim of this work was to study the biological behavior of soluble oligomers isolated from an aged dog with cognitive dysfunction through investigating their interaction with a human cell line and synthetic Aβ peptides. We report that soluble oligomers were specifically detected in the dog's blood and …

Etiology Of Community Acquired Clostridium Difficile-Associated Disease, Shannan Sherman, Dr. Pamela Ark

The Pegasus Review: UCF Undergraduate Research Journal

A review of literature related to community acquired Clostridium difficile -associated disease (CA-CDAD) was conducted. Nine relevant studies were identified using the Cumulative Index to Nursing and Allied Health (CINAHL) and MEDLINE-EBSCOhost databases. Clinical practice recommendations were obtained from the Centers for Disease Control and Prevention. The studies provided information about epidemiology of infection due to C. difficile in the community and interventions to reduce transmission. Multiple studies found underlying gastrointestinal disorders and use of cephalosporin antibiotics to be a risk factor. Another risk factor was administration of gastric acid suppressive drugs. No particular C. difficile strain was more likely …

Sertraline, Paroxetine, And Chlorpromazine Are Rapidly Acting Anthelmintic Drugs Capable Of Clinical Repurposing., Janis C Weeks, William M Roberts, Caitlyn Leasure, Brian M Suzuki, Kristin J Robinson, John M Hawdon, +Several Additional Authors

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Parasitic helminths infect over 1 billion people worldwide, while current treatments rely on a limited arsenal of drugs. To expedite drug discovery, we screened a small-molecule library of compounds with histories of use in human clinical trials for anthelmintic activity against the soil nematode Caenorhabditis elegans. From this screen, we found that the neuromodulatory drugs sertraline, paroxetine, and chlorpromazine kill C. elegans at multiple life stages including embryos, developing larvae and gravid adults. These drugs act rapidly to inhibit C. elegans feeding within minutes of exposure. Sertraline, paroxetine, and chlorpromazine also decrease motility of adult Trichuris muris whipworms, prevent hatching …

Frequency And Factors Associated With Adult Immunization In Patients Visiting Family Medicine Clinics At A Tertiary Care Hospital, Karachi, Samar Zaki, Asma Usman, Swaleha Tariq, Sameena Shah, Iqbal Azam Syed, Waris Qidwai, Kashmira Nanji

Department of Family Medicine

Objective: The goal of this study was to determine the frequency and factors associated with adult immunization in patients visiting family medicine clinics at a tertiary care hospital in Karachi.
Methods: A cross-sectional study was conducted from March 2014 to March 2015 in a tertiary care hospital in Karachi, Pakistan. Participants more than 18 years were invited to participate in the study. A pretested questionnaire was used to collect information. Data were entered and analyzed using IBM SPSS Statistics for Windows, version 19.0 (Armonk, NY: IBM Corp).
Results: A total of 340 patients were surveyed. The majority of patients …

Factors Associated With Perceived Accuracy Of The Undetectable = Untransmittable Slogan Among Men Who Have Sex With Men: Implications For Messaging Scale-Up And Implementation, H Jonathon Rendina, J T. Parsons

Publications and Research

Introduction: Recent research has shown high efficacy of HIV treatment for reducing the risk of HIV transmission to sexual partners. As the efficacy of treatment as prevention (TasP) has proliferated, a new messaging campaign, Undetectable = Untransmittable, has been gaining popularity. The purpose of this paper was to assess factors associated with the perceived accuracy of this TasP messaging strategy among a large and diverse sample of gay, bisexual, and other men who have sex with men (GBMSM) in order to inform subsequent efforts at large-scale and implementation of the HIV prevention message.

Methods: We conducted a nationwide survey of …

Granulin Secreted By The Food-Borne Liver Fluke, Brandon Haugen, Shannon E Karinshak, Victoria H Mann, Anastas Popratiloff, Alex Loukas, Paul J Brindley, Michael J Smout

Microbiology, Immunology, and Tropical Medicine Faculty Publications

The liver fluke Opisthorchis viverrini is a food-borne, zoonotic pathogen endemic to Thailand and adjacent countries in Southeast Asia. The adult developmental stage of the O. viverrini parasite excretes and secretes numerous proteins within the biliary tract including the gall bladder. Lesions caused by the feeding activities of the liver fluke represent wounds that undergo protracted cycles of healing and re-injury during chronic infection, which can last for decades. Components of the excretory/secretory (ES) complement released by the worms capably drive proliferation of bile duct epithelial cells and are implicated in establishing the oncogenic milieu that leads to bile duct …

Granulin Secreted By The Food-Borne Liver Fluke Opisthorchis Viverrini Promotes Angiogenesis In Human Endothelial Cells, Brandon Haugen, Shannon Karinshak, Victoria H. Mann, Anastas Popratiloff, Alex Loukas, Paul J. Brindley, Michael J. Smout

Microbiology, Immunology, and Tropical Medicine Faculty Publications

The liver fluke Opisthorchis viverrini is a food-borne, zoonotic pathogen endemic to Thailand and adjacent countries in Southeast Asia. The adult developmental stage of the O. viverrini parasite excretes and secretes numerous proteins within the biliary tract including the gall bladder. Lesions caused by the feeding activities of the liver fluke represent wounds that undergo protracted cycles of healing and re-injury during chronic infection, which can last for decades. Components of the excretory/secretory (ES) complement released by the worms capably drive proliferation of bile duct epithelial cells and are implicated in establishing the oncogenic milieu that leads to bile duct …

Controlled Human Hookworm Infection: Accelerating Human Hookworm Vaccine Development, David Diemert, Doreen Campbell, Jill Brelsford, Caitlyn Leasure, Guangzhao Li, Naji Younes, +Several Additional Authors

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Background

Controlled human hookworm infection (CHHI) is a central component of a proposed hookworm vaccination-challenge model (HVCM) to test the efficacy of candidate vaccines. Critical to CHHI is the manufacture of Necator americanus infective larvae (NaL3) according to current Good Manufacturing Practice (cGMP) and the determination of an inoculum of NaL3 that is safe and reliably induces patent infection. Methods

cGMP-grade NaL3 were produced for a phase 1 trial in 20 healthy, hookworm-naïve adults in the United States, who received either 25 or 50 NaL3. Participants were monitored for 12–18 weeks postinfection for safety, tolerability, …

Transplantation Of Schistosome Sporocysts Between Host Snails: A Video Guide., Gabriel Mouahid, Anne Rognon, Ronaldo De Carvalho Augusto, Patrick Driguez, Kathy Geyer, Shannon Karinshak, Nelia Luviano, Victoria Mann, Thomas Quack, Kate Rawlinson, George Wendt, Christoph Grunau, Hélène Moné

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Schistosomiasis is an important parasitic disease, touching roughly 200 million people worldwide. The causative agents are different Schistosoma species. Schistosomes have a complex life cycle, with a freshwater snail as intermediate host. After infection, sporocysts develop inside the snail host and give rise to human dwelling larvae. We present here a detailed step-by-step video instruction in English, French, Spanish and Portuguese that shows how these sporocysts can be manipulated and transferred from one snail to another. This procedure provides a technical basis for different types of ex vivo modifications, such as those used in functional genomics studies.

Lebanon: Cholera Outbreak, Muhammad Naqvi

Global Public Health

Lebanon is a country that has always been tangled with the ever looming threat of a growing health crisis. With a failing government and massive influx of refugees, mostly the victim of their war torn economy, it seemed inevitable that sanitation procedures would run short. Now the Lebanese people and government face the challenge of the spread of Cholera, a water-borne disease, within the countries vicinity as a result of mass pollution from the formation of informal settlements as well as the country being a hotbed for political instability. These have resulted in limited success in controlling the outbreak engulfing …

Guinea : Malaria, Madeleine Jacobson

Global Public Health

This brief analyzes malaria in Guinea and its impacts on individuals and the society. It consists of a geographic and demographic presentation of Guinea which includes information about its location, climate, natural resources, ethnic groups, politics and social internal and external challenges where human trafficking, lack of healthcare and refugees are discussed. The brief analyzes malaria´s transmission, diagnosis, preventions and a suggestion for an intervention.

Malaria is a vector-borne disease, which means that living organisms that can transmit infectious diseases between humans or from animals to humans, spread malaria. 70 percent of all malaria deaths occur in age years, but …

Bulgaria: Brucellosis, Austin Wheeler

Global Public Health

In this brief we will discuss how a disease called Brucellosis has affected the country of Bulgaria. Bulgaria is a European country located towards the South Eastern part of Europe. A few points mentioned in the brief are what the disease can do to your body, through education of the disease we can show what a serious problem it is. The brief will also cover how you can contract the disease through animals, and how you can prevent contraction from occurring. The brief also includes a lot about the religious background of the country. This should hopefully be used to …

The Gambia : Tuberculosis, Amen Belayneh

Global Public Health

Tuberculosis in the Gambia

This brief consists of some general information about Gambia, demographic information, the effects of tuberculosis in the country including the challenges brought forth by the disease, and some personal suggestions on the matter. From the general information on Gambia, we learn the location, political structure as well as major internal and external challenges faced by the country. Tuberculosis is one of the country’s major internal challenges in addition to its environmental and economic issues. Under the demographic information we get a broader idea of the Gambia people by looking at population demographic such as birth and …

Comoros: Malaria, Elizabeth Rennolds

Global Public Health

After human settlement and migration took place throughout the world, a disease known as malaria broke out within many countries. Malaria is a disease that comes from a fever where a parasite takes over one’s red blood cells with mosquitoes being the ones that transmit this disease. The symptoms of this disease are as follows: fever (typical cycles of fever, chills, sweating), joint pain, headaches, vomiting, convulsions, and coma. Malaria is a big issue in the Comoros, an island which can be seen in the Indian Ocean between the eastern coast of Africa and Madagascar, because of the many occurrences …

Uganda: Malaria, Sui Hnem

Global Public Health

Malaria is an infectious disease caused by mosquito bite. It is a major health problem in Uganda and is a major cause of mortality and morbidity. Pregnant women and children are the most infected groups in the country. The Ugandan government did their best to keep children and women healthy by providing clinics near the villages and advocating home treatment for malaria. Although, the government is very supportive in preventing and promoting the malaria, people did not seek healthcare as much as they should. The reason for not seeking healthcare was because they feel healthy, or the clinic had long …