Medical Specialties Commons^™

Predictive Screening Of M1 And M2 Macrophages Reveals The Immunomodulatory Effectiveness Of Post Spinal Cord Injury Azithromycin Treatment, John C. Gensel, Timothy J. Kopper, Bei Zhang, Michael B. Orr, William M. Bailey

Spinal Cord and Brain Injury Research Center Faculty Publications

Spinal cord injury (SCI) triggers a heterogeneous macrophage response that when experimentally polarized toward alternative forms of activation (M2 macrophages) promotes tissue and functional recovery. There are limited pharmacological therapies that can drive this reparative inflammatory state. In the current study, we used in vitrosystems to comprehensively defined markers of macrophages with known pathological (M1) and reparative (M2) properties in SCI. We then used these markers to objectively define the macrophage activation states after SCI in response to delayed azithromycin treatment. Mice were subjected to moderate-severe thoracic contusion SCI. Azithromycin or vehicle was administered beginning 30 minutes post-SCI and …

Identification Of Candidate Infection Genes From The Model Entomopathogenic Nematode Heterorhabditis Bacteriophora., Jonathan Vadnal, Ramesh Ratnappan, Melissa Keaney, Eric Kenney, Ioannis Eleftherianos, Damien O'Halloran, John M. Hawdon

Microbiology, Immunology, and Tropical Medicine Faculty Publications

BACKGROUND: Despite important progress in the field of innate immunity, our understanding of host immune responses to parasitic nematode infections lags behind that of responses to microbes. A limiting factor has been the obligate requirement for a vertebrate host which has hindered investigation of the parasitic nematode infective process. The nematode parasite Heterorhabditis bacteriophora offers great potential as a model to genetically dissect the process of infection. With its mutualistic Photorhabdus luminescens bacteria, H. bacteriophora invades multiple species of insects, which it kills and exploits as a food source for the development of several nematode generations. The ability to culture …

Increased Ros Production In Non-Polarized Mammary Epithelial Cells Induces Monocyte Infiltration In 3d Culture, Linzhang Li, Jie Chen, Gaofeng Xiong, Daret K. St. Clair, Wei Xu, Ren Xu

Markey Cancer Center Faculty Publications

Loss of epithelial cell polarity promotes cell invasion and cancer dissemination. Therefore, identification of factors that disrupt polarized acinar formation is crucial. Reactive oxygen species (ROS) drive cancer progression and promote inflammation. Here, we show that the non-polarized breast cancer cell line T4-2 generates significantly higher ROS levels than polarized S1 and T4R cells in three-dimensional (3D) culture, accompanied by induction of the nuclear factor κB (NF-κB) pathway and cytokine expression. Minimizing ROS in T4-2 cells with antioxidants reestablished basal polarity and inhibited cell proliferation. Introducing constitutively activated RAC1 disrupted cell polarity and increased ROS levels, indicating that RAC1 is …

Obstacles To Preventing Obesity In Children Aged 2 To 5 Years: Latino Mothers’ And Fathers’ Experiences And Perceptions Of Their Urban Environments, Carlos Penilla, Jeanne M. Tschann, Emma V. Sanchez-Vaznaugh, Elena Flores, Emily J. Ozer

School of Education Faculty Research

Background

The prevalence of obesity among Latino children is alarmingly high, when compared to non-Latino White children. Low-income Latino parents living in urban areas, even if they are well-educated, face obstacles that shape familial health behaviors. This study used qualitative methods to explore parents’ experiences in providing meals and opportunities to play to their children aged 2 to 5 years. In contrast to most prior studies, this study examined perceptions of familial behaviors among both mothers and fathers.

Methods

An ecological framework for exploring the associations of parental feeding behaviors and children’s weight informed this study. An interview guide was …

Death Related To Fistulas Involving The Gastrointestinal And Cardiovascular Systems, Allan Joseph Medwick Edd, Joseph Prahlow

Research Day

BACKGROUND/INTRODUCTION: Gastrointestinal tract fistulas are relatively uncommon and can be congenital or acquired. Fistulas may communicate with a variety of anatomic locations, including body cavities, the skin, and other organ systems. Acquired fistulas which communicate with the cardiovascular system are of particular concern, as they may be associated with significant morbidity and mortality. This study reports on three cases of acquired, post-surgical fistula formation involving the gastrointestinal and cardiovascular systems. CASE REPORTS: Case 1 - A 57-year-old man experienced sudden cardiac death shortly after having a permanent pacemaker placed for atrial fibrillation. Approximately two years earlier, he underwent surgical resection …

The Impact Of Wmed Family Medicine Obesity Clinic On Weight Loss And Patient's Overall Health In The Family Health Center, Kalamazoo, Mi, Latifa Pacheco Do, Abigail Annan Md

Research Day

No abstract provided.

Genomes Of Fasciola Hepatica From The Americas Reveal Colonization With Neorickettsia Endobacteria Related To The Agents Of Potomac Horse And Human Sennetsu Fevers., Samantha N Mcnulty, Jose F Tort, Gabriel Rinaldi, Bruce A Rosa, Victoria H. Mann, Patricia S. Latham, Paul J. Brindley, +Several Additional Authors

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Food borne trematodes (FBTs) are an assemblage of platyhelminth parasites transmitted through the food chain, four of which are recognized as neglected tropical diseases (NTDs). Fascioliasis stands out among the other NTDs due to its broad and significant impact on both human and animal health, as Fasciola sp., are also considered major pathogens of domesticated ruminants. Here we present a reference genome sequence of the common liver fluke, Fasciola hepatica isolated from sheep, complementing previously reported isolate from cattle. A total of 14,642 genes were predicted from the 1.14 GB genome of the liver fluke. Comparative genomics indicated that F. …

Fit For Genomic And Proteomic Purposes: Sampling The Fitness Of Nucleic Acid And Protein Derivatives From Formalin Fixed Paraffin Embedded Tissue., Anna Yakovleva, Jordan L Plieskatt, Sarah Jensen, Razan Humeida, Jonathan Lang, Guangzhao Li, Paige Bracci, Sylvia Silver, Jeffrey Michael Bethony

Microbiology, Immunology, and Tropical Medicine Faculty Publications

The demand for nucleic acid and protein derivatives from formalin-fixed paraffin-embedded (FFPE) tissue has greatly increased due to advances in extraction and purification methods, making these derivatives available for numerous genomic and proteomic platforms. Previously, DNA, RNA, microRNA (miRNA), or protein derived from FFPE tissue blocks were considered “unfit” for such platforms, as the process of tissue immobilization by FFPE resulted in cross-linked, fragmented, and chemically modified macromolecules. We conducted a systematic examination of nucleic acids and proteins co-extracted from 118 FFPE blocks sampled from the AIDS and Cancer Specimen Resource (ACSR) at The George Washington University after stratification by …

Schistosoma Mansoni Reinfection: Analysis Of Risk Factors By Classification And Regression Tree (Cart) Modeling., Andréa Gazzinelli, Roberta Oliveira-Prado, Leonardo Ferreira Matoso, Bráulio M Veloso, Gisele Andrade, Helmut Kloos, Jeffrey M Bethony, Renato M Assunção, Rodrigo Correa-Oliveira

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Praziquantel (PZQ) is an effective chemotherapy for schistosomiasis mansoni and a mainstay for its control and potential elimination. However, it does not prevent against reinfection, which can occur rapidly in areas with active transmission. A guide to ranking the risk factors for Schistosoma mansoni reinfection would greatly contribute to prioritizing resources and focusing prevention and control measures to prevent rapid reinfection. The objective of the current study was to explore the relationship among the socioeconomic, demographic, and epidemiological factors that can influence reinfection by S. mansoni one year after successful treatment with PZQ in school-aged children in Northeastern Minas …

Ancylostoma Ceylanicum Infective Third-Stage Larvae Are Activated By Co-Culture With Ht-29-Mtx Intestinal Epithelial Cells, Caitlin Feather, John M. Hawdon, John March

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Background

Human hookworm larvae arrest development until they enter an appropriate host. This makes it difficult to access the larvae for studying larval development or host-parasite interactions. While there are in vivo and in vitro animal models of human hookworm infection, there is currently no human, in vitro model. While animal models have provided much insight into hookworm biology, there are limitations to how closely this can replicate human infection. Therefore, we have developed a human, in vitro model of the initial phase of hookworm infection using intestinal epithelial cell culture.

Results

Co-culture of the human hookworm Ancylostoma ceylanicum with …

H-Ipse Is A Pathogen-Secreted Host Nucleus Infiltrating Protein (Infiltrin) 3 Expressed Exclusively By The Schistosoma Haematobium Egg Stage, Luke Pennington, Abdulaziz Alouffi, Evaristus Mbanefo, Debalina Ray, David Heery, Theodore Jardetzky, Michael Hsieh, Franco Falcone

Urology Faculty Publications

Urogenital schistosomiasis, caused by the parasitic trematode Schistosoma haematobium, affects over 112 million people worldwide. As with S. mansoniinfections, the pathology in urogenital schistosomiasis is mainly related to the egg stage, which induces granulomatous inflammation of affected tissues. Schistosoma eggs and their secretions have been studied extensively for the related S. mansoniorganism which is more amenable to laboratory studies. Indeed, we have shown that IPSE/alpha-1 (M-IPSE herein), a major protein secreted from S .mansoni eggs, can infiltrate host cells. Although M-IPSE function is unknown, its ability to translocate to their nucleus and bind DNA suggests a possible …

Surviving As A Cancer Survivor: A Case Report Of Radiation-Induced Autonomic Dysfunction, Daphne Sy, Mark Schauer

Research Day

While radiation and chemotherapy have achieved high rates of cure for Hodgkin lymphoma, the medical community is only now starting to characterize the long-term effects of treatment on these young cancer survivors, many decades later. Mantle radiation has been implicated in significantly increased cardiovascular risks, pulmonary fibrosis, other cancers, and thyroid disease over the patient’s lifetime. However, new syndromes are still being discovered within this population as they age. The purpose of this case study was to consider the unique health issues of cancer survivors, and how their treatment history should prompt us to broaden our differentials. Our patient presented …

Human Cytomegalovirus Interleukin-10 Enhances Matrigel Invasion Of Mda-Mb-231 Breast Cancer Cells, Cendy Valle Oseguera, Juliet Spencer

Biology Faculty Publications

Background: While some risk factors for breast cancer are well-known, the influence of other factors, particularly virus infection, remains unclear. Human cytomegalovirus (HCMV) is widespread in the general population, and both molecular and epidemiological evidence has indicated links between HCMV and breast cancer. The HCMV protein cmvIL-10 is a potent suppressor of immune function that has also been shown to promote proliferation and migration of breast cancer cells. In this study, the impact of cmvIL-10 on tumor cell invasion through a simulated basement membrane was investigated.

Results: MDA-MB-231 breast cancer cells exhibited invasion through a matrigel layer that was significantly …

Toward A Rapid Production Of Multivirus-Specific T Cells Targeting Bkv, Adenovirus, Cmv, And Ebv From Umbilical Cord Blood, Hema Dave, Min Luo, J.W. Blaney, Shabnum Patel, Cecilia Barese, Conrad Russell Cruz, Elizabeth J. Shpall, Catherine M. Bollard, Patrick J. Hanley

Pediatrics Faculty Publications

Umbilical cord blood (CB) has emerged as an effective alternative donor source for hematopoietic stem cell transplantation. Despite this success, the prolonged duration of immune suppression following CB transplantation and the naiveté of CB T cells leave patients susceptible to viral infections. Adoptive transfer of ex vivo-expanded virus-specific T cells from CB is both feasible and safe. However, the manufacturing process of these cells is complicated, lengthy, and labor-intensive. We have now developed a simplified method to manufacture a single culture of polyclonal multivirus-specific cytotoxic T cells in less than 30 days. It eliminates the need for a live virus …

Antegrade Endosteal Fibular Strut Augmentation For Periprosthetic Femur Fracture Above Stemmed Total Knee Arthroplasty, Tyler Snoap, Jason Roberts, Matthew Jaykel

Research Day

Abstract Title: ANTEGRADE ENDOSTEAL FIBULAR STRUT AUGMENTATION FOR PERIPROSTHETIC FEMUR FRACTURE ABOVE STEMMED TOTAL KNEE ARTHROPLASTY Abstract ID: 245 Abstract Category: Clinical Research Introduction: 
The management of peri-prosthetics fracture around total knee replacements is technically challenging. The options for operative fixation become more limited if the femoral component is stemmed, which often is the case after revision arthroplasty procedures. The goals of surgery for peri-prosthetic fractures above knee replacements are to provide a rigid construct to promote healing, allow immediate knee range of motion, and allow weight bearing during the healing process. 
Purpose: 
The purpose of this study is to …

Genetic Polymorphisms In Caveolin-1 Associate With Breast Cancer Risk In Chinese Han Population, M Wang, T Tian, X Ma, W Zhu, Y Guo, Z Duan, J Fan, S Lin, K Liu, Y Zheng, Q Sheng, Z J. Dai, H Peng

Biochemistry and Molecular Medicine Faculty Publications

Caveolin-1(CAV-1) was demonstrated to be a tumor suppressor gene and be implicated in the development of breast cancer (BC). Numerous potentially functional polymorphisms in CAV-1 have been identified, but their effects on BC were not clear. This case-control study aims to evaluate the relationship between CAV-1 polymorphisms and BC risk. 560 BC patients and 583 healthy controls were enrolled in the present study, all from Chinese Han population. We detected 3 single nucleotide polymorphisms (rs3807987, rs1997623, and rs7804372) in CAV-1 using the Sequenom MassARRAY method. The association between CAV-1genotypes and BC risk was assessed in six genetic …

Anti-Cancer Effects Of Tocotrienols In Nsclc, Lichchavi Raj Raj

Wayne State University Dissertations

Lung cancer is one of the leading causes of death among cancers, with non-small cell lung cancer (NSCLC) accounting for 80-85% of all lung cancers and a five-year survival rate of 5 % at stage IIIB. Delta-tocotrienol (δT) including other tocotrienol isomers has been shown to exhibit anti-tumor activity via inhibition of different signaling pathways in tumors including NSCLC. Previously we reported that δT reduced cell invasion via inhibition of the Notch-1 and NF-κb pathway.

Matrix metallopeptidase 9 (MMP9) dependent cell migration and invasion are key processes in cancer metastasis. Hence, its suppression is a promising strategy for cancer therapeutics. …

Ptpro Represses Erbb2-Driven Breast Oncogenesis By Dephosphorylation And Endosomal Internalization Of Erbb2., H Dong, L Ma, J Gan, W Lin, Rakesh Kumar, +9 Additional Authors

Biochemistry and Molecular Medicine Faculty Publications

The plasma membrane-associated tyrosine phosphatase PTPRO is frequently transcriptionally repressed in cancers and signifies poor prognosis of breast cancer patients. In this study, deletion of Ptpro inMMTV-Erbb2 transgenic mice dramatically shortened the mammary tumor latency and accelerated tumor growth due to loss of Ptpro within the breast cancer cells but not in surrounding tissue as confirmed by hetero-transplantation studies. Both in vitro and in vivo data demonstrated that the phosphatase activity was required for the inactivation of ERBB2 and its downstream signaling. PTPRO regulated the phosphorylation status of ERBB2 at Y1248. Co-immunoprecipitation and proximity ligation assay (Duolink) indicated that …

Medical Genetics And Genomic Medicine In The United States. Part 2: Reproductive Genetics, Newborn Screening, Genetic Counseling, Training, And Registries, Debra S. Regier, Carlos Ferreira, Suzanne Hart, Donald Hadley, Maximilian Muenke

Pediatrics Faculty Publications

eview of genetics in the United States with emphasis on the prenatal, metabolic, genetic counseling, and training aspects of the field.

Nasopharyngeal Microbiome Diversity Changes Over Time In Children With Asthma, Marcos Pérez-Losada, Lamia Alamri, Keith Crandall, Robert J. Freishtat

Pediatrics Faculty Publications

Background

The nasopharynx is a reservoir for pathogens associated with respiratory illnesses such as asthma. Next-generation sequencing (NGS) has been used to characterize the nasopharyngeal microbiome of infants and adults during health and disease; less is known, however, about the composition and temporal dynamics (i.e., longitudinal variation) of microbiotas from children and adolescents. Here we use NGS technology to characterize the nasopharyngeal microbiomes of asthmatic children and adolescents (6 to 18 years) and determine their stability over time.

Methods

Two nasopharyngeal washes collected 5.5 to 6.5 months apart were taken from 40 children and adolescents with asthma living in the …

Press-Pulse: A Novel Therapeutic Strategy For The Metabolic Management Of Cancer, Thomas Seyfried, George Yu, Joseph Maroon, Dominic D'Agostino

Urology Faculty Publications

Background

A shift from respiration to fermentation is a common metabolic hallmark of cancer cells. As a result, glucose and glutamine become the prime fuels for driving the dysregulated growth of tumors. The simultaneous occurrence of “Press-Pulse” disturbances was considered the mechanism responsible for reduction of organic populations during prior evolutionary epochs. Press disturbances produce chronic stress, while pulse disturbances produce acute stress on populations. It was only when both disturbances coincide that population reduction occurred.

Methods

This general concept can be applied to the management of cancer by creating chronic metabolic stresses on tumor cell energy metabolism (press disturbance) …

The Reliability Of The Navicular Drop Test And Its Transferability To Dynamic Movement, Joshua Krispin

College of Graduate Studies: Theses & Dissertations

Background: Navicular drop can be defined as the distance the navicular tuberosity moves from a standing weight bearing to a standing neutral position, as the subtalar joint moves from a relaxed position to a neutral position. Navicular drop is an important measurement for clinicians used to describe foot function, pronation, and excessive movement seen in several pathologies. Objective: The purpose of this study is to see if navicular drop is influenced by mode or speed of locomotion, if it is will there be other influences such as the forefoot and heel soft tissue, and will those factors influence those measures. …

An Rnai Screen To Identify Components Of A Polyamine Transport System, Adam J. Foley

Honors Undergraduate Theses

Polyamines, specifically putrescine, spermidine, and spermine, are small cationic molecules found in all organisms. Cells can biosynthetically make these molecules, or alternatively, they can be transported from the extracellular environment. Malignant cells have been shown to require relatively high amounts of polyamines. There is a chemotherapeutic agent, DFMO, used to block the biosynthesis of polyamines. Many malignant cells can circumvent DFMO therapy by activating their transport system. A potential solution is to simultaneously block biosynthesis and transport of polyamines. However, little is known about the polyamine transport system in higher eukaryotes.

This thesis aims to add to the basic biological …

Peripheral Huntingtin Silencing Does Not Ameliorate Central Signs Of Disease In The B6.Httq111/+ Mouse Model Of Huntington's Disease., Sydney R Coffey, Robert M Bragg, Shawn Minnig, Seth A Ament, Jeffrey P Cantle, Anne Glickenhaus, Daniel Shelnut, José M Carrillo, Dominic D Shuttleworth, Julie-Anne Rodier, Kimihiro Noguchi, C Frank Bennett, Nathan D Price, Holly B Kordasiewicz, Jeffrey B Carroll

Articles, Abstracts, and Reports

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease whose predominant neuropathological signature is the selective loss of medium spiny neurons in the striatum. Despite this selective neuropathology, the mutant protein (huntingtin) is found in virtually every cell so far studied, and, consequently, phenotypes are observed in a wide range of organ systems both inside and outside the central nervous system. We, and others, have suggested that peripheral dysfunction could contribute to the rate of progression of striatal phenotypes of HD. To test this hypothesis, we lowered levels of huntingtin by treating mice with antisense oligonucleotides (ASOs) targeting the murine …

Distinct Transcriptome Profiles Of Gag-Specific Cd8+ T Cells Temporally Correlated With The Protection Elicited By Sivδnef Live Attenuated Vaccine, Wuxun Lu, Yanmin Wu, Fangrui Ma, R. Paul Johnson, Qingsheng Li

Nebraska Center for Virology: Faculty Publications

The live attenuated vaccine (LAV) SIVmac239Δnef (SIVΔnef) confers the best protection among all the vaccine modalities tested in rhesus macaque model of HIV-1 infection. This vaccine has a unique feature of time-dependent protection: macaques are not protected at 3±5 weeks post vaccination (WPV), whereas immune protection emerges between 15 and 20 WPV. Although the exact mechanisms of the time-dependent protection remain incompletely understood, studies suggested that both cellular and humoral immunities contribute to this time-dependent protection. To further elucidate the mechanisms of protection induced by SIVΔnef, we longitudinally compared the global gene expression profiles of SIV Gag-CM9+ CD8+ (Gag-specific CD8+) …

Chloroviruses Have A Sweet Tooth, James L. Van Etten, Irina V. Agarkova, David D. Dunigan, Michela Tonetti, Cristina De Castro, Garry A. Duncan

Nebraska Center for Virology: Faculty Publications

Chloroviruses are large double-stranded DNA (dsDNA) viruses that infect certain isolates of chlorella-like green algae. They contain up to approximately 400 protein-encoding genes and 16 transfer RNA (tRNA) genes. This review summarizes the unexpected finding that many of the chlorovirus genes encode proteins involved in manipulating carbohydrates. These include enzymes involved in making extracellular polysaccharides, such as hyaluronan and chitin, enzymes that make nucleotide sugars, such as GDP-L-fucose and GDP-D-rhamnose and enzymes involved in the synthesis of glycans attached to the virus major capsid proteins. This latter process differs from that of all other glycoprotein containing viruses that traditionally use …

Tryptophan 375 Stabilizes The Outer-Domain Core Of Gp120 For Hiv Vaccine Immunogen Design, Duoyi Hu, Dane Bowder, Wenzhong Wei, Jesse Thompson, Mark A. Wilson, Shi-Hua Xiang

Nebraska Center for Virology: Faculty Publications

The outer-domain core of gp120 may serve as a better HIV vaccine immunogen than the full-length gp120 because of its greater stability and immunogenicity. In our previous report, we introduced two disulfide bonds to the outer-domain core of gp120 to fix its conformation into a CD4-bound state, which resulted in a significant increase in its immunogenicity when compared to the wild-type outer-domain core. In this report, to further improve the immunogenicity of the outer-domain core based immunogen, we have introduced a Tryptophan residue at gp120 amino acid sequence position 375 (375S/W). Our data from immunized guinea pigs indeed shows a …

First 100 Cases Of Bls First Responder A Dministered Naloxone In A Statewide Ems System, Joshua Mastenbrook, James Markman, Tyler Koedam, William Fales

Research Day

Abstract Title: FIRST 100 CASES OF BLS FIRST RESPONDER ADMINISTERED NALOXONE IN A STATEWIDE EMS SYSTEM Abstract ID: 240 Abstract Category: Clinical Research BACKGROUND: In October 2015, a midwestern state law mandated that all BLS first responder (FR) agencies be trained and equipped to administer naloxone to suspected opioid-overdose patients. Although well intentioned, several questions have arisen: does naloxone use displace emphasis on early positive pressure ventilation (PPV), is naloxone being appropriately administered, and does the mandate affect patient outcome. The purpose of this study is to evaluate the first 100 cases of FR administered naloxone (FR-naloxone) for administration appropriateness …

A Case Of Difficult-To-Treat Asthma Due To Uncontrolled Atypical Gastroesophageal Reflux, Alissa Welsh, Andrey Leonov

Research Day

Introduction: 
In uncontrolled asthma, comorbid conditions should be treated in order to achieve better asthma control. The contribution of gastro-esophageal reflux (GER) may be overlooked when its presentation is atypical. 
Case Report: 
11 year-old male with moderate persistent asthma and allergic rhinitis presented with persistent barky cough, diagnosed as spasmodic croup after bronchoscopy, esophagram, swallow study and magnified airway x-ray failed to show anatomical abnormality. Empiric proton pump inhibitor (PPI) treatment was started with improvement in cough. Patient was referred to Gastroenterology for evaluation of GER, despite lack of heartburn or vomiting. Endoscopy revealed esophageal candida infection which was treated. …

A Highly Unusual Case Of Metastatic Pancreatic Adenocarcinoma, Zachary Koehn, Lauren Piper

Research Day

Introduction: Pancreatic cancer is a rare malignancy, with nearly a 100% mortality rate. The strongest risk factor for pancreatic cancer is advanced age; the median age of diagnosis is 70. Here we discuss a case of pancreatic cancer in a 32 year-old woman. Case Description: S.R. is a 32 year-old female presenting with new-onset jaundice. Two months prior to presentation, she reported vomiting and LUQ pain, and was treated for H. pylori. Her symptoms improved, but she developed anorexia, RUQ pain, 35 lb weight loss and clay-colored diarrhea. Her medical history is negative except for 10-pack-year smoking history. Exam revealed …