1ScIeNtIFIc REPORTS | (2018) 8:1250 | DOI:10.1038/s41598-018-19638-x
www.nature.com/scientificreports
Histology, immunohistochemistry,
and in situ hybridization reveal
overlooked Ebola virus target
tissues in the Ebola virus disease
guinea pig model
Timothy K. Cooper1, Louis Huzella 1, Joshua C. Johnson 1, Oscar Rojas1, Sri Yellayi1,3,
Mei G. Sun2, Sina Bavari2, Amanda Bonilla1, Randy Hart1, Peter B. Jahrling 1, Jens H. Kuhn 1
& Xiankun Zeng 2
Survivors of Ebola virus infection may become subclinically infected, but whether animal models
recapitulate this complication is unclear. Using histology in combination with immunohistochemistry
and in situ hybridization in a retrospective review of a guinea pig confirmation-of-virulence study, we
demonstrate for the first time Ebola virus infection in hepatic oval cells, the endocardium and stroma of
the atrioventricular valves and chordae tendinae, satellite cells of peripheral ganglia, neurofibroblasts
and Schwann cells of peripheral nerves and ganglia, smooth muscle cells of the uterine myometrium
and vaginal wall, acini of the parotid salivary glands, thyroid follicular cells, adrenal medullary cells,
pancreatic islet cells, endometrial glandular and surface epithelium, and the epithelium of the vagina,
penis and, prepuce. These findings indicate that standard animal models for Ebola virus disease are not
as well-described as previously thought and may serve as a stepping stone for future identification of
potential sites of virus persistence.
Ebola virus disease (EVD) is a severe and frequently lethal affliction of humans caused by infection with any
of three members of the mononegavirus family Filoviridae: Bundibugyo virus (BDBV), Ebola virus (EBOV),
and Sudan virus (SUDV). A fourth virus, Taï Forest virus (TAFV), has thus far caused only a single reported
human infection, which was nonlethal1. EVD is an exotic disease with case numbers rarely surpassing the lower
hundreds1; however, from 2013–2016, EBOV caused an EVD outbreak in Western Africa encompassing 28,616
infections and 11,310 deaths in Guinea, Liberia, and Sierra Leone2. Long term sequelae in individual survivors of
acute EVD and the similar Marburg virus disease (MVD) and filovirus persistence followed by disease relapse or
sexual transmission had been reported before this outbreak3–8. However, observations during and following the
Western African EVD outbreak suggest that sequelae and filovirus persistence may be common events9. Reported
sequelae include arthralgia, cardiac valvulopathy, parotid gland inflammation, peripheral paresthesia or dyses-
thesia, and gastrointestinal motility disorders10–14. Semen may contain detectable EBOV RNA for more than 500
days following recovery, and EBOV RNA has been detected in breast milk of a subclinically infected mother15,16.
Replicating EBOV has been isolated from the cerebrospinal fluid of an EVD survivor suffering a disease relapse
and from the aqueous hu.
Polio: flaccid paralysis, major and minor
disease, fecal-oral
Coxsackievirus A: vesicular diseases,
meningitis; coxsackievirus B (body):
pleurodynia, myocarditis
Other echovirus and enteroviruses: like
coxsackievirus
Rhinoviruses: common cold, acid labile, does
not replicate above 33° C
Biology, Virulence, and Disease
• Small size, icosahedral capsid, positive RNA
genome with terminal protein
• Genome is sufficient for infection
• Encodes RNA-dependent RNA polymerase,
replicates in cytoplasm
Enteroviruses
• Capsid virus resistant to inactivation
• Disease due to lytic infection of important
target tissue
• Polio: cytolytic infection of motor neurons of
anterior horn and brainstem, paralysis
• Coxsackievirus A: herpangina, hand-foot-
and-mouth disease, common cold,
meningitis
• Coxsackievirus B: pleurodynia, neonatal
myocarditis, type 1 diabetes
Rhinoviruses
• Acid labile and cannot replicate at body
temperature
• Restricted to upper respiratory tract
• Common cold
Epidemiology
• Enteroviruses transmitted by fecal-oral route
and aerosols
• Rhinoviruses transmitted by aerosols and
contact
Diagnosis
• Immune assays (ELISA) or RT-PCR genome
analysis of blood, CSF, or other relevant
sample
Treatment, Prevention, and Control
• OPV and IPV polio vaccines
P
icornaviridae is one of the largest families of viruses and
includes some of the most important human and animal
viruses (Box 46-1). As the name indicates, these viruses are
small (pico) ribonucleic acid (RNA) viruses that have a
naked capsid structure. The family has more than 230
members divided into nine genera, including Enterovirus,
Rhinovirus, Hepatovirus (hepatitis A virus; discussed in
Chapter 55), Cardiovirus, and Aphthovirus. The enterovi-
ruses are distinguished from the rhinoviruses by the stabil-
ity of the capsid at pH 3, the optimum temperature
for growth, the mode of transmission, and their diseases
A total number of 74 coagulase negative Staphylococci were isolated from orthopaedic patients in Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. They were further characterized into various Staphylococci species using API STAPH identification kit: Staph xylosus (31.1%), Staph lentus (10.8%), Staph hominis (10.8%), Staph cohnii cohnii (5.4%), Staph epidermidis (4.1%) others were Staph cohnii ureal., Staph hyicus, Staph lugdunensis (2.7% each) Staph caprae , Staph capitis, Staph haemolyticus, Staph scuiri, Staph chromogenes and Staph warneri (1.4% each). Microcossus spp was 8.2% while 13.5% isolates were undetermined. Kirby Baurer disk method was used for the antibiotics susceptibility test, the result showed gentamicin and ciprofloxacin to be most active (96.6%), followed by vancomycin (93.1) and pefloxacin (87.9). The isolates were resistant to ampicillin (96.6), amoxicillin clavulanic acid (65.5%), clindamycin 41.4%). The aim of this study is to classify the coagulase negative Staphylococci isolates into species and to determine their antibiotic susceptibility
Characterization of Mycobacterium Tuberculosis Complex Strains: A Multicenter...CrimsonpublishersCJMI
Characterization of Mycobacterium Tuberculosis Complex Strains: A Multicenter Retrospective Greek Study by Lamprini Gkaravela in Cohesive Journal of Microbiology & Infectious Disease
Polio: flaccid paralysis, major and minor
disease, fecal-oral
Coxsackievirus A: vesicular diseases,
meningitis; coxsackievirus B (body):
pleurodynia, myocarditis
Other echovirus and enteroviruses: like
coxsackievirus
Rhinoviruses: common cold, acid labile, does
not replicate above 33° C
Biology, Virulence, and Disease
• Small size, icosahedral capsid, positive RNA
genome with terminal protein
• Genome is sufficient for infection
• Encodes RNA-dependent RNA polymerase,
replicates in cytoplasm
Enteroviruses
• Capsid virus resistant to inactivation
• Disease due to lytic infection of important
target tissue
• Polio: cytolytic infection of motor neurons of
anterior horn and brainstem, paralysis
• Coxsackievirus A: herpangina, hand-foot-
and-mouth disease, common cold,
meningitis
• Coxsackievirus B: pleurodynia, neonatal
myocarditis, type 1 diabetes
Rhinoviruses
• Acid labile and cannot replicate at body
temperature
• Restricted to upper respiratory tract
• Common cold
Epidemiology
• Enteroviruses transmitted by fecal-oral route
and aerosols
• Rhinoviruses transmitted by aerosols and
contact
Diagnosis
• Immune assays (ELISA) or RT-PCR genome
analysis of blood, CSF, or other relevant
sample
Treatment, Prevention, and Control
• OPV and IPV polio vaccines
P
icornaviridae is one of the largest families of viruses and
includes some of the most important human and animal
viruses (Box 46-1). As the name indicates, these viruses are
small (pico) ribonucleic acid (RNA) viruses that have a
naked capsid structure. The family has more than 230
members divided into nine genera, including Enterovirus,
Rhinovirus, Hepatovirus (hepatitis A virus; discussed in
Chapter 55), Cardiovirus, and Aphthovirus. The enterovi-
ruses are distinguished from the rhinoviruses by the stabil-
ity of the capsid at pH 3, the optimum temperature
for growth, the mode of transmission, and their diseases
A total number of 74 coagulase negative Staphylococci were isolated from orthopaedic patients in Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. They were further characterized into various Staphylococci species using API STAPH identification kit: Staph xylosus (31.1%), Staph lentus (10.8%), Staph hominis (10.8%), Staph cohnii cohnii (5.4%), Staph epidermidis (4.1%) others were Staph cohnii ureal., Staph hyicus, Staph lugdunensis (2.7% each) Staph caprae , Staph capitis, Staph haemolyticus, Staph scuiri, Staph chromogenes and Staph warneri (1.4% each). Microcossus spp was 8.2% while 13.5% isolates were undetermined. Kirby Baurer disk method was used for the antibiotics susceptibility test, the result showed gentamicin and ciprofloxacin to be most active (96.6%), followed by vancomycin (93.1) and pefloxacin (87.9). The isolates were resistant to ampicillin (96.6), amoxicillin clavulanic acid (65.5%), clindamycin 41.4%). The aim of this study is to classify the coagulase negative Staphylococci isolates into species and to determine their antibiotic susceptibility
Characterization of Mycobacterium Tuberculosis Complex Strains: A Multicenter...CrimsonpublishersCJMI
Characterization of Mycobacterium Tuberculosis Complex Strains: A Multicenter Retrospective Greek Study by Lamprini Gkaravela in Cohesive Journal of Microbiology & Infectious Disease
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
A Case of Postmenopausal Pyometra Caused By Endometrial Tuberculosisiosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Asymptomatic urinary tract infection amongst some Students of Michael Okpara ...Premier Publishers
This work was carried out to determine Asymptomatic Urinary Tract Infection amongst some students of Michael Okpara University of Agriculture, Umudike and the sensitivity pattern of the isolates from urine. Using aseptic technique, midstream urine were collected from sixty (60) students, urinalysis was carried out on the urine samples and was then cultured on CLED and MacConkey agar using pour plate method. Growth was observed in 26 (87%) of the sample while there was no growth in 4 (13%) of the sample. Out of the 26 (87%) samples with growth, 14 (47%) had significant bacteria growth while 12 (40%) had no significant growth. Incidence of asymptomatic bacteriuria was higher in females 8 (57%) than males 6 (38%). The organisms isolated were Escherichia coli, Klebsiella species, Staphylococcus saprophyticus, Staphylococcus aureus, Enterococcus faecalis, Proteus species, and Pseudomonas aeruginosa. All the Gram positive isolates were sensitive to Gentamycin and all resistance to Cefuroxime, Ceftazidime, Ceftriaxone, Cloxacillin. The Gram negative isolates were mostly sensitive to Nitrofurantoin, Gentamycin and Ofloxacin. Therefore, these drugs could be considered as the first line of drug for the treatment of asymptomatic urinary tract infection.
Ascaris lumbricoides and other Gastrointestinal Helminthic Parasites among Qe...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
FOURNIER’S GANGRENE: REVIEW OF 57 CASES IN TERTIARY INSTITUTIONAnil Haripriya
Fournier’s gangrene which is a rapidly progressive, fulminant polymicrobial synergistic infection of the perineum and genitals is now changing its pattern. Both genders can be affected and the mortality is still high (around10%). The clinical presentation in many patients in early stage may not be prominent. Thus rapid and accurate diagnosis is must for prompt treatment. Extensive surgical debridement and broad spectrum intravenous antibiotic remains the mainstay of treatment in order to reduce the morbidity and mortality.
Clinical Manifestations of Plasmodium bergheiANKA Infection in Juvenile Mice:...AI Publications
Malaria is an important health and development challenge in Africa, Animalmodels most particularly mice, have long been employedto study malaria pathogenesis. Clinical manifestations due to Plasmodium bergheiANKA infection in juvenile mice as a model for understanding the complications ofcongenital malaria in neonates.Forty-five juvenile mice (5-7 days old) were acquired from University College Hospital, Ibadan and injected with 2 x 107 (0.2ml) Plasmodium berghei ANKA parasitized red blood cells (PRBCs). Mice were transported to the study site, kept in well ventilated cages and fed daily with a balanced ration. Every day after post-P. berghei infection, mice were monitored for mortality. Clinical manifestations ofexperimental cerebral malaria (ECM) was assessed and confirmed if at leastruffled fur, hunching, wobbly gait, limb paralysis, convulsions, or coma was observed. Each sign was given a score of 1. Animals with scores ≥4 were considered to have severe ECM.20 (44%) micewerelost due to natural cause (i.e. stress) at day 2 of the experiment. Between day 4 and 9, 25 (56%) of the studymice presented clinical signs of ECM which includes; ruffled fur 25(100%), hunching 21 (84%), wobbly gait 17 (68%), limb paralysis 20 (80%), convulsions 25 (100%) and subsequently died. Survival rate and severity of ECM in the mice differs, 22 (88.0%) had severe ECM and 3(12.0%) had mild ECM.This study has shown that parasite establishment and malaria complications can manifest as early as 4 days’postP. berghei infection in 5-7 days old mice.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
The prevalence of Escherichia coli Causing Urinary Tract Infections in Aminu ...Premier Publishers
Urinary Tract infections (UTIs) are one of the most common causes of hospital visit worldwide. The study intends to find the prevalence of Escherichia coli in Urinary tract infection cases in Aminu Kano Teaching Hospital (AKTH), Kano. A comprehensive study was conducted on E coli for its prevalence in urine samples, of queried cases of urinary tract infection in patients attending Aminu Kano Teaching Hospital (AKTH), Kano. Two hundred and fourteen urine specimens comprising of 123(57.5%) females and 91(42.5%) males of all age group were screened for bacteria. Of the 214 samples, 68 representing (31.80%) were culture positive with E. coli having 32(47.06%), Klebsiella spp 18(26.47%), Staphylococcus spp 10(14.70%), Proteus spp 6(8.82%) and Pseudomonas spp 2(2.94%). Out of the 32(47.6%) of E. coli isolated, 22 were from female and 10 were from males, the highest prevalence occurred within the age range of 21-30 with females taking the lead. Antibiogram of the isolated E. coli showed a markedly good sensitivity of ofloxacin (96.87%), nitrofurantoin and nalidixic acid (93.70%), colistin phosphate (90.62%) and gentamicin (68.75%). E. coli showed the least sensitivity to ampicillin (18.75%). The frequency of E. coli obtained suggests its high prevalence. And this can be reduced using Flouroquinolones as exhibited by the susceptibility profile in this study.
COLLEGE
PHYSICS LAB REPORT
STUDENTS NAME
ANALYSIS OF A BUBBLE CHAMBER PICTURE
SUPERVISED BY:
19/05/2020
1. Introduction
A bubble chamber is a vessel filled with a superheated transparent liquid (most often liquid hydrogen) used to detect electrically charged particles moving through it. It was invented in 1952 by Donald A. Glaser, for which he was awarded the 1960 Nobel Prize in Physics.
A convenient way to study the properties of the fundamental subatomic particles is through observation of their bubble trails, or tracks, in a bubble chamber. Using measurements made directly on a bubble chamber photograph, we can often identify the particles from their tracks and calculate their masses and other properties. In a typical experiment, a beam of a particular type of particle is sent from an accelerator into a bubble chamber, which is a large liquid-filled vessel. To simplify the analysis of the data, the liquid used is often hydrogen, the simplest element. The use of liquid hydrogen, while it simplifies the analysis, complicates the experiment itself, since hydrogen, a gas at room temperature, liquefies only when cooled to -246◦C. For charged particles to leave tracks in passing through the chamber, the liquid must be in a “super-heated” state, in which the slightest disturbance causes boiling to occur. In practice, this is accomplished by expanding the vapor above the liquid with a piston a few thousandths of a second before the particles enter the chamber.
2. Methods
2.1 Materials needed:
1. student worksheet per student
2. Ruler
3. Scissors
4. Glue stick
5. Pocket calculator
2.2 Procedures
2.2.1 Calculation of the X Particle’s Mass.
Make measurements on each of the photographs. In particular, for each of the circled events measure these four quantities:
· `Σ - The length of the Σ track,
· θ - the angle between the Σ− and π− track,
· s - the sagitta of the π− track,
· `π - The chord length of the π− track.
Your values for the event should be close to those given in the sample input. Run the program using each set of measurements, and tabulate the computed X0 mass from each event. Compute an average of the calculated masses and find the average deviation, expressing your result as Mx ±∆Mx.
Compare your final result with some known neutral particles listed below and identify the X0 particle based on this comparison.
Particlemass (in MeV/c2)
π0 135
K0 498
n 940
Λ0 1116
Σ0 1192
Ξ0 1315
2.2.2 Determination of the Angle θ.
The angle θ between the π− and Σ− momentum vectors can be determined by drawing tangents to the π− and Σ− tracks at the point of the Σ− decay.
We can then measure the angle between the tangents using a protractor. We can show.
Collins did not understand the events that led to the reasoning .docxvickeryr87
Collins did not understand the events that led to the reasoning why and actions how Wells Fargo conveyed victories, achievements, and accomplishments. Therefore, Collins was incorrect. Wells Fargo as a company was able to achieve earning because of their successful strategies in profit revenue or turnover for each employee. The Good to Great article brought up that many people were surprised when finding out what made companies successful especially in relation to the leadership (Collins, 2001).
Our text states that it is difficult for companies to find good ethical leaders because they tend to be more expensive (Meese, Ortmeier, 2001). Unethical leadership is sure to bring the downfall of any business; it is impossible to have a successful business if there are unethical practices, the employees are also bound to begin these practices making it difficult for the company to hold their end of their promises to their customers or members.
"He who oppresses the poor to make more for himself or who gives to the rich, will only come to poverty" (Proverbs 22:16).
Collins, J. (2001). Good to great: Why some companies make the leap and others don't. New York, NY: Harper Collins.
Mees, E., Ortmeier, P. (2001). Leadership, ethics, and policing:Challenges for the 21st century. Upper Saddle River, NJ: Prentice Hall
250-word reply to classmate threads. The reply requires a minimum of 1 properly formatted citation. Each reply must be completed by you, the individual student. Additionally, each thread and reply must reflect a solid Christian worldview through the use of at least 1 Holy Bible reference.
Responding to a classmate’s post requires both the addition of new ideas and analysis. A particular point made by the classmate must be addressed and built upon by your analysis in order
to move the conversation forward
. Thus, the response post is a rigorous assignment that requires you to build upon initial posts to develop deeper and more thorough discussion of the ideas introduced in the initial posts. As such, reply posts that merely affirm, restate, or unprofessionally quarrel with the previous post(s) and fail to make a valuable, substantive contribution to the discussion will receive appropriate point deductions.
.
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International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
A Case of Postmenopausal Pyometra Caused By Endometrial Tuberculosisiosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Asymptomatic urinary tract infection amongst some Students of Michael Okpara ...Premier Publishers
This work was carried out to determine Asymptomatic Urinary Tract Infection amongst some students of Michael Okpara University of Agriculture, Umudike and the sensitivity pattern of the isolates from urine. Using aseptic technique, midstream urine were collected from sixty (60) students, urinalysis was carried out on the urine samples and was then cultured on CLED and MacConkey agar using pour plate method. Growth was observed in 26 (87%) of the sample while there was no growth in 4 (13%) of the sample. Out of the 26 (87%) samples with growth, 14 (47%) had significant bacteria growth while 12 (40%) had no significant growth. Incidence of asymptomatic bacteriuria was higher in females 8 (57%) than males 6 (38%). The organisms isolated were Escherichia coli, Klebsiella species, Staphylococcus saprophyticus, Staphylococcus aureus, Enterococcus faecalis, Proteus species, and Pseudomonas aeruginosa. All the Gram positive isolates were sensitive to Gentamycin and all resistance to Cefuroxime, Ceftazidime, Ceftriaxone, Cloxacillin. The Gram negative isolates were mostly sensitive to Nitrofurantoin, Gentamycin and Ofloxacin. Therefore, these drugs could be considered as the first line of drug for the treatment of asymptomatic urinary tract infection.
Ascaris lumbricoides and other Gastrointestinal Helminthic Parasites among Qe...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
FOURNIER’S GANGRENE: REVIEW OF 57 CASES IN TERTIARY INSTITUTIONAnil Haripriya
Fournier’s gangrene which is a rapidly progressive, fulminant polymicrobial synergistic infection of the perineum and genitals is now changing its pattern. Both genders can be affected and the mortality is still high (around10%). The clinical presentation in many patients in early stage may not be prominent. Thus rapid and accurate diagnosis is must for prompt treatment. Extensive surgical debridement and broad spectrum intravenous antibiotic remains the mainstay of treatment in order to reduce the morbidity and mortality.
Clinical Manifestations of Plasmodium bergheiANKA Infection in Juvenile Mice:...AI Publications
Malaria is an important health and development challenge in Africa, Animalmodels most particularly mice, have long been employedto study malaria pathogenesis. Clinical manifestations due to Plasmodium bergheiANKA infection in juvenile mice as a model for understanding the complications ofcongenital malaria in neonates.Forty-five juvenile mice (5-7 days old) were acquired from University College Hospital, Ibadan and injected with 2 x 107 (0.2ml) Plasmodium berghei ANKA parasitized red blood cells (PRBCs). Mice were transported to the study site, kept in well ventilated cages and fed daily with a balanced ration. Every day after post-P. berghei infection, mice were monitored for mortality. Clinical manifestations ofexperimental cerebral malaria (ECM) was assessed and confirmed if at leastruffled fur, hunching, wobbly gait, limb paralysis, convulsions, or coma was observed. Each sign was given a score of 1. Animals with scores ≥4 were considered to have severe ECM.20 (44%) micewerelost due to natural cause (i.e. stress) at day 2 of the experiment. Between day 4 and 9, 25 (56%) of the studymice presented clinical signs of ECM which includes; ruffled fur 25(100%), hunching 21 (84%), wobbly gait 17 (68%), limb paralysis 20 (80%), convulsions 25 (100%) and subsequently died. Survival rate and severity of ECM in the mice differs, 22 (88.0%) had severe ECM and 3(12.0%) had mild ECM.This study has shown that parasite establishment and malaria complications can manifest as early as 4 days’postP. berghei infection in 5-7 days old mice.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
The prevalence of Escherichia coli Causing Urinary Tract Infections in Aminu ...Premier Publishers
Urinary Tract infections (UTIs) are one of the most common causes of hospital visit worldwide. The study intends to find the prevalence of Escherichia coli in Urinary tract infection cases in Aminu Kano Teaching Hospital (AKTH), Kano. A comprehensive study was conducted on E coli for its prevalence in urine samples, of queried cases of urinary tract infection in patients attending Aminu Kano Teaching Hospital (AKTH), Kano. Two hundred and fourteen urine specimens comprising of 123(57.5%) females and 91(42.5%) males of all age group were screened for bacteria. Of the 214 samples, 68 representing (31.80%) were culture positive with E. coli having 32(47.06%), Klebsiella spp 18(26.47%), Staphylococcus spp 10(14.70%), Proteus spp 6(8.82%) and Pseudomonas spp 2(2.94%). Out of the 32(47.6%) of E. coli isolated, 22 were from female and 10 were from males, the highest prevalence occurred within the age range of 21-30 with females taking the lead. Antibiogram of the isolated E. coli showed a markedly good sensitivity of ofloxacin (96.87%), nitrofurantoin and nalidixic acid (93.70%), colistin phosphate (90.62%) and gentamicin (68.75%). E. coli showed the least sensitivity to ampicillin (18.75%). The frequency of E. coli obtained suggests its high prevalence. And this can be reduced using Flouroquinolones as exhibited by the susceptibility profile in this study.
Similar to 1ScIeNtIFIc REPORTS (2018) 81250 DOI10.1038s41598-018.docx (20)
COLLEGE
PHYSICS LAB REPORT
STUDENTS NAME
ANALYSIS OF A BUBBLE CHAMBER PICTURE
SUPERVISED BY:
19/05/2020
1. Introduction
A bubble chamber is a vessel filled with a superheated transparent liquid (most often liquid hydrogen) used to detect electrically charged particles moving through it. It was invented in 1952 by Donald A. Glaser, for which he was awarded the 1960 Nobel Prize in Physics.
A convenient way to study the properties of the fundamental subatomic particles is through observation of their bubble trails, or tracks, in a bubble chamber. Using measurements made directly on a bubble chamber photograph, we can often identify the particles from their tracks and calculate their masses and other properties. In a typical experiment, a beam of a particular type of particle is sent from an accelerator into a bubble chamber, which is a large liquid-filled vessel. To simplify the analysis of the data, the liquid used is often hydrogen, the simplest element. The use of liquid hydrogen, while it simplifies the analysis, complicates the experiment itself, since hydrogen, a gas at room temperature, liquefies only when cooled to -246◦C. For charged particles to leave tracks in passing through the chamber, the liquid must be in a “super-heated” state, in which the slightest disturbance causes boiling to occur. In practice, this is accomplished by expanding the vapor above the liquid with a piston a few thousandths of a second before the particles enter the chamber.
2. Methods
2.1 Materials needed:
1. student worksheet per student
2. Ruler
3. Scissors
4. Glue stick
5. Pocket calculator
2.2 Procedures
2.2.1 Calculation of the X Particle’s Mass.
Make measurements on each of the photographs. In particular, for each of the circled events measure these four quantities:
· `Σ - The length of the Σ track,
· θ - the angle between the Σ− and π− track,
· s - the sagitta of the π− track,
· `π - The chord length of the π− track.
Your values for the event should be close to those given in the sample input. Run the program using each set of measurements, and tabulate the computed X0 mass from each event. Compute an average of the calculated masses and find the average deviation, expressing your result as Mx ±∆Mx.
Compare your final result with some known neutral particles listed below and identify the X0 particle based on this comparison.
Particlemass (in MeV/c2)
π0 135
K0 498
n 940
Λ0 1116
Σ0 1192
Ξ0 1315
2.2.2 Determination of the Angle θ.
The angle θ between the π− and Σ− momentum vectors can be determined by drawing tangents to the π− and Σ− tracks at the point of the Σ− decay.
We can then measure the angle between the tangents using a protractor. We can show.
Collins did not understand the events that led to the reasoning .docxvickeryr87
Collins did not understand the events that led to the reasoning why and actions how Wells Fargo conveyed victories, achievements, and accomplishments. Therefore, Collins was incorrect. Wells Fargo as a company was able to achieve earning because of their successful strategies in profit revenue or turnover for each employee. The Good to Great article brought up that many people were surprised when finding out what made companies successful especially in relation to the leadership (Collins, 2001).
Our text states that it is difficult for companies to find good ethical leaders because they tend to be more expensive (Meese, Ortmeier, 2001). Unethical leadership is sure to bring the downfall of any business; it is impossible to have a successful business if there are unethical practices, the employees are also bound to begin these practices making it difficult for the company to hold their end of their promises to their customers or members.
"He who oppresses the poor to make more for himself or who gives to the rich, will only come to poverty" (Proverbs 22:16).
Collins, J. (2001). Good to great: Why some companies make the leap and others don't. New York, NY: Harper Collins.
Mees, E., Ortmeier, P. (2001). Leadership, ethics, and policing:Challenges for the 21st century. Upper Saddle River, NJ: Prentice Hall
250-word reply to classmate threads. The reply requires a minimum of 1 properly formatted citation. Each reply must be completed by you, the individual student. Additionally, each thread and reply must reflect a solid Christian worldview through the use of at least 1 Holy Bible reference.
Responding to a classmate’s post requires both the addition of new ideas and analysis. A particular point made by the classmate must be addressed and built upon by your analysis in order
to move the conversation forward
. Thus, the response post is a rigorous assignment that requires you to build upon initial posts to develop deeper and more thorough discussion of the ideas introduced in the initial posts. As such, reply posts that merely affirm, restate, or unprofessionally quarrel with the previous post(s) and fail to make a valuable, substantive contribution to the discussion will receive appropriate point deductions.
.
Define discrimination, victimization and affirmative actions; .docxvickeryr87
Define discrimination, victimization and affirmative actions; and consider the ethical and legal aspects of each
Consider problems with sexual harassment and workplace drugs, and consider the ethical implication of company policies directed at each
Reference
American Public University System. (n.d.). MGMT200: Lesson 5: The Corporation and External Stakeholders. Retrieved from https://apus.realizeithome.com/
Collins, D. (2012). Business Ethics. Hoboken, NJ: Wiley
Weiss, J. (2014).
Business Ethics: A Stakeholder & Issues Management Approach
. San Francisco, CA: Koehler.
.
Define data mining. Why are there many names and definitions for d.docxvickeryr87
Define data mining. Why are there many names and definitions for data mining
What are the main reasons for the recent popularity of data mining?
Discuss what an organization should consider before making a decision to purchase data mining software.
.
Define culture. How can culture be conceptionalizedDiscuss at l.docxvickeryr87
Define culture. How can culture be conceptionalized?
Discuss at least two of the variables that moderate differences between domestic and international HR practices.
Compare cross-cultural management studies, and list their advantages and disadvantages.
Discuss two HR activities in which a multinational firm must engage, which would not be required in a domestic environment?
200 words
In text citations
Dowling, P. J., Festing, M., & Engle, A. (2013).
(6th ed.). Boston, MA:
International human resource management Cengage
.
Define cultural relativism and how it is used by anthropologis.docxvickeryr87
Define cultural relativism and how it is used by anthropologists as an approach to cross-cultural research. Explain how cultural relativism contributes to anthropologists' efforts to counter ethnocentrism.
Describe an example of how your culture has ‘taught’ you to behave the way you do, or to believe what you believe. This could be an instance in which you were taught a lesson, or perhaps when you made a mistake and were corrected by an elder. Think about how such lessons shape your worldview, your ideas about what's "normal," and your values. Are these universal beliefs or are they culturally constructed?
I need the answer in the form of a speech draft cause I need to use it on presentation.!!!!!!!!!!!!
First question no less than 200 words, the second question no less than 150 words
.
Define cost control and provide several examples of how it affec.docxvickeryr87
Define cost control and provide several examples of how it affects managers at all levels within a healthcare organization. Compare and contrast Medicare and Medicaid in terms of eligibility, benefit packages, access to care, and other key dimensions. Discuss the various ways in which healthcare reform has affected and may affect the financial delivery of health care today and into the future.
.
Define corporate governance.Discuss the events that led up.docxvickeryr87
Define corporate governance.
Discuss the events that led up to the need for increased corporate governance.
Define business strategy. List five possible business strategies.
Discuss the purpose of the Capability Maturity Model.
Define auditing.
Define internal control and provide an example of a control.
Discuss the role Sarbanes/Oxley has played with respect to corporate governance.
Define IT governance.
List factors one should consider with respect to governing the cloud.
I need answers with references and intext citations.
.
Define communication in your own words. Identify and distinguish amo.docxvickeryr87
Define communication in your own words. Identify and distinguish among the five forms of communication and the persuasive speech ( please read content). I’m also including my two papers so you can compare. my art and music essay is my persuasive essay it’s also my best essay please read content.......
.
Define Civil Liberties. List 5 Civil Liberties. How do they differ.docxvickeryr87
Define Civil Liberties. List 5 Civil Liberties. How do they differ from Civil Rights? Which groups have challenged government and individuals to obtain Civil Rights, and how? Provide at least three examples of civil rights legislation that were a result of civil rights movements.
.
Define civilization. Do we really need it in order to survive and pr.docxvickeryr87
Define civilization. Do we really need it in order to survive and prosper on this planet? The real question is whether we can live above subsistence without the technological innovations that cities generate. Moreover, the question is not as frivolous as it may seem at first glance. Many cultures throughout human history consciously decided not to take the route toward nucleated settlements and the resultant consequences. The Celts represent one example from the time line of our course of a people who preferred not to have cities. The Avars, a Turkic-speaking people from Central Asia, were pastoral nomads who acquired cities during the course of their conquests, yet they “used” cities in ways that would strike average city dwellers as most strange.
Section 2.1 of your textbook,
World History: The Human Experience to 1500
, presents nine characteristics commonly associated with civilizations. Choose one society from the
Week One Civilization Tour
, which includes Mesopotamian, Olmec, Egyptian, Harappan, and Ancient Chinese societies. Based on at least two of the characteristics listed in Section 2.1, explain why your chosen society can be considered a civilization. Provide specific examples drawn from this week’s learning materials to support your claims. Cite all sources according to APA style as outlined in the
Ashford Writing Center
, using both in-text citations and providing full references at the bottom of your post.
Your initial post should be a minimum of 300 words. Respond to two colleagues in a substantive post of at least 150 words.
.
Define case management and care management and compare the dif.docxvickeryr87
Define case management and care management and compare the differences.
Mention and discuss the case management concepts into the clinical practice of community health nursing.
Give the definition of family, mention, and discuss the different types of families, mention, and discuss the model of care for families.
Describe strategies for moving from intervention at the family level to intervention at the aggregate level
.
Define Bureaucracy. Government at all levels has grown enormously, .docxvickeryr87
Define Bureaucracy. Government at all levels has grown enormously, how is the federal bureaucracy organized. (give examples of the 4 levels) . How has bureaucracy grown over time and why? Why might independent regulatory agencies be considered the fourth branch of government? What are iron triangles and issue networks and how do they affect policymaking? Describe recent attempts to reduce bureaucratic inefficiency and improve performance?
.
Define and explain how the Twitter search function works to search f.docxvickeryr87
Define and explain how the Twitter search function works to search for specific words in any Twitter post.
In developing your initial response, be sure to draw from, explore, and cite credible reference materials, including
at least one scholarly peer-reviewed reference
. In responding to your classmates’ posts, you are encouraged to examine their opinions, offering supporting and/or opposing views.
.
Define and relate these different terminologies and Information Gove.docxvickeryr87
Define and relate these different terminologies and Information Governance (IG), Data Governance (DG) and Information Technology Governance (ITG), E-Discovery, Digital Assets; how do you see IT Governance evolving in the next 5 years? What framework will you adopt to in implementing the IT Governance Initiative, and demonstrate its applicability.
.
Define and provide examples of-Basic probability- Bayes the.docxvickeryr87
Define and provide examples of:
-Basic probability
- Bayes theorem
- Multiplication rule
- Addition rule
-Binomial distribution
-Poisson distribution
-Normal distribution
-Sample distribution
-Central limit theorem
Use Apa Style, cites, references and avoid plagiarism.
.
Define and discuss the phrase Manifest Destiny. Explain how this b.docxvickeryr87
Define and discuss the phrase "Manifest Destiny." Explain how this belief came to divide the nation.
Your response should be at least 200 words in length. You are required to use at least your textbook as source material for your response. All sources used, including the textbook, must be referenced; paraphrased and quoted material must have accompanying citations.
.
Define and discuss the differences between vision and mission stat.docxvickeryr87
Define and discuss the differences between vision and mission statements.
Your response should be at least 200 words in length. All sources used, including the textbook, must be referenced; paraphrased and quoted material must have accompanying citations.
David, F. (2011). 1.
Strategic management: concepts & cases
(Custom Edition ed., pp. 11-13). New York: McGraw-Hill Irwin.
No Wiki, Dictionary.com or Plagiarism
.
Define and discuss the four types of innovation. How might these.docxvickeryr87
Define and discuss the four types of innovation. How might these innovations relate to the industry life cycle?
business innovation is:
Discuss the ‘make versus buy decision firms often are forced to make (Define terms)
What is vertical integration? Discuss to include benefit, risk, possible alternative, relationship to company value chain.
.
Define and discuss the data wiping process.Discuss how a cloud.docxvickeryr87
Define and discuss the data wiping process.
Discuss how a cloud-based solution provider may reduce the risk of a DDoS attack.
Define and discuss hyperjacking attacks.
Define and discuss guest-hopping attacks.
Paper requirements
:
Minimum
1200
words (excluding title page, table of contents, abstract, and references pages)
Minimum of four (4) references
Format your paper consistent with APA guidelines
When submitting the assignment, please ensure you are submitting as an attached
MS Word document
.
.
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
for beginners, providing thorough training in areas such as SEO, digital communication marketing, and PPC training in Noida. After finishing the program, students receive the certifications recognised by top different universitie, setting a strong foundation for a successful career in digital marketing.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
1. 1ScIeNtIFIc REPORTS | (2018) 8:1250 | DOI:10.1038/s41598-
018-19638-x
www.nature.com/scientificreports
Histology, immunohistochemistry,
and in situ hybridization reveal
overlooked Ebola virus target
tissues in the Ebola virus disease
guinea pig model
Timothy K. Cooper1, Louis Huzella 1, Joshua C. Johnson 1,
Oscar Rojas1, Sri Yellayi1,3,
Mei G. Sun2, Sina Bavari2, Amanda Bonilla1, Randy Hart1,
Peter B. Jahrling 1, Jens H. Kuhn 1
& Xiankun Zeng 2
Survivors of Ebola virus infection may become subclinically
infected, but whether animal models
recapitulate this complication is unclear. Using histology in
combination with immunohistochemistry
and in situ hybridization in a retrospective review of a guinea
pig confirmation-of-virulence study, we
demonstrate for the first time Ebola virus infection in hepatic
oval cells, the endocardium and stroma of
the atrioventricular valves and chordae tendinae, satellite cells
of peripheral ganglia, neurofibroblasts
and Schwann cells of peripheral nerves and ganglia, smooth
muscle cells of the uterine myometrium
and vaginal wall, acini of the parotid salivary glands, thyroid
follicular cells, adrenal medullary cells,
pancreatic islet cells, endometrial glandular and surface
epithelium, and the epithelium of the vagina,
2. penis and, prepuce. These findings indicate that standard animal
models for Ebola virus disease are not
as well-described as previously thought and may serve as a
stepping stone for future identification of
potential sites of virus persistence.
Ebola virus disease (EVD) is a severe and frequently lethal
affliction of humans caused by infection with any
of three members of the mononegavirus family Filoviridae:
Bundibugyo virus (BDBV), Ebola virus (EBOV),
and Sudan virus (SUDV). A fourth virus, Taï Forest virus
(TAFV), has thus far caused only a single reported
human infection, which was nonlethal1. EVD is an exotic
disease with case numbers rarely surpassing the lower
hundreds1; however, from 2013–2016, EBOV caused an EVD
outbreak in Western Africa encompassing 28,616
infections and 11,310 deaths in Guinea, Liberia, and Sierra
Leone2. Long term sequelae in individual survivors of
acute EVD and the similar Marburg virus disease (MVD) and
filovirus persistence followed by disease relapse or
sexual transmission had been reported before this outbreak3–8.
However, observations during and following the
Western African EVD outbreak suggest that sequelae and
filovirus persistence may be common events9. Reported
sequelae include arthralgia, cardiac valvulopathy, parotid gland
inflammation, peripheral paresthesia or dyses-
thesia, and gastrointestinal motility disorders10–14. Semen may
contain detectable EBOV RNA for more than 500
days following recovery, and EBOV RNA has been detected in
breast milk of a subclinically infected mother15,16.
Replicating EBOV has been isolated from the cerebrospinal
fluid of an EVD survivor suffering a disease relapse
and from the aqueous humor of the eye of another
survivor17,18. Sexual EBOV transmission from EVD survivors
to partners months after infection also has been
documented19,20.
3. Only 26 EVD outbreaks (16 of them due to EBOV infection)
have been recorded since the initial discovery of
EVD-causing viruses in 1976, and excluding the Western
African outbreak, no EVD outbreak has encompassed
more than 500 cases1. Because the natural host reservoirs of
EVD-causing filoviruses remain to be identified,
1Integrated Research Facility at Fort Detrick, National Institute
of Allergy and Infectious Diseases, National Institutes
of Health, Fort Detrick, Frederick, Maryland, USA. 2United
States Army Medical Research Institute of Infectious
Diseases, Fort Detrick, Frederick, Maryland, USA. 3Present
address: Path-2-Gene, LLC, Harrisburg, PA, USA. Timothy
K. Cooper and Xiankun Zeng contributed equally to this work.
Correspondence and requests for materials should be
addressed to T.K.C. (email: [email protected]) or X.Z. (email:
[email protected])
Received: 9 November 2017
Accepted: 2 January 2018
Published: xx xx xxxx
OPEN
http://orcid.org/0000-0003-2913-489X
http://orcid.org/0000-0002-5677-3841
http://orcid.org/0000-0002-9775-3724
http://orcid.org/0000-0002-7800-6045
http://orcid.org/0000-0003-3526-8755
mailto:[email protected]
mailto:[email protected]
4. www.nature.com/scientificreports/
2ScIeNtIFIc REPORTS | (2018) 8:1250 | DOI:10.1038/s41598-
018-19638-x
the periodic introductions of these viruses into the human
population are unpredictable and unpreventable. In
conjunction with the high case-fatality rate of EVD1,
performance of meaningful clinical studies is challenging.
Therefore, characterization of EVD pathogenesis and primary
development and evaluation of medical counter-
measures largely relies on experimental infections of animal
models. Predominantly, these models include non-
human primates (crab-eating, aka cynomolgus, macaques
[Macaca fascicularis] and rhesus monkeys [Macaca
mulatta]), and, after virus adaptation, rodents (laboratory mice,
guinea pigs [Cavia porcellus], and Syrian hamsters
[Mesocricetus auratus])21,22. Recently, domestic ferrets
(Mustela putorius furo) have been identified as novel EVD
animal models not requiring virus adaptation23–25. However,
with the exception of one study demonstrating EBOV
persistence in apparently healthy rhesus monkey survivors of
EBOV-induced disease26, development of sequelae
and/or viral persistence, and demonstration of sexual
transmission, remains to be recapitulated in animal models.
Based on in vitro results of studies of EBOV persistence in cell
culture27, we speculate that EBOV may establish
in vivo persistent infections in any entry-permissive cell type,
rather than only in immunoprivileged sites such
as brain, eyes, and testes. Before this speculation can be tested,
validating the reported tissue and cell tropism of
EBOV in animal models is imperative. We chose the guinea pig
model, as the model is considered to be more
stringent than the laboratory mouse model and less ethically,
financially, and logistically burdensome than non-
5. human primates models21,22. The guinea pig model is
considered well-characterized and is actively used for trans-
mission and medical countermeasure evaluation studies. EBOV
infection has been previously demonstrated in a
wide variety of guinea pig tissues and cell types, including cells
of the monocyte or macrophage system, hepato-
cytes, fibroblastic reticular cells, interstitial fibroblasts, adrenal
cortical cells, ovarian thecal cells, endothelial cells,
endometrial stromal cells, and cells of the urothelium28–39. In
a retrospective histologic review of tissues from
EBOV-infected guinea pigs, in combination with
immunohistochemistry and in situ hybridization, we demon-
strate that EBOV infection of several tissues and numerous cell
types has been overlooked.
Results
All 30 guinea pigs intraperitoneally infected with 1 of 3 EBOV
doses (n = 10/group) developed illness. Seven
guinea pigs (3 in 10-PFU group, 2 in 100-PFU group, and 2 in
the 1,000-PFU group) succumbed to EVD and
were not necropsied due to autolysis. The remaining 23 guinea
pigs (7 in the 10-PFU group, 8 in the 100-PFU
group, and 8 in the 1,000-PFU group) were necropsied, and
specific tissues were collected for histology (see
Supplemental Table 1 for animal-by-animal breakdown of sex,
inoculation dose, day of euthanasia, and relevant
pathology findings). All 23 guinea pigs examined histologically
had extensive hepatocellular and lymphoid necro-
sis consistent with previous descriptions of guinea pig-adapted
Ebola virus infection in this model30,31. IHC and
ISH staining confirmed hepatocytes, monocytes and
macrophages as major targets of EBOV infection.
Liver. The majority of findings were consistent with previous
reports30,31, including widespread hepatocyte and
Kupffer cell infection and necrosis, with variable dystrophic
6. mineralization. In contrast to the rhesus monkey model
of EVD, viral intracytoplasmic inclusion bodies (ICIB) were
large and numerous. Livers were collected from 6/7
guinea pigs in the 10-PFU, 8/8 in the 100-PFU, and 8/8 in the
1,000-PFU group (Supplemental Table 1). In 7 of 22
livers examined (samples from 1 guinea pig were not available),
multifocal minimal- to-moderate oval cell hyper-
plasia originated in the biliary ductules and extended into
lobules. Small basophilic cells with a high nucleus:cy-
toplasm ratio formed parallel streams and duct-like structures,
consistent with International Harmonization of
Nomenclature and Diagnostic criteria (INHAND)40. Viral ICIB
were frequently observed within oval cells (Fig. 1a)
with IHC and ISH positive signals (Fig. 1b,c). Oval cells were
present in animals from all three dosage groups, with
the occurrence of such cells loosely correlating with the
infectious inoculum: 1 of 6 guinea pigs in the 10-PFU group;
1 of 8 in the 100-PFU group; and 5 of 8 in the 1000-PFU group.
Oval cells were only present in animals necropsied
at d 8 or 9 post-inoculation. No viral infection of biliary
ductules was present as judged by histology, IHC, or ISH.
Heart. Cross-sections through one or both ventricles were
collected from 22 of 23 guinea pigs (samples from
1 guinea pig were not available), with variable inclusion of
atrioventricular valves and associated chordae tendi-
nae; semilunar valves were not sampled. In all 3 examined
tricuspid valves (one each from the 10-, 100-, and
1,000-PFU groups) and both mitral valves (one each from the
100- and 1000-PFU groups), viral ICIB were pres-
ent in the stromal spindled fibroblasts of the spongiosa and
surface endocardium of atrialis of the atrioventricular
valves (Fig. 1d). Sometimes viral ICIBs were associated with
infiltrating macrophages and apoptotic and necrotic
cellular debris (Fig. 1d). EBOV infection of the fibroblasts was
confirmed by IHC and ISH (Fig. 1e,f ). Viral ICIBs
7. were also present in the stromal cells of the chordae tendinae of
the tricuspid valve in 8 of 11 examined guinea
pigs and of the chordae tendinae of the mitral valve of 16 of 17
examined guinea pigs. In three of 22 guinea pigs,
very rare individual cardiomyocytes were noted that contained
ICIB, with confirmation by IHC.
Nerves and ganglia. From samples available from 7 guinea pigs,
the paradrenal or mesenteric ganglia and
nerves were collected with adrenals or mesenteric lymph nodes,
respectively. Multifocal mild to rarely moderate
histiocytic inflammation (ganglioneuritis) was noted in 5 of 7
animals, with mild ganglion cell degeneration or
loss and satellite cell hyperplasia (nodules of Nageotte). Viral
ICIB were present within infiltrating macrophages,
and occasional satellite cells, neurofibroblasts, and Schwann
cells (Fig. 2a), with EBOV-positive labeling by IHC
and ISH (Fig. 2b,c). No evidence of neuronal infection was
observed in any sample. Multiple myelinated medium
and large mesenteric nerves and other peripheral nerves were
characterized by rare ICIB within neurofibroblasts,
Schwann cells, and infiltrating macrophages (Fig. 2d). Infected
cells, endothelial cells, and intravascular plasma
were labeled by IHC and ISH (Fig. 2e,f ). In a single guinea pig,
multiple digestion chambers consisting of segmen-
tal expansion of the axon sheath by lipid laden macrophages
were present in a single large axillary/brachial nerve
in the absence of ICIB. Unfortunately, the nerve was lost on re-
sectioning.
www.nature.com/scientificreports/
3ScIeNtIFIc REPORTS | (2018) 8:1250 | DOI:10.1038/s41598-
018-19638-x
8. Genital tract. Of the 12 EBOV-infected female guinea pigs for
which slides were available, uterine bod-
ies were collected from 3 animals, uterine cervices and vaginas
from 3 animals, and vaginas from 2 animals.
In all specimens examined except one vagina, rare-to-frequent
viral ICIB were present within smooth muscle
cells, with scant apoptosis, necrosis, or histiocytic inflammation
(Fig. 3a–c). In two sections of uterus and two
sections of vagina, immunolabeling of rare small clusters of
surface and glandular epithelial cells was positive
(Fig. 4a,b). Consistent with previous reports, endometrial
stromal cells and interstitial fibroblasts, macrophages
and endothelial cells contained ICIB and stained positively for
EBOV by IHC and ISH30,31.
The penis and prepuce was collected from a single male guinea
pig (post-inoculation day 9, 100-PFU group).
Several small clusters of spinous layer epithelial cells in the
prepuce exhibited cytoplasmic vacuolation and
Figure 1. EBOV infection of the liver and heart. (a) Oval cell
infection of the liver. Parallel streams of
basophilic oval cells form duct-like structures adjacent to a
portal triad, with frequent large eosinophilic
toamphiphilic intracytoplasmic inclusion bodies (arrows, H&E).
(b) Proliferating oval cells, hepatocytes,
Kupffer cells, and intravascular monocytes (EBOV VP40 IHC:
DAB chromogen and hematoxylin). (c) Oval
cells, hepatocytes, and Kupffer cells (EBOV NP ISH: fast red
chromogen and hematoxylin). (d) Valvular
endocarditis of tricuspid valve cusp. Degeneration and necrosis
of surface (atrialis) endocardial cells and
stromal cells of the spongiosa, with infiltrating macrophages
and frequent large eosinophilic to amphiphilic
intracytoplasmic inclusion bodies (arrows, H&E). (e)
9. Endocardial cells, stromal cells, and macrophages (EBOV
VP40 IHC: DAB chromogen and hematoxylin). (f) Endocardial
cells, stromal cells, and macrophages (EBOV
NP ISH: fast red chromogen and hematoxylin).
www.nature.com/scientificreports/
4ScIeNtIFIc REPORTS | (2018) 8:1250 | DOI:10.1038/s41598-
018-19638-x
contained viral ICIB (Fig. 4c). Scattered apoptosis and necrosis
of adjacent cells were noted. Discrete vertical col-
umns of EBOV-positive cells (IHC) were present in the non-
haired skin of both the penis and prepuce (Fig. 4d–f ).
Salivary glands. Salivary glands were collected incidentally
with the mandibular lymph nodes from some
animals. In most salivary glands, very low numbers of
macrophages and fibroblasts containing ICIB were
present in the interstitium surrounding the acini and ducts. In 8
of 10 parotid glands examined, multifocal
mild-to-moderate acinar cell necrosis with histiocytic
inflammation was observed (Fig. 5a). Immunostaining of
3 glands showed abundant EBOV antigen localized to these
necrotic foci and in viable and degenerate acinar cells
Figure 2. EBOV infection of paradrenal ganglia and peripheral
medium myelinated nerves. (a) Low numbers
of macrophages infiltrate between ganglion cells in the
paradrenal ganglion. Viral intracytoplasmic inclusion
bodies (arrows) are present within satellite cells,
neurofibroblasts, Schwann cells, and macrophages (H&E).
(b) Satellite cells, neurofibroblasts, Schwann cells, and
macrophages, and intravascular plasma (EBOV VP40
10. IHC: DAB chromogen and hematoxylin). (c) Satellite cells,
neurofibroblasts, Schwann cells, and macrophages
in the ganglion (EBOV NP ISH: fast red chromogen and
hematoxylin). (d) Viral intracytoplasmic inclusion
bodies (arrows) are present within neurofibroblasts and
Schwann cells within a nerve, with low numbers of
perivascular infiltrating macrophages (bottom) (H&E). (e) Rare
infected Schwann cells and neurofibroblasts
within a myelinated nerve (EBOV VP40 IHC: DAB chromogen
and hematoxylin). (f) Low numbers of Schwann
cells, neurofibroblasts, and infiltrating macrophages within a
myelinated nerve (EBOV NP ISH: fast red
chromogen and hematoxylin).
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(Fig. 5b). ISH results confirmed these IHC findings (Fig. 5c).
Rare scattered acinar cell necrosis without ICIB was
noted in 1 of 8 submandibular glands examined. Rare ductal cell
necrosis in the absence of ICIB was observed in
2 of 6 sublingual glands available for review.
Figure 3. EBOV infection of the uterus. (a) Numerous large
eosinophilic to amphiphilic intracytoplasmic
inclusion bodies (arrows) within smooth myocytes, with
scattered apoptosis and necrosis (H&E). (b) Smooth
myocytes and endometrial stromal cells (EBOV VP40 IHC:
DAB chromogen and hematoxylin). (c) Myometrial
smooth muscle cells (EBOV NP ISH: fast red chromogen and
hematoxylin).
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Figure 4. EBOV infection of the vagina, penis, and prepuce. (a)
Rare small clusters of infected metestrus
epithelial cells in the vagina (EBOV VP40 IHC: DAB
chromogen and hematoxylin). (b) Endometrial gland
epithelial cells, as well as stromal cells and macrophages, of the
uterine body (EBOV VP40 IHC: DAB
chromogen and hematoxylin). (c–f) Penis and prepuce. (c)
Scattered small clusters of keratinocytes within the
stratified squamous cornifying epithelium with large viral
intracytoplasmic inclusions (arrows). (d) In addition
to stromal fibroblasts and macrophages, clusters of epithelial
cells covering the penis and prepuce (EBOV VP40
IHC: DAB chromogen and hematoxylin). (e) Higher
magnification of subfigure. (d,f) Clusters of infected
epithelial cells. (EBOV GP IHC: DAB chromogen and
hematoxylin).
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Thyroid. Thyroid tissues were collected incidentally with the
trachea and esophagus from 4 of 23 guinea pigs
examined. In all glands, very low numbers of macrophages and
fibroblasts containing ICIB were present in the
interstitium surrounding the follicles. In 3 of the 4 thyroid
12. tissues available, multifocal mild-to-moderate follicu-
lar cell degeneration and necrosis were noted with low numbers
of infiltrating viable and degenerate macrophages
within the follicles (Fig. 5d). Both macrophages and follicular
cells contained ICIB and were immunopositive,
with occasional granular staining of the colloidal material
similar to the staining of plasma within blood vessels
(Fig. 5e). Macrophages, interstitial fibroblasts and follicular
epithelial cells were also positive for EBOV RNA by
ISH (Fig. 5f ).
Figure 5. EBOV infection of the salivary glands and thyroid. (a)
Necrotizing adenitis of parotid salivary gland.
Focal acinar cell necrosis with rare large eosinophilic to
amphiphilic intracytoplasmic inclusion bodies (arrows,
H&E). (b) Infiltrating macrophages, interacinar fibroblasts, and
viable and degenerate acinar cells (EBOV VP40
IHC: DAB chromogen and hematoxylin). (c) Infected acinar
cells, macrophages and fibroblasts (EBOV NP
ISH: fast red chromogen and hematoxylin). (d) Degeneration,
sloughing, and necrosis of follicular epithelium,
with rare large eosinophilic to amphiphilic intracytoplasmic
inclusion bodies (arrows) in follicular cells and
infiltrating macrophages (H&E). (e) Infected follicular cells,
intra-follicular and interstitial macrophages,
and interstitial stromal cells (EBOV VP40 IHC: DAB
chromogen and hematoxylin). (f) Infected follicular
epithelium and interstitial cells (EBOV NP ISH: fast red
chromogen and hematoxylin).
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13. Adrenal Glands. Consistent with previous reports on the guinea
pig model, adrenal cortical cells and sinu-
soidal macrophages were frequently infected30,31,33,41. Low
numbers of chromaffin cells (less than 5%) with viral
ICIB were present in the adrenal medullas from 5 of the 18
guinea pigs from which they were collected (Fig. 6a–c).
Glassy eosinophilic “hyaline globules” within the cytoplasm of
medullary cells were noted more frequently than
viral ICIB42.
Pancreas. Of the 23 guinea pigs available, 6 pancreata were
examined. Very low numbers of macrophages and
fibroblasts containing ICIB were noted in the interstitium
surrounding acini and ducts, consistent with previous
reports30. The presence of ICIB and positive EBOV IHC and
ISH signals within acinar cells were also expected. In
5 of the 6 pancreata, low numbers of endocrine cells within the
islets of Langerhans contained ICIB and stained
positively by IHC and ISH, frequently occurring in small
clusters (Fig. 6d,e). Islets were not uniformly affected,
with some islets containing no infected cells while rare islets
had up to 20% EBOV infection.
Figure 6. EBOV infection of adrenal medulla and pancreas. (a)
Occasional large viral inclusions (arrows)
within chromaffin cells (H&E). (b) Chromaffin cells (EBOV
VP40 IHC: DAB chromogen and hematoxylin).
(c) EBOV-infected chromaffin cells (EBOV NP ISH: fast red
chromogen and hematoxylin). (d and e) EBOV-
infected endocrine cells of the Islets of Langerhans, exocrine
acinar cells, and interstitial fibroblasts and
macrophages (d: EBOV VP40 IHC: DAB chromogen and
hematoxylin, e: EBOV NP ISH: fast red chromogen
and hematoxylin).
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Discussion
The gross and histologic lesions of experimental EBOV-induced
disease in nonhuman primate and rodent mod-
els are considered largely identical with that of natural human
infections30,31,43. However, this presumed identity
may be incorrect because, for instance, only very few autopsies
of EVD victims have been performed43. Likewise,
despite the long history of using guinea pigs in filovirus
research28–39, many manuscripts do not include histology
or are limited to the evaluation of livers and spleens. Only a
handful of studies have been performed to thoroughly
characterize EBOV cell tropism in the guinea pig using
histology, ultrastructural evaluation and/or IHC, and
almost none have utilized ISH to detect viral
genome30,31,33,41. In our study, we discovered that EBOV
infects and
likely replicates in numerous tissues and cell types previously
not known to be involved in the pathogenesis in the
guinea pig model.
Although not previously described in filovirus infection models,
the finding of oval cell hyperplasia was not
unexpected because of the extensive hepatocellular necrosis
subsequent to EBOV infection. Oval cells, derived
from the terminal ductule epithelial cells of the canal of Hering,
are bipotential progenitor cells that may dif-
ferentiate to biliary cells or hepatocytes40. Oval cells may be
induced with any liver injury, including toxins and
infections. However, we were surprised by the presence of
15. EBOV RNA and antigen in oval cells, which could
potentially affect both EBOV clearance and hepatic
parenchymal regeneration in EVD survivors.
EBOV infection and inflammation of the atrioventricular valves
and chordae tendinae of the heart was also
unexpected, but could possibly explain valvulopathy in
survivors, either as a result of inflammation and scarring
or of autoimmune disease. Similarly, infection of the chordae
tendinae could predispose to rupture of the tendi-
nae, resulting in atrioventricular valve prolapse. Infection of the
atrial myocardium and vascular endothelium
and tunica media has been previously described, but whether
valves were included in previous examinations is
unclear30. Demonstration of abundant viral antigen in the
endocardium (NOS) has been reported in fatal acute
infections in humans43.
EBOV infection of the supporting cells of the peripheral ganglia
and nerves, with attendant inflammation, is
intriguing, as the nature of neurologic impairment in EVD
survivors is hotly debated17,26,44,45. Damage to these
neural supporting cells, with a pro-inflammatory resolution,
could conceivably lead to long-term nerve or gan-
glion damage, manifesting as altered sensory inputs or
dysautonomia. Therefore, prospective studies of a survivor
model of EVD, once established, should include thorough
sampling of the peripheral nervous system to test this
hypothesis.
Infection of ovarian thecal cells and stromal cells and
macrophages of the ovary, oviduct, and uterine endome-
trium has been described previously in the guinea pig model of
EVD and MVD30,46. Infection of smooth myocytes
of the vascular tunica media has also been described30. Thus,
the finding of smooth myocyte infection in the
16. guinea pig uterus, although novel, was not entirely surprising.
Although a minor feature, epithelial cell infection
of the endometrium, vagina and penis/prepuce is highly
interesting vis-à-vis potential sexual transmission of
virus during acute subclinical or prodromal infection4,19,20.
Other mucosal epithelia such as those of the tongue
and esophagus have been previously demonstrated by others to
be infected30,32.
The high incidence of EBOV infection and inflammation of the
parotid salivary gland is significant, as saliva
is known to contain the virus in acutely infected and recently
convalescent individuals, and inflammation of the
parotid gland is a documented sequela of disease in survivors47.
The preference of EBOV for the parotid gland
over the submandibular and sublingual glands may explain
negative findings in a previous study, in which the
specific salivary gland(s) examined was/were not reported31.
Preferential infection of the parotid salivary gland
has been described for other viruses, including a rat
polyomavirus and mumps virus (MuV; Mononegavirales:
Paramyxoviridae: Mumps rubulavirus)48–50.
With the exception of the adrenal cortex, infection of the
endocrine system by EBOV has received little atten-
tion30,31,33,41. EBOV infection of the thyroid follicles and
pancreatic islets has been described in rhesus monkeys
with a prolonged clinical course, and previous guinea pig
studies have reported virus without lesions in the thy-
roid interstitium30,51. Infection of the pancreatic islets and
adrenal cortex and medulla has been reported during
acute Marburg virus infection in rhesus monkeys and
humans52,53. Virally-mediated destruction of these tis-
sues could potentially predispose to autoimmune disease. While
endocrine disorders have not been specifically
reported among EVD survivors, disease presentation can be
17. protean, and diagnosis difficult.
Investigations to see whether the infected tissues and cell types
identified in our study also are infected in
other animal models of EVD and whether filoviruses other than
EBOV mimic the observed tropism in the vari-
ous animals will be interesting. Discrepancies could possibly
explain the differences between the filovirus disease
animal models in terms of possible routes of infection,
incubation period, presence or absence of individual
clinical signs, lethality, and efficacy of candidate medical
countermeasures. That said, our study has several lim-
itations. First, we retrospectively analyzed tissues collected
during a standard EBOV confirmation-of-virulence
experiment. Thus, none of the examined tissues were target
tissues for collection. Instead, the tissues were col-
lected incidentally with other tissues expected to produce
typical lesions of EBOV infection (e.g., liver, lymphoid
organs, adrenal cortex), resulting in non-uniform sampling. A
more thorough guinea pig study therefore ought
to be performed with the sole focus to identify all EBOV target
cells in vivo. Second, none of the collected tissues
were analyzed by ISH for the presence of EBOV antigenomes,
thereby proving EBOV replication, nor were they
examined by transmission electron microscopy or for virus
titers. This latter omission may have contributed to
our limited ability to make correlations between infectious
inoculum dose and lesion development. At this point,
we therefore do not know whether the infection of the various
newly identified EBOV-susceptible cell types truly
leads to production of progeny virions, and consequently we
cannot speculate about how much our findings truly
contribute to pathogenesis. However, results of our study
indicate that the pathology of filovirus infections in
animal models is much less understood than previously
appreciated, and that multiple opportunities may exist
18. for filoviruses to establish cryptic and/persistent infections
throughout the infected body.
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Materials and Methods
Animals and virus. Fifteen male and 15 female Hartley guinea
pigs (Cavia porcellus, Crl:HA, Charles River
Laboratories, Wilmington, MA, strain code 051) were
acclimatized to the Maximum Containment (Biosafety
Level 4) Laboratory at the US National Institutes of Health
(NIH)/National Institute of Allergy and Infectious
Diseases (NIAID)/Division of Clinical Research
(DCR)/Integrated Research Facility at Fort Detrick (IRF-
Frederick). The guinea pigs were randomized to one of three
dosage groups (n = 10/group) and infected by
the intraperitoneal route with guinea pig-adapted Ebola
virus/UTMB/C.porcellus-lab/COD/1976/Yambuku-
Mayinga-GPA (BioSample ID: SAMN05755726; henceforth
EBOV), receiving the targeted doses of 10, 100, or
1,000 PFU, respectively.
All work with infectious virus was conducted in a Maximum
Containment (Biosafety Level 4) Laboratory at
the IRF-Frederick that is fully accredited by the Association for
the Assessment and Accreditation of Laboratory
Animal Care International. All animal experiments were
performed in accordance with animal study protocols
approved by a DCR Animal Care and Use Committee. Protocols
were compliant with the US Department of
Agriculture Animal Welfare Act regulations and the US Public
19. Health Service Policy on the Humane Care and
Use of Laboratory Animals and adhered to the recommendations
stated in The Guide for the Care and Use of
Laboratory Animals.
Twenty-three of the 30 guinea pigs were humanely euthanized
in accordance with defined experimental end-
points (Table 1) between d 7 and 9 post-exposure, and gross
necropsy was performed by one of two American
College of Veterinary Pathologists (ACVP) diplomate
veterinary pathologists (SY and LH). Seven guinea pigs
succumbed to disease and were not necropsied due to autolysis.
Tissues were fixed for 72 h in 10% neutral buff-
ered formalin before automated processing in a Tissue-Tek VIP-
6 vacuum infiltration processor (Sakura Finetek
USA, Torrance, CA) followed by paraffin embedding with a
Tissue-Tek model TEC (Sakura). Slides were cut on
a Leica model 2245 microtome at 4 µm, stained with
hematoxylin and eosin (H&E) and coverslipped. Slides were
examined by a single ACVP diplomate veterinary pathologist
(TKC) blinded to intervention. All images were
captured with a Leica DM3000 microscope and DFC 500 digital
camera using Leica Application Suite version
4.10.0 (Leica Microsystems, Buffalo Grove, IL).
Immunohistochemistry. EBOV immunohistochemistry (IHC)
was performed with mouse anti-EBOV
matrix protein (VP40) antibody (1:1500; 3G5, catalog 0201–
016; IBT Bioservices, Rockville, MD) or rabbit
anti-EBOV glycoprotein (GP1,2) antibody (1:14,000; catalog
0301-015; IBT Bioservices), followed by biotiny-
lated anti-mouse (catalog 115-065-166, Jackson
Immunoresearch Laboratories, West Grove, PA) or anti-rabbit
secondary antibody (catalog 111-065-144, Jackson
Immunoresearch Laboratories), and an avidin-biotin perox-
idase tertiary antibody (catalog PK-6100; Vector Laboratories,
20. Burlingame, CA). Staining was visualized with
3,3′-diaminobenzidine (DAB) chromogen (catalog BDB2004L;
Biocare Medical, Concord, CA) and counter-
stained with hematoxylin.
In Situ Hybridization. EBOV RNA in situ hybridization (ISH) in
formalin-fixed, paraffin-embedded
(FFPE) tissues was performed using the RNAscope 2.5 high
definition (HD) RED kit (Advanced Cell
Diagnostics, Newark, CA) according to the manufacturer’s
instructions26. Briefly, 20 ZZ probe pairs targeting
the genomic EBOV nucleoprotein (NP) gene were designed and
synthesized by Advanced Cell Diagnostics (cat-
alogue 448581). After deparaffinization with xylene, a series of
ethanol washes and peroxidase blocking, sec-
tions were heated in Antigen Retrieval Buffer (Advanced Cell
Diagnostics) and then digested by proteinase K
(Advanced Cell Diagnostics). Sections were exposed to ISH
target probe and incubated at 40 °C in a hybridi-
zation oven (HybEZ™, Advanced Cell Diagnostics) for 2 h.
After rinsing, the ISH signal was amplified using
company-provided Pre-amplifier and Amplifier conjugated to
alkaline phosphatase (AP) and incubated with
a red substrate-chromogen solution for 10 min at room
temperature. Sections were then counterstained with
hematoxylin, air-dried, and coverslipped.
Data availability. Digitally scanned slides will be made
available upon request.
Score Criteria*
0 No evidence of pain or distress. Normal activity.
1
Mild or anticipated pain or distress. Examples of clinical signs:
hiding or decreased activity, soft
21. tissue swelling or discomfort without other signs, abnormal or
hunched posture, or unthrifty,
ungroomed or scruffy appearance, but active.
2
Moderate pain or distress. Examples of clinical signs: rapid or
shallow respiratory rate, head tilt,
paresis (limb weakness, difficulty in ambulation), abnormal
nesting behavior, abnormal or hunched
posture, or unthrifty, ungroomed or scruffy appearance with
reduced activity, but responsive when
stimulated.
3
Severe pain or distress. Examples of clinical signs: pale skin or
mucous membranes, unable or
unwilling to move or comatose (moribund) when stimulated,
agonal breathing, paralysis, head
tilt with circling or rolling, persistent scratching, abnormal or
hunched posture or unthrifty, or
ungroomed or scruffy appearance.
Table 1. Experimental endpoint criteria. *Scoring is based on
the presence of one or more clinical signs within
the example list. Guinea pigs were considered moribund and
were euthanized after reaching a score of 3.
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Acknowledgements
The content of this publication does not necessarily reflect the
views or policies of the US Department of the
Army, the US Department of Defense, or the US Department of
Health and Human Services or of the institutions
and companies affiliated with the authors. This work was
funded in part through Battelle Memorial Institute’s
prime contract with the US National Institute of Allergy and
Infectious Diseases (NIAID) under Contract
No. HHSN272200700016I. J.C.J. and J.H.K. performed this
work as employees of Battelle Memorial Institute.
Subcontractors to Battelle Memorial Institute who performed
this work are: T.K.C., L.H., O.R., A.B., R.H., and
S.Y., employees of Charles River Laboratories. The authors
thank Dr. Thomas W. Geisbert (University of Texas
Medical Branch, Galveston, TX, USA) for the kind gift of Ebola
virus/UTMB/C.porcellus-lab/COD/1976/
Yambuku-Mayinga-GPA. The authors would like to thank IRF-
Frederick staff Jiro Wada for figure preparation
and Dr. Donna Perry and Laura Bollinger for critically
reviewing and editing the manuscript.
Author Contributions
Conceptualization: T.K.C., J.C.J., P.B.J., J.H.K., X.Z.;
Methodology: T.K.C., L.H., J.C.J., O.R., S.Y., A.B., R.H., X.Z.;
Formal analysis: T.K.C., X.K.; Writing - original draft: T.K.C.,
J.H.K.; Writing - review & editing: all authors;
Supervision: M.G.S., S.B., P.B.J.; Project administration:
J.C.J.; Funding acquisition: P.B.J.
Additional Information
Supplementary information accompanies this paper at
33. http://dx.doi.org/10.1177/0192623317723541
http://dx.doi.org/10.1086/520589
http://dx.doi.org/10.1086/520608
http://dx.doi.org/10.1038/s41598-018-19638-x
http://creativecommons.org/licenses/by/4.0/Histology,
immunohistochemistry, and in situ hybridization reveal
overlooked Ebola virus target tissues in the Ebola virus
...Results
Liver. Heart. Nerves and ganglia. Genital tract. Salivary glands.
Thyroid. Adrenal Glands. Pancreas. Discussion
Materials and Methods
Animals and virus. Immunohistochemistry. In Situ
Hybridization. Data availability. Acknowledgements
Figure 1 EBOV infection of the liver and heart.Figure 2 EBOV
infection of paradrenal ganglia and peripheral medium
myelinated nerves.Figure 3 EBOV infection of the uterus.Figure
4 EBOV infection of the vagina, penis, and prepuce.Figure 5
EBOV infection of the salivary glands and thyroid.Figure 6
EBOV infection of adrenal medulla and pancreas.Table 1
Experimental endpoint criteria.