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More presentations at http://www.swinecast.com/2016-missouri-pork-expo
Dr. Randy Prather - Porcine Reproductive and Respiratory Syndrome Virus Resis...John Blue
Porcine Reproductive and Respiratory Syndrome Virus Resistant Pigs - Dr. Randy Prather, University of Missouri, from the 2015 North American PRRS Symposium, December 4 - 5, 2015, Chicago, IL, USA.
More presentations at http://www.swinecast.com/2015-north-american-prrs-symposium
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Resilience and PRRS in a natural disease challenge model - Dr. Graham Plastow, University of Alberta, from the 2017 North American PRRS/National Swine Improvement Federation Joint Meeting, December 1‐3, 2017, Chicago, Illinois, USA.
More presentations at http://www.swinecast.com/2017-north-american-prrs-nsif-joint-meeting
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I cover
1. What biological data looks like today
2. How the revolution in genomics sequencing technology is IN a hospital near you
3. How this is affecting patient treatment today
4. What are some of the major challenges in using this data in the clinic?
and ...
5. (1 slide about ) How my research fits into the paradigm of understanding human genetic variation.
Supporting Genomics in the Practice of Medicine by Heidi RehmKnome_Inc
View the webinar at http://www.knome.com/webinar-supporting-genomics-practice-medicine. In this presentation, Dr. Heidi Rehm, Chief Laboratory Director of the Laboratory for Molecular Medicine at Partners Healthcare and one of the Principal Investigators on ClinGen, elucidates the challenges of genomics in medicine and outlined the path to integrating large scale sequencing into clinical practice.
Dr. Randy Prather - Porcine Reproductive and Respiratory Syndrome Virus Resis...John Blue
Porcine Reproductive and Respiratory Syndrome Virus Resistant Pigs - Dr. Randy Prather, University of Missouri, from the 2015 North American PRRS Symposium, December 4 - 5, 2015, Chicago, IL, USA.
More presentations at http://www.swinecast.com/2015-north-american-prrs-symposium
Dr. Graham Plastow - Resilience and PRRS in a natural disease challenge modelJohn Blue
Resilience and PRRS in a natural disease challenge model - Dr. Graham Plastow, University of Alberta, from the 2017 North American PRRS/National Swine Improvement Federation Joint Meeting, December 1‐3, 2017, Chicago, Illinois, USA.
More presentations at http://www.swinecast.com/2017-north-american-prrs-nsif-joint-meeting
Big data biology for pythonistas: getting in on the genomics revolutionDarya Vanichkina
Slides for the talk I gave at PyCon Australia trying to simplify biology and genomics into something easily accessible for software developers and CompSci graduates.
I cover
1. What biological data looks like today
2. How the revolution in genomics sequencing technology is IN a hospital near you
3. How this is affecting patient treatment today
4. What are some of the major challenges in using this data in the clinic?
and ...
5. (1 slide about ) How my research fits into the paradigm of understanding human genetic variation.
Supporting Genomics in the Practice of Medicine by Heidi RehmKnome_Inc
View the webinar at http://www.knome.com/webinar-supporting-genomics-practice-medicine. In this presentation, Dr. Heidi Rehm, Chief Laboratory Director of the Laboratory for Molecular Medicine at Partners Healthcare and one of the Principal Investigators on ClinGen, elucidates the challenges of genomics in medicine and outlined the path to integrating large scale sequencing into clinical practice.
Alpha-1 Antitrypsin (α-1 AT) deficiency is a common genetic disorder that affects 1 in 2,000 individuals in the USA. Additionally, over 20 million people have been identified as carriers for this genetic disorder. In severe cases, α-1 AT deficiency can cause substantial lung and liver damage, which if left untreated could result in death and there are no current available treatments. Alpha-1 protein is produced in the liver, travels in the bloodstream and utilized in the lungs to protect healthy lung tissue from harmful destruction by elastase. A common single amino acid substitution, located at E342K (ATZ) was identified in α-1 AT deficient humans. When this specific mutation occurs two phenotypes can result: 1) ATZ can polymerize in the liver causing cellular toxicity 2) inhibits alpha-1 antitrypsin from inhibiting elastase which can result in lung disease. Currently; little is known about the cellular mechanisms that clear the accumulated proteins in the liver. Therefore, an investigative study utilizing C. elegans model of ATZ was performed in order to help determine the cellular mechanisms that dispose of accumulated proteins. Specifically RNA interference was utilized to knockdown expression of specific genes. This investigation examined genes involved in the heat-shock pathway (HSP), unfolded protein response (UPR), and insulin signaling pathway (IS). Phenotypic analysis including: embryonic lethality, protein aggregation expression, and longevity, was completed after knockdown of genes to determine effect on ATZ accumulation. Currently with our preliminary data suggests that the heat-shack pathway may play a role in ATZ accumulation. Determining the mechanism of protein accumulation in the investigation of C. elegans may lead to possible drug targets and therefore the development of a treatment which may alleviate those diagnosed with this disorder.
CONCEPT
HISTORY OF XENOTRANSPLANTATION
IMPORTANCE OF XENOTRANSPLANTATION
CHOOSING OF DONOR SPECIES
XENOTRANSPLANTATION REJECTION
GUIDELINES ON XENO-TRANSPLANTATION BY ICMR
RECENT RESEARCH ON XENOTRANSPLANTATION
Dr. Ben Hause - Pathogen Discovery Using Metagenomic SequencingJohn Blue
Pathogen Discovery Using Metagenomic Sequencing - Dr. Ben Hause, College of Veterinary Medicine, Kansas State University, from the 2016 Allen D. Leman Swine Conference, September 17-20, 2016, St. Paul, Minnesota, USA.
More presentations at http://www.swinecast.com/2016-leman-swine-conference-material
Dr. Ben Hause - Metagenomic Sequencing for Virus Discovery and CharacterizationJohn Blue
Metagenomic Sequencing for Virus Discovery and Characterization - Dr. Ben Hause, Kansas State University, from the 2015 North American PRRS Symposium, December 4 - 5, 2015, Chicago, IL, USA.
More presentations at http://www.swinecast.com/2015-north-american-prrs-symposium
Presented by Etienne de Villiers at the African Swine Fever Diagnostics, Surveillance, Epidemiology and Control Workshop, Nairobi, Kenya, 20-21 July 2011
Novel hexavalent GITR agonists stimulate T cells and enhance memory formationThomas Hoeger
Apogenix's novel hexavalent GITR agonists as candidates for immunotherapeutic treatment of cancer – look at the presentation of comprehensive data by Meinolf Thiemann, PhD, at the AACR Annual Meeting in Washington, D.C., on April 4, 2017.
Alpha-1 Antitrypsin (α-1 AT) deficiency is a common genetic disorder that affects 1 in 2,000 individuals in the USA. Additionally, over 20 million people have been identified as carriers for this genetic disorder. In severe cases, α-1 AT deficiency can cause substantial lung and liver damage, which if left untreated could result in death and there are no current available treatments. Alpha-1 protein is produced in the liver, travels in the bloodstream and utilized in the lungs to protect healthy lung tissue from harmful destruction by elastase. A common single amino acid substitution, located at E342K (ATZ) was identified in α-1 AT deficient humans. When this specific mutation occurs two phenotypes can result: 1) ATZ can polymerize in the liver causing cellular toxicity 2) inhibits alpha-1 antitrypsin from inhibiting elastase which can result in lung disease. Currently; little is known about the cellular mechanisms that clear the accumulated proteins in the liver. Therefore, an investigative study utilizing C. elegans model of ATZ was performed in order to help determine the cellular mechanisms that dispose of accumulated proteins. Specifically RNA interference was utilized to knockdown expression of specific genes. This investigation examined genes involved in the heat-shock pathway (HSP), unfolded protein response (UPR), and insulin signaling pathway (IS). Phenotypic analysis including: embryonic lethality, protein aggregation expression, and longevity, was completed after knockdown of genes to determine effect on ATZ accumulation. Currently with our preliminary data suggests that the heat-shack pathway may play a role in ATZ accumulation. Determining the mechanism of protein accumulation in the investigation of C. elegans may lead to possible drug targets and therefore the development of a treatment which may alleviate those diagnosed with this disorder.
CONCEPT
HISTORY OF XENOTRANSPLANTATION
IMPORTANCE OF XENOTRANSPLANTATION
CHOOSING OF DONOR SPECIES
XENOTRANSPLANTATION REJECTION
GUIDELINES ON XENO-TRANSPLANTATION BY ICMR
RECENT RESEARCH ON XENOTRANSPLANTATION
Dr. Ben Hause - Pathogen Discovery Using Metagenomic SequencingJohn Blue
Pathogen Discovery Using Metagenomic Sequencing - Dr. Ben Hause, College of Veterinary Medicine, Kansas State University, from the 2016 Allen D. Leman Swine Conference, September 17-20, 2016, St. Paul, Minnesota, USA.
More presentations at http://www.swinecast.com/2016-leman-swine-conference-material
Dr. Ben Hause - Metagenomic Sequencing for Virus Discovery and CharacterizationJohn Blue
Metagenomic Sequencing for Virus Discovery and Characterization - Dr. Ben Hause, Kansas State University, from the 2015 North American PRRS Symposium, December 4 - 5, 2015, Chicago, IL, USA.
More presentations at http://www.swinecast.com/2015-north-american-prrs-symposium
Presented by Etienne de Villiers at the African Swine Fever Diagnostics, Surveillance, Epidemiology and Control Workshop, Nairobi, Kenya, 20-21 July 2011
Novel hexavalent GITR agonists stimulate T cells and enhance memory formationThomas Hoeger
Apogenix's novel hexavalent GITR agonists as candidates for immunotherapeutic treatment of cancer – look at the presentation of comprehensive data by Meinolf Thiemann, PhD, at the AACR Annual Meeting in Washington, D.C., on April 4, 2017.
A holistic approach to control of pig diseases 29.05.15Joseph Kungu
Pigs are affected by a variety of conditions which directly kill or reduce their productivity.
A number of these infections are resident in Uganda. However, not much studies done.
Dr. Jack Dekkers - Genetics of Host Resistance to PRRS and PCV2John Blue
Genetics of Host Resistance to PRRS and PCV2 - Dr. Jack Dekkers, Iowa State University, from the 2015 North American PRRS Symposium, December 4 - 5, 2015, Chicago, IL, USA.
More presentations at http://www.swinecast.com/2015-north-american-prrs-symposium
ABSTRACT- The biochemical and histopathological changes in rats experimentally infected with T. evansi isolated from camels in El-Gadarif State, Sudan, were studied. A number of 18 adult male outbred albino rats, weighing between 133-137g were used in the study. The rats were divided into 3 groups of 6 animals each (A,B and E). Group A and B were intraperitoneally infected with T. evansi (Showak stabilate) with 1×104 trypanosoma for the inoculum. Group B was given quinapyramine sulphate (20 mg/kg bwt) after parasitaemia was evident. Group E was left healthy uninfected controls for the stabilate. There was significant reduction in serum glucose and phosphorus; compared to significant increase in Glutamate Oxaloacetate Transaminase (GOT), Glutamate Pyruvate Transaminase (GPT) and total protein in groups (A and B). Microscopically, the brain tissues of the infected rats revealed acute congestion of the meningeal capillaries, perivascular oedema, neuronecrosis (vaculation), gliosis and trypomastigotes in dilated capillaries. The lung revealed oedema, congestion, multifocal alveolar emphysema, hyperplasia of the peri-bronchiolar lymphoid tissues and haemorrhages. The spleen showed extensive haemorrhages, haemosiderosis and aggregation of histiocytes resulting in multinuclear giant cells formation. The kidneys showed acute congestion of the glomerular tufts. All tissues obtained showed exactly the same histopathological changes. No significant histopathological alterations were observed in the liver and heart. The most consistent histopathological changes were seen in the brain, lungs, spleen and kidneys. These changes were consistent with trypanosome infection and were confirmed by the presence of trypanosomes in most of the tissue sections examined.
Key-words- histopathological, biochemical, changes, T. evansi, dromedary camels, Sudan
Dr. Dyneah Classen - Management and challenges of dealing with swine influenzaJohn Blue
Management and challenges of dealing with swine influenza - Dr. Dyneah Classen, Carthage Veterinary Service, Ltd., from the 2013 Allen D. Leman Swine Conference, September 14-17, 2013, St. Paul, Minnesota, USA.
More presentations at http://www.swinecast.com/2013-leman-swine-conference-material
Investigations for iufd & sb, how to select?Wafaa Benjamin
Foetal loss is a distressing situation for the lady ,family and medical staff as well.
Investigating the cause of death has many benefits .
Meticulous history taking and clinical assessment is of at most importance.
There are routine standard tests & others arte selective directed by clinical scenarios.
Researches & recording are required to estimate main causes of foetal death at local level, so, investigations could be directed.
In presence of lack of resources, selection of investigations should be prioritized by most relevant and most informative ones.
Post-mortem examination should be re-included at least external examination & placental histopathology.
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2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
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6. • The PRRS virus (PRRSv) was first detected in
the U.S. in 1987 (Keffaber et al ‘89) and in Europe in
1990 (Wensvoort et al ’91)
• PRRSv replicates in macrophages
– induce prolonged viremia and can cause
persistent infections that last for months
• It also predisposes infected pigs to other
bacterial and viral pathogens
7. • Vaccinations to date have been ineffective
– Non- neutralizing antibodies actually enhance viral
replication in alveolar macrophages
– Antibody dependent enhancement (ADE)
• Sows/Gilts
– severe reproductive failure and a high rate of late abortion
– Early farrowing
– Decreased litter size, Increased number of mummies
• Boars
– Low libido
– Fever, low sperm count
• Young and growing pigs
– Pneumonia, Diarrhea, Mortality of 12-15%
http://www.thepigsite.com/diseaseinfo/97/porcine-reproductive-respiratory-syndrome-prrs
8. • Costs $660,000,000 annually in North
America (Holtkamp et al ‘13)
• Costs €1,500,000,000 annually in
Europe (European PRRSpecive ‘15)
• Translates to ~$6,000,000 each day!
• Doesn’t include Asia.
10. – Initial binding with heparan sulfate
– Binding/Internalization by Sialoadhesin
– Internalization/uncoating of the virus by CD163
Van Breedam et al.,
2010
16. • Piglets were shipped to Kansas State
University
• After 1 week acclimation
– Low passage PRSSV isolate (KS-06: North
American isolate collected in 2006)
– 105 TCID50 of virus diluted in 3 mL
• ½ intramuscularly
• ½ intranasally
– Blood sampled on days 0, 4, 7, 14, 21, 28 &
35.
Prather et al ‘13 J Virology
18. • SIGLEC1-/- is not embryonic lethal.
• SIGLEC1-/- did not affect susceptibility
to PRRSv.
Prather et al ‘13 J Virology
19. • Bacterical immune system that
degrades foreign genetic elements
(plasmids, phages).
• Can edit the genome at a very high
efficiency.
• Uses two main components: a guide
sequence and a Cas9 protein.
20.
21. • Non-Homologous End Joining
• Can result in changing a handful of
base pairs
• Thus can remove exons/domains or
knockout a gene.
• Swine genome is 2,700,000,000+ base
pairs.
23. • 3 piglets predicted to be null (CD163-/- )
• 8 WT piglets
– One WT piglet humanely euthanized on day 1, due to poor body
condition and not included in the study
• K-State veterinarians and staff blinded to
genotypes
Piglet ID Predicted
Translation
Maternal Allele Paternal Allele
#43 CD163-/- 7 bp addition in exon 7 2 bp deletion in exon 7 + 377
bp intron deletion in the
preceding intron
#55 CD163-/- 7 bp addition in exon 7 2 bp deletion in exon 7 + 377
bp intron deletion in the
preceding intron
#40 CD163-/- 7 bp addition in exon 7 11 bp deletion in exon 7
Whitworth et al, ‘15
Nature Biotechnology
24. • Identity blinded to the crew at KSU
• Housed in the K-State BL-2 LARC facility
• Allowed to acclimate for 3 days after arrival at facility
• Challenge IM and IN with 105 TCID50 of NVSL 97-7985.
(Standard lab isolate from 1997, relatively “hot” in terms of
replication and pathogenesis)
• Pigs were maintained in the same pen; therefore, all pigs
constantly exposed to virus
• Monitored daily for clinical signs
• Blood collected on days 0,4,7,11,14,21,28,35 and weights
collected at least weekly
• Study terminated 35 days after infection
• Lung lavage for PAMs
• Lungs and tissues removed for histopathology
Whitworth et al, ‘15
Nature Biotechnology
26. • Fever was considered positive if it was ≥ 104°F
• Respiratory scores ranged from:
– 0: Normal, to
– 1: mild dyspnea and/or tachypnea when stressed
(when handled),
– 2: mild dyspnea and/or tachypnea when at rest,
– 3: moderate dyspnea and/or tachypnea when stressed
(when handled),
– 4: moderate dyspnea and/or tachypnea when at rest,
– 5: severe dyspnea and/or tachypnea when stressed
(when handled),
– 6: severe dyspnea and/or tachypnea when at rest. Is
there evidence of diarrhea (grade) or vomiting?
Whitworth et al, ‘15
Nature Biotechnology
31. • CD163+/+ pigs had interstitial edema with the infiltration of
mononuclear cells and the mononuclear infiltrate consisted
mainly of lymphocytes and plasma cells and lesser numbers of
macrophages. In contrast there was no evidence for pulmonary
changes in the CD163-/- pigs.
Whitworth et al, ‘15
Nature Biotechnology
32. Pig Genotype Description Score*
41 Wild Type 100% congestion. Multifocal areas of edema. Infiltration of moderate numbers
of lymphocytes and macrophages.
3
42 Wild Type 100% congestion. Multifocal areas of edema. Infiltration of moderate numbers
of lymphocytes and macrophages.
3
47 Wild Type 75% multifocal infiltration with of mononuclear cells and mild edema. 2
50 Wild Type 75% moderate infiltration of mononuclear cells within alveolar spaces and
around small blood vessels. Perivascular edema.
3
51 Wild Type 25% atelectasis with moderate infiltration of mononuclear cells. 1
52 Wild Type 10% of alveolar spaces collapsed with infiltration of small numbers of
mononuclear cells.
1
56 Wild Type 100% diffuse moderate interstitial infiltration of mononuclear cells.
Interalveolar septae moderately thickened by hemorrhage and edema.
4
40 CD163-/- No changes 0
43 CD163-/- No changes 0
55 CD163-/- No changes 0
*Lung lesion scores calculated according to Halbur et al. 8 Vet Pathol. 1995 Nov;32(6):648-60
Whitworth et al, ‘15
Nature Biotechnology
33. • SIGLEC1-/- pigs are not resistant to
PRRSv.
• CD163 -/- pigs are resistant to PRRSv.
• Working on challenging other
genotypes.
• Working on challenging with other
isolates (multiple Type 1 and Type 2).
35. • Technology Permits Answering Basic
and Applied Questions about Human
Medicine and Domestic Animal
Biology
• Any Genetic Modification that you can
imagine, we can probably build.
– Miniature Pig
– Domestic Pig
• CD163-/- Entry Into Production
– Growth and Performance
36. Zygote
Injections
Kiho Lee
Lee Spate
CRISPR and Targeting Vector
Design
Kevin Wells
Kristin Whitworth
Transfection and
Genotyping
Mykel Anderson
Mariah Thomas
Joshua Benne
Nuclear Transfer and Embryo
Transfer
Joshua Benne
Stephanie Murphy
Jennifer Teson
Jiude Mao
Clifton Murphy
Surrogate and Piglet Care
Melissa Samuel
Jason Dowell
Tricia Meyer
Entire Prather Lab
Funding Sources
Genus plc
The Christopher Columbus Fellowship
Foundation
Food for the 21st Century
USDA ARS
Genus plc
Alan Mileham
Dave McLaren
Jon Lightner
SIGLEC1-/-
Project
Jon Green
Tina Egen
Kansas State University
Bob Rowland
Benjamin Trible
Maureen Kerrigan
Catherine Ewen
Ada Cino-Ozuna
Bhupinder Bawa