Ch. 2, 3, 4 Genomics, Stems Cells, PRP

Question 1

what does mRNA code for

Answer 1

messenger RNA which codes for protein sequences

Question 2

what does rRNA code for

Answer 2

ribosomal RNA codes for mitochondrial ribosomal subunits involved in translation

Question 3

what does tRNA code for?

Answer 3

transfer RNA which codes for amino acid binding units that bind to mRNA molecules

Question 4

what does small nuclear RNA code for?

Answer 4

the units of the spliceosomes, the complex of RNA and protein that removes the introns from transcribed RNA

Question 5

What does small nucleolar RNA code for

Answer 5

molecules involved in RNA modification

Question 6

What mediates the transcription process? and where does it primarily occur?

Answer 6

DNA directed RNA polymerase

in the nuclei of eukaryotic cells and a little in the mitochondria

Question 7

What elements of DNA are highly conserved between mammalian species

Answer 7

non coding segments - they are usually coding for regulation of development

Question 8

what is reverse transcriptase

Answer 8

a protein that can generate a DNA sequence from an RNA template

Question 9

what are retrotransposons

Answer 9

sequences of DNA that can move around the genome of a cell
They are transcribed to mRNA and then back to DNA using reverse transcriptase
the DNA then can be integrated back into the genome and that may result in mutations
ex. long terminal repeats (similar to retroviruses), short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs)

Question 10

What is an example of a transposon causing disease in a dog?

Answer 10

• Lamellar ichthyosis in Jack Russell terriers occurs following insertion of a LINE sequence into intron 9 of the transglutaminase 1 gene (TGM1)
• heredity myopathy in Labrador retrievers is caused by a tRNA derived SINE positioned in exon 2 of the protein tyrosine phosphatase like member A (PTPLA)

Question 11

What four transcription factors will reprogram human somatic cells into functional pluripotent stem cells

Answer 11

OCT4
SOX2
NANOG
LIN28

Question 12

what is the imprinting of genes

Answer 12

the expression of only a single allele of a gene of the two copies inherited by the parents, rather than both copies

Question 13

what does a western blot measure

Answer 13

detects specific proteins

detect small quantities of proteins in limited samples

Question 14

what does an ELISA measure

Answer 14

detects antigens or antibodies
can detect almost any antigen for which the corresponding antibody is available or vice versa
rapid and suited for high throughput screening

Question 15

what is the difference between a prognostic factor and a predictive factor

Answer 15

a prognostic factor is a measure of the natural history of the disease. it is most appropriately assessed in a control group from a randomized clinical trial
a predictive factor suggests that the differential benefit of therapy is dependent on the status of the predictive marker

Question 16

What causes the MDR1 mutation toxicity

Answer 16

in collies, a 4 base pair deletion in MDR1 gene leads to a stop codon that prematurely terminates MDR1 protein synthesis
MDR1 protein is a large transmembrane P glycoprotein that helps in drug transport at the blood brain barrier
with abnormal MDR1, ivermectin will accumulate in the brain

Question 17

what are the three broad classes of stem cells

Answer 17

embryonic stem cells
adult stem cells
fetal/perinatal stem cells

Question 18

what does totipotent mean

Answer 18

potential to differentiate into cells of each germ layer (ectoderm, endoderm, mesoderm) as well as trophodectoderm (extraembyronic tissue)

Question 19

what does pluripotent mean

Answer 19

potential to differentiate into the three germ layers but not the extraembryonic tissues

Question 20

what does multipotent eman

Answer 20

differentiate into cells within a single germ cell layer lineage

Question 21

what are the criteria for calling something a mesenchymal stem cell by the Dominici et al 2006 paper

Answer 21

1. adhere to tissue culture plastic and exhibit a spindle shape
2. form colonies of cells from single parent cells when cultured in low density
3. express a specific set of cell marker proteins that exclude the cells from hematopoietic lineages
4. possess the ability to differentiate into osteoblast, adipocytes, and chondrocytes

Question 22

what are some antiinflammatory agents produced by mesenchymal stem cells

Answer 22

TNF alpha-stimulated gene/protein 6 (TSG-6)
interleukin 1 receptor antagonist
Prostaglandin E2

Question 23

what are the steps of the mesengenic process of stem cell differentiation?

Answer 23

1. proliferation
2. commitment
3. lineage progression
4. differentiation
5. maturation

Question 24

what would be the most readily accessible stem cells on the clinic floor?`

Answer 24

bone marrow concentrate aka stromal vascular fraction using systems from MediVet or InGeneron or Vetbiologics
these have a minor population of stem/progenitor cells along with other nucleated cells
outcomes might not be predictable because who knows what percentage of stem cells you are putting in

Question 25

How are veterinary mesenchymal stem cells administered locally to a site of injruy

Answer 25

either in a soluble solution, injected in conjunction with a platelet rich plasma or fibrin gel, attached to a scaffold/matrix

Question 26

what has direct IV injection of mesenchymal stem cells been associated with

Answer 26

myocardial infarction or ischemia
pulmonary edema
hemorrhage

Question 27

what protein is necessary to maximize dog osteogenic differentiation from stem cells?

Answer 27

BMP 2 or bone morphogenic protein 2

Question 28

What should the concentration of platelets be to be considered PRP

Answer 28

at least 3 to 5 times higher than the concentration in peripheral blood

Question 29

What is IRAP

Answer 29

interleukin 1 receptor antagonist or interleukin 1 receptor protein will competitively inhibit IL 1 beta activity
collected with a conditioner and must be incubated for several hours
like PRP, concentrations are variable in your end product