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Toxicology Final > Molecular/Toxicology Genotoxicity > Flashcards

Molecular/Toxicology Genotoxicity Flashcards

1
Q

What are primary tools in toxicology?

A

Chemistry, biochemistry, and molecular biology.

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2
Q

What tools does chemistry have to offer?

A

Analytical methods for toxicants and their metabolites.

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3
Q

What tools does biochemistry have to offer?

A

methods for investigating metabolism and

modes of toxic action.

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4
Q

What tools does molecular biology have to offer?

A

methods for investigating
the roles of genes and gene expression in
toxicology.

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5
Q

Who and when was the DNA structure discovered?

A

Watson and Crick in 1953

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6
Q

What occurred in the 1960s?

A

The definition of the genetic code.

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7
Q

Molecular toxicology is what?

A

restricted to the events involving DNA, RNA, Proteomics, and Metabolomics.

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8
Q

What is the goal of molecular toxicology?

A

define underlying molecular mechanisms of toxicity.

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9
Q

What do molecular techniques help you understand?

A

the mechanisms and responses of an organism to a toxic insult.

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10
Q

What areas does genetic toxicology work with?

A

Cytotoxicity, mutations, cytogenetic abberations, DNA damage and repair, and carcinogenesis.

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11
Q

What are the areas that molecular toxicology encompass?

A

Genetic toxicology, Epigenetics, bioinformatics, Toxicogenomics, Proteomics, metabolomics, gene expression, and gene function.

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12
Q

What are the 3 main areas of molecular toxicology needed to know for class?

A

Genetic tox (mutations and cytogenetic aberrations), toxicogenomics, and gene expression.

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13
Q

What is genetic toxicology?

A

Study and characterization of physical and chemical agents that affect the hereditary material of living organisms.

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14
Q

What are hereditary materials?

A

DNA and RNA

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15
Q

What is the primary goal of a genetic toxicologist?

A

Detect and understand the properties of agents that induce deleterious effects in genetic elements at doses below those that induce physiological toxicity.

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16
Q

How did the field of genetic toxicology originate?

A

With the work of HJ muller and C Auerbach.

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17
Q

Who was HJ Muller?

A

Radiation mutagenesis

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18
Q

Who was C. Auerbach?

A

Chemical mutagenesis.

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19
Q

What occurred to Muller in 1927?

A

He discovered ionizing radiation, specifically X-rays, induced mutations in drosophila

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20
Q

Who publicized the adverse implications of inappropriate use of medical x-rays?

A

HJ Muller in 1930

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21
Q

What were X-rays being used for in 1930s?

A

To xrays your feet inside the shoe to be sure it fit well.

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22
Q

In 1946 What did Charlotte Auerbach discover?

A

chemicals could induce mutations in Drosophila.

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23
Q

What type of chemicals did C. Aurbach investigations used?

A

Mustard Gas use in WW2

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24
Q

C. Auerbach was one of the first scientist to consider what?

A

Chemical mutagens could induce genetic damage in humans.

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25
Q

What did the two major advances in the 1970s provide clearer understanding to?

A

mechanisms of mutagenesis and the potential role of mutagens in carcinogenesis.

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26
Q

What did James and Elizabeth Miller demonstrate?

A

Chemical carcinogens could react to form stable covalent derivatives with DNA, RNA, Proteins.

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27
Q

Formation of derivatives required metabolisms of what?

Who discovered this?

A

The chemical to primary and subsequent metabolites.

James and Elizabeth Muller.

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28
Q

What did James and Elizabeth Muller establish?

A

Requirement of metabolism for some carcinogens.

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29
Q

What did Bruce Ames and his colleagues design?

A

An assay with Salmonella Tryphimurium to detect chemically induced reverse mutations.

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30
Q

The Ames Assay was to use to detect chemically induced reverse mutations where?

A

At the histidine locus.

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31
Q

What role does the Ames Assay play?

A

An important role in hazard identification.

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32
Q

Ames assay includes several bacteria strains that contain what?

A

A specific mutation in the histidine locus.

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33
Q

Tester strains are unable to grow in the absence of what?

A

Histidine.

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34
Q

Mutations must be reverted to normal functions why?

A

to be detected in assay (histidine independence)

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35
Q

What does TA1535 detect?

A

base pair substitutions

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36
Q

What does TA1538 detect?

A

-1 base pair frameshifts

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37
Q

What does TA1537 detect?

A

+1 base pair framshifts.

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38
Q

What is the experimental procedure for the Ames Assay?

A

Bacteria are treated with a test chemical.

Plated onto a histidine-deficient growth medium.

Colonies that develop are scored as mutants.

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39
Q

How can Ames assay also be conducted using S9?

A

(exogenous metabolizing
system) to detect mutagens that require metabolic
activation to DNA-reactive intermediate.

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40
Q

What type of approaches do we use now?

A

Omic approaches including

microarrays and whole genome sequencing.

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41
Q

What type of advances were made in mid 70s and mid 80s?

A

~200 short-term genotoxicity and
mutagenicity assays developed

Mutation induction, DNA damage,
DNA repair, cytotoxicity

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42
Q

What are three types of genetic alterations induced by a toxicant?

A

Mutagenesis, clastogenesis, Aneuploidy.

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43
Q

What is mutagenesis?

A

Loss, addition, or alteration of a small number of base pairs.

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44
Q

What is Clastogenesis?

A

Loss, addition, or rearrangement of parts of chromosomes.

45
Q

What is Aneuploidy?

A

Acquisition or loss of a complete chromosome.

46
Q

Base pair substitutions and modifications is an example of what?

A

Gene mutations: mutagenesis

47
Q

Frameshift mutations (deletions/insertions) is an example of what?

A

Mutagenesis

48
Q

Changes in the number of sets of chromosomes is an example of what?

A

Clastogenesis and aneuploidy

49
Q

Changes in the number of chromosomes is what?

A

Clastogenesis and aneuploidy

50
Q

Changes in the structure of the chromosomes (deletion, duplication, and translocation) is an example of what?

A

Clastogenesis and aneuploidy

51
Q

Substituting one base for another is what?

A

Gene mutations: remember A-T and G-C

52
Q

What can cause erroneous base pairing?

A

Incorporation of a mutagen into the DNA molecule.

53
Q

Which is worse: BP sub or Frameshift?

A

Framshift

54
Q

What is a frameshift mutation?

A

Base pair addition or deletion.

55
Q

What agents may intercalate into the DNA?

A

Ethidium bromide and acridine orange.

56
Q

Agents that intercalate into the DNA may result in what?

A

Frameshift mutation.

57
Q

In frameshift mutation what occurs to the order of the genetic code?

A

It is altered and reading from the base code is shifted.

58
Q

What is seriously affected by a frameshift mutation?

A

Transcription of information.

59
Q

What is polyploidy chromosomal damage?

A

Changes in the number of sets of chromosomes.

60
Q

What is aneuploidy chromosomal damage?

A

changes in the number of chromosomes.

61
Q

What are types of changes within the chromosome structure?

A

deletion, duplication, and translocation.

62
Q

What are genetic toxicology assays developed to screen?

A

environmental products for carcinogens.

63
Q

Genetic toxicology assays are developed to identify what?

A

putative carcinogens in body fluids.

64
Q

Genetic toxicology assays are developed to define what?

A

mechanisms of activation of chemical carcinogens and describing their reactions with DNA.

65
Q

Genetic toxicology assays are developed to understand what?

A

the process of anti-mutagenicity and anticarcinogenicity.

66
Q

Genetic toxicology assays are developed to undergo what type of research?

A

basic and molecular mutation.

67
Q

Genetic toxicology assays are developed to predict what?

A

carcinogenicity in regulatory standards for testing environmental chemicals.

68
Q

What are the ideal characteristics of a genotoxicity assay?

A

sensitivity and repoducibility and to be able to detect the entire mutagenic spectrum.

69
Q

What does it mean when a genotoxicity assay has ideal sensitivity and reproducibility?

A

ability to detect a small mutational effect above the spontaneous mutation rate.

done in any given laboratory.

70
Q

What does it mean when a genotoxicity assay has ideal capacity to detect the entire mutagenic spectrum?

A

Ability to detect changes in chromosomes number and induction of chromosomes aberrations and rearrangements. –> ability to detect single gene mutations and DNA damage.

71
Q

What are two assays used to detect gene mutations?

A

Ames assay and gene mutation assays in cell culture lines.

72
Q

How does the ames assay detect mutations?

A

Bacterial assay- Various strains to detect specific mutations

Mainly hazard identification

73
Q

How does the Gene mutation assay in cell culture lines detect mutations?

A

Target certain loci for specific mutations

74
Q

What is an example of a gene mutation assay in cell culture lines?

A

Mouse lymphoma assay
(MLA) that targets the thymidine kinase
(TK) as selection marker

75
Q

What type of assays are used to detect chromosomal aberrations?

A

chromosomal/cytogenetic assays.

76
Q

What do chromosomal/cytogenetic assays assess?

A

metaphase spreads

77
Q

How do the chromosomal/cytogenetic assays assess metaphase spreads?

A

by using new technology called Fluorescent in situ hybridization
(FISH)

78
Q

What does FISH stand for?

A

Fluorescent in situ hybridization.

79
Q

What is FISH?

A

it assays various chromosomal aberrations.

80
Q

Where can FISH be done?

A

cell culture, plants, animals, and humans.

81
Q

What is genomic DNA isolation used for?

A

for evaluating genomic DNA

82
Q

What is genomic DNA isolation?

A

series of steps to purify DNA from cell.

83
Q

What is needed for gel electrophoresis?

A

determine DNA size, genomic DNA assessing degradation, and cut DNA with restriction enzymes.

84
Q

What is a southern blot?

A

analysis of specific DNA sequences.

85
Q

What do southern blots do?

A

determine if gene is present, copy number of gene, and genotype of genetically modified animals.

86
Q

What does PCR stand for?

A

polymerase chain reaction

87
Q

What does PCR do?

A

amplifies targeted DNA sequences using specially designed primers.

88
Q

PCR can make billions of copies of what?

A

target sequence of DNA in a few hrs.

89
Q

When was PCR first invented?

A

1984

90
Q

What is PCR now used for?

A

cloning, mutagenesis, and analysis of gene expression.

91
Q

What is RNA isolation in gene expression assays?

A

extra caution to prevent degredation of RNA, snapshot of gene expression.

92
Q

What is northern blot?

A

Measures the amount and size of RNAs transcribed and estimates their abundance.

93
Q

What are methods for the Northern blot?

A

runs in agarose denaturing gel, transfer to membrane, probe specific DNA sequence, and probe detection.

94
Q

Why do many assays utilize stable cDNA synthesized from RNA sequence?

A

because a double strand is stronger than a single strand.

95
Q

What does quantitative PCR do?

A

Quantify target gene relative to a reference gene.

96
Q

What does toxicogenomics stand for?

A

genome wide analysis in toxicology.

97
Q

What does genomics mean?

A

the study of genes and their functions.

98
Q

What does transcriptomics mean?

A

The study of mRNA

99
Q

What does Proteomics?

A

The study of proteins.

100
Q

What does metabolomics mean?

A

The study of molecules involved in cellular metabolism.

101
Q

What does glycomics mean?

A

The study of cellular carbohydrates

102
Q

What does lipomics mean?

A

The study of cellular lipids.

103
Q

What does ionomics mean?

A

All mineral nutrients and trace elements of an organism.

104
Q

What includes transcriptomics, proteomics, and metabolomics?

A

Toxicogenomics.

105
Q

Transcriptomics compares what?

A

the genes expressed (mRNA) in an organism that have been exposed to a drug, chemical, or a toxin to those of unexposed organisms.

106
Q

What is the goal of transcriptomics?

A

to identify patterns of gene expressions related to specific chemicals or chemical classes to be used as endpoints in assessing toxicity.

107
Q

What is the end result of microarray transcriptome analysis?

A

differentially expressed gene list related to specific chemical.

108
Q

Why is microarray transcriptome analysis good?

A

if you don’t know molecule targets initially.

109
Q

Why evaluate for genetic damage?

A

lethal mutations, cancer, developmental abnormalities.

Toxicology Final (7 decks)

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