bioz476: opioids, genetics & personalized medicine

According to the lecture notes, how many American adults report experiencing "a lot of pain"?

23.4 million adults, which represents $10.3BLABLA_PERCENT$ of the adult population.

What is pain often designated as in healthcare assessment settings?

The Fifth Vital Sign.

What unique pharmacological property differentiates opioids from other pain relief medications?

Opioids have no "ceiling effect," meaning there is no maximum limit to their analgesic effects.

Which class of pain medications takes weeks to produce an effect and does not work well for pain relief?

Antidepressants (used for neuropathic pain, chronic daily headaches, fibromyalgia, and chronic low back pain).

Which brain region is stimulated by opioids to release dopamine?

The ventral tegmental area (VTA).

Dopamine release in which two brain structures causes the feeling of euphoria upon opioid administration?

The nucleus accumbens (Nac) and the prefrontal cortex (PFC).

What happens to dopamine receptors following repeated drug administration?

Receptor downregulation occurs, resulting in drug users having fewer available dopamine $D_2$ receptors.

What physical and psychological symptoms are listed as key signs of opioid withdrawal?

Anxiety, insomnia, rhinorrhea (runny nose), diarrhea, emesis, mydriasis, piloerection ("cold turkey"), restless leg syndrome ("kicking the habit"), and tachycardia.

What was the estimated annual economic burden of the opioid crisis in the United States according to the CDC?

$78.5\text{ billion}$ per year, covering healthcare, lost productivity, addiction treatment, and criminal justice involvement.

What percentage of new heroin users report abusing prescription opioid pain medications prior to using heroin?

Approximately 4 out of 5 people ($80\%$).

What percentage of patients prescribed opioids for chronic pain develop an opioid use disorder?

Between 8 and 12 percent.

What is the full name of the ANKK1 gene and what protein does it encode?

Ankyrin repeat and kinase domain containing 1, which encodes the protein kinase PKK2 involved in brain dopamine synthesis.

Which specific Single Nucleotide Polymorphism (SNP) in the ANKK1 gene is analyzed in the laboratory experiment?

SNP rs1800497.

How does the ANKK1 gene influence dopamine receptor activity?

ANKK1 is located adjacent to the $D_2$ Dopamine Receptor gene (DRD2) and can influence DRD2 expression.

What is the expected size of the ANKK1 PCR product before restriction enzyme digestion?

532 bp.

What fragment sizes are produced after TaqI restriction digest of the ANKK1 PCR product for an individual with the CC genotype?

284 bp and 248 bp fragments.

What band sizes appear on a gel after TaqI restriction digest for an individual with the CT genotype at SNP rs1800497?

532 bp, 284 bp, and 248 bp fragments.

Why does the TT genotype remain at a single 532 bp band after TaqI digestion?

The T allele lacks the TaqI restriction enzyme recognition site, preventing digestion.

How do host bacteria protect their own genomic DNA from being cleaved by their endogenous restriction enzymes?

Bacteria methylate their own DNA.

Which bacterial species produces the EcoRI restriction enzyme, and what type of overhang does it generate?

Escherichia coli; it produces a $5'$ overhang.

Which bacterial species produces the PstI restriction enzyme, and what type of overhang does it generate?

Providencia stuartii; it produces a $3'$ overhang.

How many heroin-dependent patients were included in the study's case group, and what criteria were used for diagnosis?

69 unrelated, heroin-dependent patients diagnosed based on Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-5) criteria.

What are the three main components of a DNA loading buffer and their functions?

Colored dye (tracks sample migration), thickening agent such as sucrose/glycerol (adds density so samples sink into wells), and EDTA (binds $Mg^{2+}$ cations to halt nuclease activity).

Which tracking dye is used in the loading buffer for the ANKK1 lab experiment?

Orange G.

What DNA-intercalating dye is used to visualize bands under UV light in agarose gel electrophoresis, and what safety gear is required?

Ethidium Bromide; a lab coat and gloves must be worn because it is carcinogenic.

What p-value threshold determines statistical significance when analyzing genotype data using the Chi-Square test?

$p < 0.05$.

What population-level genomic method is utilized to uncover unknown genes associated with polygenic diseases or traits?

Genome-Wide Association Studies (GWAS).