qualifying exam cram Flashcards

1
Q

Can you pass this exam

A

Yes

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2
Q

In the Wang et al paper about how Tip60 depletion protects against myocardial infarction, what mice did they use?

A

mice containing floxed Kat5 alleles, wherein exons 3–11 comprising two-thirds of the Tip60 coding sequence including the chromo and acetyltransferase domains are removed by Cre-recombinase. Mice were B6/SV129 background

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3
Q

In the Wang et al paper about how Tip60 depletion protects against myocardial infarction, how old were the mice used, when was tam given and when was the MI given?

A

10-14 weeks, tam was given day “0” and MI was given 3 days later – they saw about 50% reduction in Tip60 in whole tissue

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4
Q

In wang et al paper where did they get their tissue slices

A

IF staining was from 1mm below suture to apex; qpcr was from 1mm below suture to base

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5
Q

2 ways to verify successful MI

A

cTnT/cTnI in serum (RIP)

Elevated ST segment in EKG being recorded during surgery (Tombstone, also RIP)

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6
Q

which is the base and which is the apex of the heart

A

base is top, apex is bottom, because life is meaningless and scientists are annoying

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7
Q

Left ventricular anteroposterior internal diameter (LVID)

A

You want this to be a certain range becauase you don’t want it to be too big (dilated ventricle) or too small (hypertrophied ventricle)

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8
Q

posterior wall thickness (LVPW)

A

also indication of wall thickness - dilation vs hypertrophy

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9
Q

left ventricular internal area (LVA)

A

area of LV cavity; at systole and diastole, this can be used to calculate fractional area change (FAC) which can give an idea of ventricular function

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10
Q

Fractional Shortening

A

By using the formula: (LVEDD - LVESD / LVEDD) x 100 we get the percentage of size differences of the left ventricle as a parameter of how well the left ventricle is contracting itself and therefore reduces the size during systole. Values > 28% are considered to be normal.

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11
Q

Ejection fraction

A

ejection fraction (EF %) = (end-diastolic volume – end-systolic volume)

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12
Q

can you pass this exam

A

yes

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13
Q

what does ionomycyin do

A

It’s a binder to Ca and Mg and acts as calcium activating - to the extent that it can fucking cause apoptosis

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14
Q

are either acetylation sites on p62 in the consensus sequence you proposed for k464

A

of course not b/c life is hard

K420 and 435 on UBA domain (for noncovalent ub association)

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15
Q

are either acetylation site on ulk1 in the consensus sequence you proposed for k464

A

yes kinda, K606 on ULK1 is RXXXK which is close to KXXXK and honestly i’ll take it at this point
Other is k162

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16
Q

is the ryanodine receptor acetylated

A

thank the fucking lord no

17
Q

la place’s principal

A

T=deltaP*R/wall thickness

In other words, the thicker the wall, the less tension there is (good)

18
Q

did wang et al find anything about apoptosis with depleting tip60

A

Tip60 in the infarcted adult heart is associated with activation of the CM cell-cycle, concomitant with reduction of apoptosis and preservation of cardiac function

19
Q

Is K464 SERCA2A conserved in mammals

A

yes ma’am

20
Q

Does k464 exist in mice

A

yes ma’am thank god <3

21
Q

does serca k464 exist in rats

22
Q

tau

A

exponential decay time constants of calcium

23
Q

bl10%, etc

A

Rise time of the calcium to x% of the peak

24
Q

what is the promoter the CRISPR and GCAMP vectors are behind

A

CAG prmoter – synthetic promoter that strongly drives expression in mammalian models

25
mice used in gorski et al
Studies were conducted in male C57BL/6J mice aged 8 to 10 weeks (weight, 25–30 g) purchased from Jackson Laboratories -- they used a tac model when looking at acetylation and serca in the live animals as well as general sirt1 knockdown -/- this paper also looked at what happens if you induce sirt1 w/b-lap
26
meraviglia et al paper showed saha has what effect on serca
no change of serca2a expression in adult rats- but increased serca activity in rat microsomes when given rat hearts 1-10uM Ca Specifically, SAHA induced a 21% decrease in the time constant tau, as well as a significant reduction in the time to 10%, 50% and 90% of fluorescence signal decay (BL10, BL50, BL90; Figure 3c) (rat hearts). Consistent with this finding, SAHA also affected CM mechanics during the re-lengthening phase, as documented by the significant increase in the maximal rate of re-lengthening (+dl/dtmax, approximately 16%) associated with a decrease in the time to 10%, 50% and 90% of re-lengthening (Figure 3b,d). Conversely, the average diastolic sarcomere length, the fraction of shortening and the maximal rate of shortening were comparable in CTR and CTR+SAHA cardiomyocytes (Figure 3d)
27
What is another possible reason meraviglia et al proposed for the SERCA2A change
possible phospholamban binding site
28
PTip60 by gsk3b
ser86
29
Activated perk
p-perk is pSer980
30
Tip60 role in apoptosis
activation of p53
31
How did the Wang et al paper assess tip60 apoptosis
primarily through tunel staining but also via p21 expression (cell cycle inhibitor)