Journal Club Figure Facts Worksheet

Spring 2021

Journal club focuses on the figures presented in the paper. The instructions and
questions below walk you through analyzing this research paper.

Come to class expecting to present and discuss the figures and how they support the
research in the article. During class, each team will work together to present a section of
the paper (intro, conclusions) or a figure, using the Figure Facts format and discuss how
the paper as a whole relates to our research projects.

Instructions:
1. List the name of the science article, date published, last author, and affiliation of
the last author (where they work). Use RED TEXT
a. SARS-CoV-2 infection of the placenta, August 17, 2020, Department of
Obstetrics, Gynecology, and Reproductive Sciences at Yale University
School of Medicine

2. Skim the entire article to understand its structure and basic premise.

3. Read the abstract and introduction more slowly.

a. In your own words, what is the question(s) the authors are trying to
answer here? (1-2 sentences) Use RED TEXT
The authors are trying to determine two things: if SARS-CoV-2 of the
placenta can ultimately infect or impact the fetus otherwise, and if a
SARS-CoV-2 infection is able to cause severe morbidity in women who
are pregnant and become infected.

b. In your own words, what do you think is their hypothesis in this research
project? (1-2 sentences) Use RED TEXT
They directly stated their hypothesis, but it is that they believe that the way
SARS-CoV-2 uses the ACE2 receptors in the placenta could cause an
earlier showing of hyptertensive disorders during pregnancy (in women
who would already likely have these disorders) and could also worsen the
condition because of the way it infects the placenta.
 c. Summarize key points from the introduction that are important to know to
understand the research in the paper. Use RED TEXT
i. Mom was a healthy 35 y/o Asian American woman who got sick
from her husband, who was exposed at work. She began exhibiting
mild symptoms 10 days before being admitted to the hospital,
worsening in the 4 days before being admitted (acute symptoms).
1. Day she was admitted - vaginal bleeding, abdominal pain,
high blood pressure, not fevered
2. Was 22 weeks along
ii. We still don’t know the risks and effects of SARS-CoV-2 on
pregnant women.
iii. Only research prior was a small cohort of women who were sick
when pregnant in 3rd trimester - no adverse effects
iv. Hypertensive disorders of pregnancy (HDP) complicate 2-8% of
pregnancies (aka rather rare to begin with) even more rarely occur
in the second trimester (normally a first trimester issue).
1. Limited case reports regarding this and COVID-19
v. HDP exhibited in this case was clearly different from that of typical
preeclampsia

4. Study each figure and fill in the table below.

a. Read each figure legend

b. For each figure, find and read the associated text in the main body of the
article (typically, but not always, this is in the results section).

i. Consider skimming the methods section to better understand the

assays and techniques used.

ii. Consider reviewing the text in the conclusion section to better

understand the takeaways from each figure and experiment.

c. Use your own words to fill in the table below to describe each figure.
i. Only use terms that you understand, can define, and explain; Use
web searches, office hours, or other sources for help. Do not copy
or quote from the article.

ii. Don’t write full sentences in the table, just be clear and concise.
You need to be able to present this and answer questions on it, so
make sure you understand everything you write.
 d. Add more rows or delete rows in the table as needed. The table should
work for YOU. This is YOUR summary of the figures and what the authors
discovered. Think of it as a “cheat sheet.” So, make this “cheat sheet”
work best for you!

i. Figures with multiple images panels may be best explained split

over different rows if they represent different experiments.(ex. 1a
on a row and 1 b on another row) However, if the images are part
of one experiment, keep them on one row (ex. controls and
experimental should be kept together). Use RED TEXT.

5. Once you have completed the Table below, answer the following questions:

a. What are the major conclusions of the article? Use RED TEXT
i. SARS-CoV-2 can cross the placental barrier, and when it does,
there can be drastic issues impacting both maternal and fetal health
and outcomes. High titers of SARS-CoV-2 in the placenta were
found, as well as SARS-CoV-2 RNA in parts of the placenta (as
seen in figures 3 and 4).
b. How does the research in this work relate to the bigger picture (to society,
our community, the world, etc)? Use RED TEXT
i. This helps us to understand how big of an impact SARS-CoV-2 can
have on pregnant mothers and fetuses. More research can be done
to determine if placental infection is detrimental to maternal health,
and if it could ultimately infect the fetus. Ultimately, this can also
help us eventually determine necessary treatment options for
pregnant women, as well as preventative steps that can be taken.
Overall, this is something that is still vastly unknown and can be a
large stressor for pregnant women during this pandemic.

Table 1. Overview of Journal Club Paper

Figur What are we looking If it’s part of an The “take home” How does
e at in this figure? experiment: message of the the work in
[Ex. a mouse embryo what are the figure is… this figure
stained to show actin controls (if lead into the
protein, bar graph used)? (use your own work in the
of…] language, do not next figure?
 What are the copy from text or
Technique/tools/type variables or use science If it’s the last
of imaging used in what is being jargon). figure, what
the figure. compared? should be
Summarize. done next?
If not an
experiment,
what is its role
in the article?

Fig 1 A - This is a timeline A - Is A - the biggest These two

highlighting the comparing take away from figures tie
patient’s symptoms, what happened this is that the well
procedures, and each day to the patient tested together. It
overall outcomes patient, serves positive for is
over a 15 day period. as a basis to SARS-Cov-2 on foundational
orient the day 10, and that to the rest of
B - This part of the reader to how is when the paper. It
figure focuses in far apart the everything went ties into the
again on the worst events downhill. That is next figure
day of the infection, happened. the day the as the next
day 10, and follows a pregnancy was figure
74 hour period from B - compares terminated, the discusses
when the initial labs three different patient was postoperativ
were completed. statistics/count transferred to the e results
s against each ICU, and from qRT-
other: platelet exhibited the PCR tests,
count, worst symptoms. which is
hematocrit Could be chronologic
(RBC in the considered the ally after the
blood), and “peak” of the second part
fibrogen (how infection of Figure 1.
well the B - the platelet
coagulation count (green
factor is line) is shown to
working) be somewhat
LOW until hour
21, and then
swiftly rose over
the next 50 hours
 or so, putting her
back into normal
levels. The
hematocrit levels
remained rather
normal
throughout the
whole period
after a slight
drop. The
fibrinogen levels
skyrocketed from
the point of blood
product
administration at
hour 7 to hour
21, whenever the
D&E was
performed. This
leveled off a bit
afterward but
may have
continued to rise
to dangerous
levels if a D&E
was not
performed. (this
is hard to
determine
because we
don’t know when
the patient
needed
recessatated).

Fig 2 A - This figure is A - Is A - The only This figure is

showing all of the comparing the places that came a bit of an
different samples that different back positive for overview
were taken from the locations that SARS-CoV-2 that goes
patient (maternal were tested for were the more in
 samples) and the positivity, and placenta, depth in
fetus, as well as the compared them umbilical cord, figure 3.
placenta and via virus titer maternal saliva, This talks
umbilical cord. levels, N1 and and maternal about the
N2 primer urine. The nasal places that
B - This is a probe Ct and oral swabs were
phylogenetic tree of values, and RP both had non positive and
the different SARS- primer and detectible levels the strains,
CoV-2 genomes probe Ct of virus probes, whereas
based on location, values. which was Figure 3
and is categorized by notable given the shows the
the different B - As mother had just histopatholo
substitutions (genetic mentioned tested positive gy of the
divergence). It before, this is earlier on that placenta.
includes the comparing the same day. The
sequence found in different strains Almost every placenta
the placenta of SARS-CoV- other sample had was the
compared to the 2 to the Wuhan at least one location with
Wuhan sequence (at isolate. This is undetectable the highest
least it is assumed in the article to level. titers.
because it was never emphasize how
mentioned). All of the closely related B - The SARS-
information was the SARS- CoV-2 isolate
taken from GISAID. CoV-2 sample from the placenta
taken from the is only 0.0003
placenta is to different from the
other isolates original isolate,
from around meaning that
the world, as there are only 3
well as to the nucleotide
original strain. substitutions in
the entire
genome
(A28881A,
G28882A, and
G28883C).

Fig 3 A - C - This is a A - C - These A - C - Immune This figure

stained section of three images cells have taken includes
placenta showing serve over various images of
histiocytic somewhat as a open spaces in different
intervillositis at threecontrol for what the placenta. areas of the
different to compare the This indicates placenta
magnifications. The other placental that there was a that are
star is showing stains. They need for an stained for
where immune cells are showing immune various this,
have filled in that staining that is response in the and the next
space. not directly area, but it could figure is
SARS-CoV-2 have been more
D & E - Another stain related, but is unrelated to images of a
of the placenta that is of the same SARS-CoV-2 placenta,
staining for CD68 placenta. (instead from although
presence at two histiocytic they were
different D & E - These intervillositis, taken with
magnifications. images are which is a an electron
Presence is indicated compared to pregnancy microscope
by brown coloring. the first three disorder). There and are of
images, but is could possibly be different
F - Similar to D & E, specifically a connection placental
but staining for CD3, staining for between SARS- cell types.
which is a marker of CD68 marker. CoV-2 and an Different
T cells. CD68 is a very increased risk of parts of this
well known and histiocytic figure show
G & H - Staining for commonly intervillositis. SARS-CoV-
SARS-CoV-2 spike used marker 2 in the
protein presence in protein. D & E - These placenta,
the placenta at two are showing that and the next
magnifications. F - Similar to there are CD68 figure does
image D, but macrophages as well.
I - In placenta tissue with CD3 present in the
analysis for SARS- marker area, and they
CoV-2 RNA, make up a
indicated by dark G & H - Images majority of the
red/brown staining. similar to D & infiltrate in the
E, but instead area.
staining for
SARS-CoV-2 F - Similarly to D
spike proteins. & E, this stain is
Again, showing that
compared to A there are T
- C. It is meant lymphocytes
to show present at this
progression placental
and that the location. This is
placenta was in determined
fact infected. through the use
of CD3, which is
I - This image a marker protein.
is a bit more in
depth and is in G & H - Similarly
tissue, and is to the rest of the
meant to show images, this is
the progression staining for a
of infection specific protein,
compared to but in the case of
the other these images, is
images. SARS- staining for
CoV-2 RNA is SARS-CoV-2
clearly present spike proteins.
here. This is probably
because the
spike proteins
are necessary for
attachment. The
images show
that there is a
highly
concentrated
localization of
SARS-CoV-2
spike protein in
syncytiotrophobl
ast cells.

I - This stain is
not testing for
proteins, but
RNA. There is a
strong positive
(red/brown) stain
 in this section of
tissue, which
indicates that
there is a large
amount of
SARS-CoV-2
viral RNA in the
tissue. This goes
to prove that the
placenta was in
fact infected.

Fig 4 A - A labeled A - This image A - There really This is the

schematic of serves as a isn’t a last figure,
placental tissue that reference for complicated take but it would
contains the same the rest of the home message be
genetic material as figure, because to this, other than interesting
the fetus. it shows a color to show that the to look at
coded and samples of the tissue
B & C - Two different labeled image placenta were samples
cell types, to compare taken from an from the
cytotrophoblasts and back to. area (chorionic umbilical
syncytiotrophoblasts villi) that contains cord in
in placental, one of B & C - These the same genetic comparison
which is an enlarged are almost a material of the to the
image (C) reference fetus. This could placenta.
photo for D. have been done Even though
D - Image of the These show to show that the titers
cytoplasm of a two different further infection were overall
cytotrophoblast cell types, one would have lower, they
containing labeled of which is the impacted fetal were still
virus particles focus of the tissues in utero. relatively
next image. high.
E - H - Fibroblast Image C also B & C - B serves Additionally,
cells with labeled shows viral as a bigger had this
virus particles in particle picture (literally infection
them. Particles are invasion. and figuratively) progressed
indicated by arrows. before zooming but reached
At several different D - This is one into to show the the same
scales/magnifications of the cell types viral particles outcome,
 shown in found in fetal tissue
I - An endothelial cell images B and syncytiotrophobl samples
with budding viral C. It is here to asts (barrier would also
particles present. show that virus tissue cells). be
particles have There are 4 interesting
infiltrated one distinct places to look at.
of the different where viral Would they
placental cell particles have have
types. infiltrated the appeared
cells. similarly?
E - H - Similar
to D, this is a D - Again, this is
different showing that
specific cell. It there are several
serves the different
same purpose locations within
syncytiotrophobl
I - Similar to D - asts (in this case
H, this is yet the cytoplasm)
another cell where there are
type that is viral particles.
showing virus This emphasizes
particles, but that viral
this is particles are
demonstrating present.
how they are
budding E - H - Three
(showing that different
the virus was magnifications
actively showing that
replicating prior virus particles
to obtaining the have also
sample). infiltrated
fibroblasts, which
is a connective
tissue cell type,
thus showing the
particles are
found further into
the cell.
I - Similar to the
other images,
this is showing
how there are
viral particles
also found within
endothelial cells
(blood vessel
wall cells), which
could mean that
the viral particles
were able to
circulate through
the bloodstream.

OVERALL -
these images
show that there
were viral
particles found
throughout
placental tissues,
including from
the outermost
part to the
innermost, which
could suggest
how widespread
and serious this
infection was.