TABLE XIV-RESULTS OF EMA IMMUNOSTAINING IN PERITONEAL

FLUID CELL BLOCKS ( N=49)

Histopathological
Diagnosis on cell
block

Reactive Mesothelial
Cells (n=25)

Malignant Cells
(N=24)

Serial
number
of
Cases

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

EMA
Percentage of immunoreactive cellls
No
<10%
10-50%
>50%
cells
(+)
(++)
Cells
(+++)

No stain
(0)

Mild
(+)

Intensity of staining
Moderate
Strong
IHC Grade (sum of
(++)
(+++)
percentage and intensity
of staining

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
+++
+++
++
++
+++

++
++
++
++
++

++
+++
+++
++

+++
+++
++
++

+++
+++
++
++
+++
++
++
+++
+++
++

++
++
++
++
++
++
++
++
++
++

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
5
5
4
4
5
5
6
5
4
0
5
5
4
4
5
4
4
5
5
4

21
22
23
24

+++
+++
++
++

++
++
++
++

5
5
4
4

Table XIV shows the percentage and intensity of immunoreactive cells for EMA
in peritoneal fluid cell blocks. EMA positivity was noted as cytoplasmic and
membranous staining. As shown in the table, percentage of immunoreactive
cells were scored as - no cells stained = 0, < 10% cells stained=1, 10-50% cells
stained = 2 and > 50% cells stained= 3.The intensity of staining was scored as no cells stained=0, mild intensity of staining=1, moderate intensity of
staining=2 and strong intensity of staining=3.The IHC grade was calculated as
the sum of scores of percentage of immunoreactive cells and the score of
intensity of staining. IHC grade > 4 was taken as positive. The cases of
reactive mesothelial hyperplasia were 25 in number. Amongst the reactive
mesothelial cells, none of the cases(0%) showed immunoreactivity for EMA.
Amongst the malignant cases, 22 cases (91.6%)showed positivity for EMA .One
case (4.1%)showed IHC grade 6 while 11 cases (45.8%)showed IHC grade 5
and 10 cases(41.6%) showed IHC grade 4. The two malignant cases that were
negative for EMA were
squamous cell carcinoma.

later diagnosed as hepatocellular carcinoma and

TABLE XV- RESULTS OF CALRETININ IMMUNOSTAINING IN

PERITONEAL FLUID CELL BLOCKS (N=49)
Histopathological
Diagnosis on cell
block

Serial
number
Of cases

CALRETININ

No
cases
(0)

Reactive
Mesothelial Cells
(n=25)

Malignant Cells
(N=24)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18

Percentage of immunoreactive cells

<10% (+)
10-50%
>50%
(++)
Cells
(+++)

++
++
++
++
++
++

Strong
(+++)

Intensity of staining
Moderate
Mild
No
(++)
(+)
stain
(0)

++
++
+++
++
++
++
++
++
++
++

+++
++
++
++
++
++
++
++
++
++
++
++
++

+++
++
++
++
++
++
++
++
++

+
++
++
++
++
++

++
++
++
++
++
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

IHC Grade
(sum of
percentage
and intensity
of staining

4
4
5
4
4
4
5
4
4
4
5
4
4
4
4
4
4
4
4
2
4
4
4
4
4
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

19
20
21
22
23
24

0
0
0
0
0
0

0
0
0
0
0
0

Table XV shows the percentage and intensity of immunoreactive cells for

Calretinin in peritoneal fluid cell blocks. Calretinin positivity was noted as
nuclear and and cytoplasmic staining. As shown in the table, percentage of
immunoreactive cells were scored as - no cells stained = 0, < 10% cells
stained=1, 10-50% cells stained = 2 and > 50% cells stained= 3.The intensity of
staining was scored as - no cells stained=0, mild intensity of staining=1,
moderate intensity of staining=2 and strong intensity of staining=3.The IHC
grade was calculated as the sum of scores of percentage of immunoreactive cells
and the score of intensity of staining. IHC grade > 4 was taken as positive.
Amongst the 25 cases with reactive mesothelial hyperplasia, 24 cases(96%)
showed immunoreactivity for Calretinin.One case did not show positivity and
this had mainly neutrophilic cells with very occasional mesothelial cells that
showed only mild staining with Calretinin resulting in IHC grade 2.Therefore it
was not considered as positive. Three cases (16%) showed IHC grade 5 while
21 (84%) cases showed a IHC grade 4.
Amongst the 24 malignant cases, all cases (100%) were negative

for

calretinin. Clusters of mesothelial cells present along with adenocarcinoma cells

did show positivity for calretinin .However the number of such cells was less
than 5% and they were below IHC grade 4 and were considered negative.