Determinación de la oferta hídrica en la cuenca aplicando el modelo WEAP

infiltration in all layers are seen. The mucosa shows the heaviest infiltration and also superficialulceration represented by destruction of the epithelial border, m-mucosa sm-submucosa mu-muscle

a)

Figure 5.8 - Obstruction model.

Figures show the anastomotic segment 24 h after resection of the obstruction and anastomosis formation.

Formalin fixed tissue stained with H&E

At 24 h there was still a large gap between the cut edges of the bowel filled with clot, fibrin, oedema and detached inverted muscle. Note the oedematous submucosa and the heavy infiltration of inflammatory cells mainly in the fibrin plug, in the submucosa and around stitches. Original magnification x 25.

m-mucosa sm-submucosa mu-muscle fp-fibrin plug s-stitches.

mucosa submucosa

muscle

mmv

Collagenase I I TIMP I I Gelatinase 1 I and Stromelysin I I are represented diagrammatically. Dots fO ) show intracellular gelatinase,

Figure 5.8. Obstruction model

Figure is a detail from fig 5.8.a.

c) Original magnification x 100

Stitches are seen to be surrounded by large numbers of inflammatory cells 24 h after anastomosis following an obstruction,

Figure 5.9^ Obstruction model.

Figures show anastomotic segment 3 days after resection of the obstructed segment and anastomosis

a)^ Formalin fixed tissue stained with H&E

After 3 days, similarly to the normal healing model, a large oedematous fibrous tissue, heavily infiltrated by inflammatory cells occupied the gap between the bowels ends. At this site mucosal and muscle remnants undergoing degradation are also present. Original magnification x 25

m-mucosa sm-submucosa mu-muscle ft-fibrous tissue.

mucosa submucosa muscl

Collagenase L_J TIMP I— I Gelatinase I— I and Stromelysin I I distribution shown diagrammatically. 0 0 represent intracellular gelatinase which is the dominant feature. Extracellular enzyme is restricted to

the stitch line.

Figure 5.9 - Obstruction model.

Figures 5.9 c to f are ail from the anastomotic segment of the same rabbit 3 days after resection of the obstructed segment and anastomosis. The sections have been stained by indirect immunofluoresce for the MMPs and TIMP-1 including a counterstain for the nuclei which fluoresces red

c) Original magnification x 400

d ) Original magnification x 400

In

c)

the tissue was incubated with antiserum to collagenase and in

d)

with anti TIMP-1. In the mucosa both the enzyme and the inhibitor were

observed as strips of green fluorescence (arrows) surrounding mucosal glands.

Figure 5.9. Obstruction model. Figures show the anastomotic segment 3 days after operation.

e) Original magnification x 400

f) Original magnification x 400

The section in

e)

was stained with antiserum to gelatinase and the section in

f)

was incubated whit antiserum to stromelysin. Both enzymes were immunolocalised as streaks of green fluorescence around the mucosal glands (arrows). Note that the TIMP-1 staining (d) is much less extensive and duller than that of the MMPs.

Figtire 5 J 0 - Obstruction model.

Figures are the anastomoticrsegment 7 days after

resection of the obstruction andformation of anastomosis. a ) Formalin fixedtissue stainec^with H&E

7 days after anastomosis the mucosal continuity is almost restorecfancFa mature fibrous tissue has formedrOedema ancHnflammatory cell infiltration are still seen in the anastomoticregion. Original magnification x 25-

m-mucosa sm-submucosa mu-muscle ft-fibrous tissue s-stitches

mucosa submucosa

muscle

Collagenase I— I TIMP I— I Gelatinase and^tromelysin I— I distribution at the anastomosis. By 7 days MMPs ancfTIMP are only present at the stitch line.

Figure SA 1 - Obstruction model.

Figures show ProxO segment after 24 h of obstruction a)-Formalin fixed tissue stainecfivith H&E

s m

I rn u

In the mucosa on the left side of the photo extensive crypt damage andloss of epithelial border have occurrecf(fig-5J 1 .cr,xt)TOedema, inflammatory cell infiltration ancfhaemorrhage can also be seen throughout the segment especially in the submucosa anchnusde.

m-mucosa sm-submucosa mu-muscle. Original magnification x 40

b)

Mucosa Submucosa Muscde — ► o o 0 o o o Diagram showing-distribution of

Collagenasel— I TIMP I— I Gelatinase I— I ancfStromelysin L-—I in the ProxO segment. Where the epithelial border is damaged^ extracellular enzyme is seen whereas MMPs ancfTIMP are seen in the other layers but only intracellularly (O o y

Figure 5.11. Obstruction model

Figures show the ProxO segment after 24 h of obstruction at the site of most extensive mucosal damage.

c) Original magnification x 100

;

%

d ) Original magnification x 100

Both figures are details of fig 5.11.a. showing the extensive superficial ulceration evidenced by loss of the epithelial border and many crypts . Note the heavy infiltration of inflammatory cells at these sites. This was characteristic of the mucosal damage seen in the most proximal segments of the obstruction model, m-mucosa mm-muscularis mucosae sm-submucosa

Figure 5.11. Obstruction model

Figures show the ProxO segment after 24 h of obstruction e) Original magnification x 1000

A section showing the submucosal layer incubated with antiserum to

gelatinaseA and B. The presence of intracellular gelatinase is demonstrated by green fluorescent granules near the nuclei (counterstained to fluoresce red) of a large number of cells. The elongated nuclei showing granules at one extremity appear to be fibroblast-like cells (thin arrows). The cells with granules

Figure 5.12 - Obstruction model. Proxl segment after 24 h of obstruction

a)

Formalin fixed tissue stained with H&E

Note the extensive damage of the mucosa manifested by total loss of the epithelial border and destruction of crypts. Inflammatory cell infiltration (fig 5.12.c,d), oedema and haemorrhage in all layers can also be seen, m-mucosa sm-submucosa mu-muscle Original magnification x 40

b)

Mucosa

Submucosa Muscle

Collagenase I T: I TIMP d l Gelatinase E S and Stromelysin I I distribution in Proxl segment. Extracellular MMPs dominate in the mucosa whereas only intracellular gelatinase ( 0 ) is observed in the

Figure 5.12. Obstruction model

Figures are magnification of areas from fig. 5.12.a.

c)

Original magnification x 100

Note the extensive infiltration of inflammatory cells in aU layers of the bowel wall especially in the mucosa where many epithelial glands are missing, m-mucosa mm-muscularis mucosae sm-submucosa mu-muscle

d ) Original magnification x 200

. •

Detail of the preceding figure showing loss of the epithelial border of the mucosa, gland destruction and heavy inflammatory cell infiltration, containing both acute and chronic cell types.

Figure 5.13 - Ischaemic model.

Figures show tissue taken 12 h after anastomosis a) Original magnification x 400

Figure shows the mucosa of the ProxO segment incubated with

antiserum to stromelysin. Bands of green fluorescence demonstrate the binding of stromelysin to the ECM (arrows). The nuclei have been counterstained to fluoresce red.

b) Original magnification x 400

T

»

«

Tissue taken from the distal segment incubated with antiserum to TIMP-1. The inhibitor is seen intracellularly surrounding the nuclei which

Figure 5.14 - Ischaemic model.

Figures show the anastomotic segment 24 h after resection and anastomosis.

a ) Formalin fixed tissue stained with H&E

m u

A

s m

24 h after anastomosis the large gap between the cut edges of the bowel was filled with fibrin, haemorrhage, oedema, remnants of mucosal glands and muscle. A large number of inflammatory cells have infiltrated the fibrin plug extending to the submucosa and muscle. Original magnification x 25

m-mucosa sm-submucosa mu-muscle fp-fibrin plug s-stitches

mucosa submucosa

muscle-

Diagrammatic representation of

Collagenase ^ 0 TIMP I

I

Gelatinase and Stromelysin

I

I

at the anastomosis. Intracellular gelatinase fO O ) dominates accompanied by extracellular MMPs restricted to the mucosa.

Figure 5.15 - Ischaemic model.

Anastomotic segment 3 days after resection and anastomosis.

a ) Formalin fixed tissue stained with H&E

'4'-

s m

m u

3 days after anastomosis a fibrous tissue fills the gap between the cut bowel ends. It is infiltrated by many inflammatory cells and also contains remnants of mucosal glands and muscle. Original magnification x 25

m-mucosa sm-submucosa mu-muscle ft-fibrous tissue

mucosa submucosa

muscle

Collagenase EZ] TIMP I I Gelatinase EUl and Stromelysin I— I distribution shown diagrammatically at the anastomosis. Both intra-

CHAPTER 6

In document Modelación hidrológica de escenarios futuros de caudales medios con la aplicación del modelo hidrológico weap en la cuenca del rio Ilave Puno (página 68-80)