Experience with a new device for pathological assessment of colonic endoscopic submucosal dissection.

Endoscopic submucosal dissection (ESD) is gaining popularity worldwide in the treatment of neoplastic lesions of the gastrointestinal tract. However, the experience in Western countries is quite limited and restricted to large or academic centers. Besides, this approach requires an optimal pathological assessment. The aim of this study was to report our experience with colonic ESD using a new device that allows complete handling of the resected specimens and especially of lateral margins, for pathological analysis. In a 1-year period, 14 patients (6 men, 8 women, age range 50-82 years) underwent colonic ESD in a non-academic hospital. The endoscopic procedure was carried out successfully en bloc in more than 90 % of cases. Perforation requiring surgery occurred in one patient (7 %). Pathological assessment with the new device allowed entire and complete examination of both the deep and lateral margins of the excised specimens. Colonic ESD is a viable option for non-surgical treatment of large bowel lesions even in relatively small centers and in non-academic settings. The new device allows good handling of the specimens, and it seems to be useful for the entire examination of the resection margins.

SEM corrosion-casts study of the microcirculation of the flat bones in the rat.

BACKGROUND: Little is known about the organization of microcirculation in flat bones in comparison with long bones. This study, therefore, helps us to determine the design of this vascular system in flat bones in relation to their structure and function. METHODS: The organization of microvasculature in parietal, scapula, and ileum bones of 15 young sexually mature rats, aged 6-7 weeks, was studied by light and scanning electron microscopy (SEM) from vascular corrosion cast (vcc), a resin-cast obtained material. RESULTS: Our observations show that the pattern of the microcirculation in flat bones is different in the thick and thin parts of such bones. Where the bone is thinner than 0.4 mm, only periosteal and dural network exist. Larger vessels which do not form a real network connect the two tables of the bones in these regions. In thicker areas, the organization of the microvasculature is similar to that in long bones, with distinct periosteal, cortical and bone marrow networks. Moreover, in different bones, outer networks show slightly different characteristics according to the different adjacent structures (dura mater, muscles etc.). Different types of vessels were recognized by comparing their different diameter, course and endothelial imprints. CONCLUSIONS: The microvascular patterns of the flat bones are strongly influenced by the bone thickness. The different microvascular systems can interact both with the bone modelling and remodeling and with the variable metabolic needs, modifying the microvascular pattern and the blood flow. This is even more important in view of the reciprocal influence of the different networks within the same bone.

Hepatic microcirculation as a morpho-functional basis for the metabolic zonation in normal and pathological rat liver.

The hepatic microcirculation is well known as a fundamental component of the liver structure, deeply involved in the zonal organization of the acinar structure. In cirrhosis, the microvascular tree shows dramatic changes that would heavily influence the development of the disease. When the cirrhosis becomes evident the result is a progressive organ failure, also in presence of only moderately decreased hepatocyte volume. The aim of this research was to compare the role of microcirculation of the hepatic zonation in normal and cirrhotic livers. Cirrhosis was experimentally induced in 36 rats following a controlled intragastric CCl4 administration. Cirrhotic and control normal livers were processed for routine light microscopy, histoenzimology, and scanning electron microscopy vascular corrosion cast. Control livers showed normal hepatic structure and microvascularization; enzymatic activities were constantly and normally distributed. In CCl4-treated animals LM showed a characteristic micronodular cirrhosis in all livers. Vascular corrosion casts under the scanning electron microscope displayed a progressive reduction of the distance between pre- and post-sinusoidal vessels and the presence of newly formed perinodular plexus. The histoenzymatic analysis demonstrated the loss of zonation in the cirrhotic parenchyma. Moreover, the sinusoid/hepatocyte ratio was significantly reduced, because of the presence of two or more hepatocyte thick laminae during the scarring development. The altered microcirculation in cirrhosis also changed the normal acinous metabolic gradient. The histoenzymatic study revealed a zonal rearrangement of the cirrhotic liver metabolic activity, that leads to a progressive hepatic failure. These data confirm the fundamental importance of the normal relationship between the hepatocyte laminae and the sinusoids for the preservation of a normal zonation which represents the basis for a normal liver function.

Microcirculation of the extrahepatic biliary tree: a scanning electron microscopy study of corrosion casts.

The microvascular arrangement of the extrahepatic biliary tree of the rat was studied by light microscopy (LM) and scanning electron microscopy (SEM) of vascular corrosion casts. The plexus that encircles the lumen of the common bile duct, observed by LM, showed a network of vessels of different diameter situated under the epithelium in the lamina propria. Parallel SEM observations of the same structure demonstrated the presence of 2 main vascular layers: an outer arterial and venous layer, corresponding to the larger vessels seen by LM, and a richer inner capillary layer just under the epithelium. On the luminal part of the corrosion casts, there were many round avascular empty pits that corresponded to the presence of small acinar glands distributed along the epithelium of the common bile duct. The rich subepithelial capillary network present in the rat, an animal without a gallbladder, may play an important role in the reabsorption of water and solutes from bile. Moreover, in pathological conditions (e.g. portal hypertension), liver blood flow may take a preferential collateral route through the intrahepatic peribiliary plexus into the relatively large diameter vessels of the extrahepatic peribiliary plexus because of the continuity of the 2 plexi.

Intrinsic innervation of the rat knee joint articular capsule and ligaments.

In spite of the practical importance of having a detailed knowledge of knee joint innervation to understand the pathophysiologic aspects, little information is now available concerning the density and pattern of the nerve fibres which are distributed to it. The present study has been designed to investigate the density and distribution of nerve fibres and receptor corpuscles in the knee joint articular capsule, cruciate and collateral ligaments in the rat, using the acetylcholinesterase (AChE) histochemical in toto staining technique. The investigation was performed on male Wistar rats of 3 months of age, some of which had been treated with capsaicin to deplete their afferent 'C' fibres of their content of neuropeptides. AChE-positive nerve fibres and different types of receptor corpuscle endings were found within articular capsule and ligaments. The highest density of AChE-positive nerve fibres was noticeable in the fibular collateral ligament followed by the tibial collateral ligament, the posterior cruciate ligament, the anterior cruciate ligament and the articular capsule. In the articular capsule the number of type I endings was higher than in the ligaments. The opposite is true for the other type of receptor corpuscles found as well as for nerve endings. Capsaicin treatment significantly reduced the density of AChE-positive nerve fibres in knee joint ligaments but did not affect nerve fibres in the articular capsule. Moreover, it caused the disappearance of some kind of receptor corpuscles within the collateral and cruciate ligaments. The above data collectively suggest that the AChE in toto staining technique may represent a good method for investigating joint innervation and that a significant percentage of nerve fibres supplying knee joint ligaments is represented by C fibre afferents.

A scanning electron microscopy morphometric study of the rabbit peritoneal surface.

Rabbit peritoneum was studied by SEM to obtain information and statistically meaningful morphometric data of different sites of visceral and parietal peritoneum and to verify the existence of "stomata." Samples were fixed by intraperitoneal infusion of glutaraldehyde, and were photographed by SEM under standard conditions. Morphometric data were obtained by Kontron MOP Videoplan. Variable cell surface patterns were present even within limited areas; however, "stomata" were not observed. The heterogeneity of data obtained can be related to the dynamism of mesothelial cell activity and to the different motilities of the underlying organs.

A three-dimensional study of the morphology and topography of pericytes in the microvascular bed of skeletal muscle.

Digested tissue specimens and corrosion casts of rat soleus and tibialis anterior muscles were employed for this Scanning Electron Microscopy (SEM) study. The shape, morphology, and position of pericytes were compared to corresponding imprints on the cast surfaces. Pericytes, observed in digested tissue specimens, showed a typical morphological pattern: a central body with two primary processes that run along the capillary in opposite directions. From these primary processes, secondary ones arise and often encircle the vessel almost completely. On the surface of corrosion casts, roundish imprints were found in the microvascular tree at the same level where digested tissue specimens showed the presence of pericyte bodies. Along and around the cast surface, shallow grooves reproduced the course of the primary and secondary processes. The peculiar tridimensional arrangement of pericytes at the level of capillary bifurcations underlines their role in red cell flow regulation. However, if the mechanical linkage of the pericytes to the endothelium and their contractability is taken into account, additional roles of these perivascular cells may be hypothesized.

Microcorrosion casting in normal and pathological biliary tree morphology.

The organization of the intrahepatic biliary tree was studied in three dimensions by scanning electron microscopic (SEM) corrosion casts, in normal and cholestatic rat liver. In the normal liver the observation revealed the features of the biliary passages from the bile canaliculi to the canaliculo-ductular junction, to the ductules and the bile ducts, confirming previous SEM observations. In cholestatic liver, the modifications and the proliferation of bile ductules appear clearly. Resin flow from canalicular to sinusoidal network was never observed. The method was found to be very useful in the evaluation of the architecture of the intrahepatic biliary tree, under normal as well as under pathological conditions.

Surface morphology of the human pleura. A scanning electron microscopic study.

The superficial morphology of the pleura has been observed in humans by scanning electron microscopy (SEM). Pleural samples from the visceral, mediastinal, diaphragmatic and costal normal human pleura were observed, and a thorough morphometric study was performed. The most evident feature was the ubiquitous presence of microvilli and micropores. The secretion and absorption activities therefore should not be regarded as accomplished by different topographic zones of the pleura. Discontinuities and clefts were observed at the level of cellular junctions; absorption of high weight molecules and cells from the pleural space may be facilitated at this level, but the structures which were observed in humans may not be considered "stomata," according to the definition of such morphologic units as we give in the present study. Cilia and blebs, described in the experimental animal or in other mesothelia, were not seen in the human pleura.

Measurements of cross-sectional area of collagen structures (knee ligaments) by means of an optical method.

Measurement methods of cross-sectional area of collagen tissues and structures, so far chosen, are critically reviewed. A new optical method, founded on section outline plotting, by which it is easy to compute the cross-sectional area, is suggested and shown in particular. Experimental results, achieved by applying the method to the measurement of bovine knee ligament sections, are reported; these results are compared with those taken from samples coming from the same ligaments.

Microcorrosion casts in the microcirculation of skeletal muscle.

The authors review the contribution of microcorrosion cast studies towards clarifying the structure of skeletal muscle microcirculation. Former studies performed on naturally contracted muscles show the presence of a primary and a secondary arterial network and a capillary network. At the level of the capillary network pericyte imprints are present. Muscles characterized by different types of metabolism show different features of the capillary pattern. Other authors have affirmed that the extended muscle is characterized by long and straight capillaries, while the contracted one features clusters of vessels all around a muscle fiber. The authors have made the present observations in order to determine how the capillary pattern of muscles with different metabolism is modified by extension and shortening of the muscle belly. The capillary pattern observed appears very similar to that observed in former studies. The differences between the oxidative and the glycolytic muscle are evident in every condition of the muscle belly. These data confirm the theory that there is a permanent endogenous difference in microcirculation between oxidative and glycolytic muscle, determined by muscle fiber metabolism.

An S.E.M. corrosion cast study on pericyte localization and role in microcirculation of skeletal muscle.

The authors used the microcorrosion cast technique to study the localization and the role of pericytes in the microcirculation of skeletal muscle. Both fast twitch muscles, the tibialis anterior and the pectineus muscles, and a slow twitch one, the soleus muscle, were studied, because of morphological differences of their capillary networks. The observation of peculiar imprints on the surface of vascular casts, strategically sited in the microvascular bed, were related to the presence of pericytes apposed to the endothelial lining. The role of pericytes in equalizing the local blood flow to metabolic needs and in maintaining capillary blood flow despite pinching of vessels is discussed.

Human knee ligaments: mechanical tests and ultrastructural observations.

The authors carried out mechanical tests on the collateral and cruciate ligaments of the human knee, using two different types of apparatus. They report and discuss the results of these tests relative to loads, stiffness and deformations. They also describe the electron microscopic findings in samples taken from above and below the macroscopic rupture. These observations are compared with those made in samples from unstressed ligaments.

A scanning and transmission electron microscopic study of experimental extrahepatic cholestasis in the rat.

The rat liver after extrahepatic biliary obstruction was studied by SEM and TEM in correlation with basic histochemical techniques. Cholestasis was verified by serological methods. The biochemical data (increase in serum bilirubin values, a gradual lowering of the albumin fraction), in agreement with the ultrastructural results of a sparse RER, suggested a gradual decrease of the protein synthetic activity of the hepatocyte. SEM and TEM revealed numerous fat-storing cells, closely associated with patches of connective fibrils in the subendothelial spaces. Further ultrastructural observations demonstrated: a) a proliferation of the intrahepatic biliary tree (ductular proliferation, including newly formed ducts with sacculation and diverticuli); b) an increased number of canaliculo-ductular junctions and, c) an increase in the length of the bile canalicular network due to its tortuous course, pocketing and side branching. The occurrence of an intact cytoplasmic barrier separating the bile canalicular lumen from the Disse's space together with the results obtained by retrograde infusion of ferritin into the biliary tree suggested that the regurgitation pathway by ductular reabsorption and by transhepatocytic transport is the best documented and most acceptable, at least in the rat.

The glycocalyx of the epithelial cells of the colon, observed in normal and ulcerous colitic conditions.

Biopsies of subjects affected by ulcerous colitis were stained with ruthenium Red. Alterations of the cellular coat and glycocalyx of the epithelial cells of the colon were identified.