Porcine coronary arteries: immunohistochemical profile of TNF-alpha, IL-1beta, TGF-beta1 and ICAM-1.

BACKGROUND: In our study we used immunohistochemical technique to demonstrate the presence of the cytokines tumour necrosis factor alpha (TNF-α), interleukin 1beta (IL-1ß), transforming growth factor beta1 (TGF-ß1) and intercellular adhesion molecule-1 (ICAM-1) in porcine coronaries even in physiological conditions. MATERIALS AND METHODS: Inflammatory cytokines are polypeptide mediators which act as a communication signal between immune system cells and other types of cellsin different organs and tissues, both in human and pig coronary circulation. RESULTS: Our results show that pro-inflammatory cytokines TNF-α, IL-1ß, TGF-ß1 and ICAM-1 are also present in the medium tunica of the coronary arteries under physiological conditions. These results may be compared with those found in coronary atherosclerosis, where the increase in TNF-α has a dramatic effect on the function of the left ventricle, and the high value of IL-1 correlates directly with the extent of myocardial necrosis. In our study we observe the damage and activation of endothelial cells; this induces endothelial dysfunction by accumulation and oxidation of low density lipoproteins (LDL). The formation of oxidized LDL could play a central role in the amplification of the inflammatory response causing an increased expression of pro-inflammatory cytokines which promotes leukocyte recruitment in the intimal layer. These leukocytes, after the adhesion to the endothelium, penetrate the intimate tunic. CONCLUSIONS: Therefore inflammatory processes promote the onset and evolution of atheroma and the development of thrombotic complications.

A new morphological classification during follow-up in patients with celiac disease: a three-dimensional observation by scanning electron microscopy.

The structure and ultrastructure of the villi of small intestinal mucosa was examined in 237 duodenal or jejunal biopsies taken from children with active celiac disease and during gluten-free diet. All biopsies were processed for light and scanning electron microscopy. Conventional histology showed four different morphological aspects: total and subtotal villous atrophy in patients on unrestricted diet, partial villous atrophy and normal mucosa during gluten-free diet. Scanning electron microscopy demonstrated that in active celiac disease the severity of the intestinal lesions was related to individual vulnerability to gluten. Our results showed that during dietary treatment the process of mucosal healing was constant and strictly time-dependent. Furthermore, the ultrastructural examination has been relevant in evaluating the evolution of the villous regeneration. In this study a classification regarding the healing process of the small intestinal mucosa correlated with the time of start of dietary therapy is proposed.

A technique for exposure of the glycoproteic matrix (zona pellucida and mucus) for scanning electron microscopy.

The fine structure of the zona pellucida (ZP) covering the oocyte and of the mucus covering the surface of the intestinal villi was investigated by using a new method employing ruthenium red (RR), saponin, and osmium-thiocarbohydrazide impregnation. The glycoproteic matrices both appeared constituted by thin filaments (ranging from 22 to 50 nm in thickness) anastomosed to form a very fine network. RR prevented the dissolution and/or alteration of glycoproteins and polyanionic carbohydrates induced by acqueous fixatives. Saponin was a detergent of the soluble proteins. Osmium-thiocarbohydrazide preserved the glycoproteic matrix filaments from the mechanical stress induced by dehydration and critical point drying and reduced filaments packing and shrinkage. The technical improvement was demonstrated by the following results: 1) a regular arrangement of the filaments network; 2) a thickness of mucus filaments smaller than that obtained with other methods of preparation; 3) a homogeneous thickness of ZP filaments. This method allowed a very detailed study of the fine structural organization of the ZP and intestinal mucus. Therefore, this technique can be useful for a better evaluation of the morphodynamic of these and other glycoproteic matrices.

Ultrastructural features of bovine cumulus-corona cells surrounding oocytes, zygotes and early embryos.

Integrated transmission and scanning electron microscopic (TEM and SEM) techniques have provided the first detailed description of the ultrastructural features of the bovine cumulus-corona (CC) cells surrounding oocytes at the time of final maturation, zygotes and early cleaving embryos (2/4 to 6/8 blastomeres). TEM revealed the presence of rough endoplasmic reticulum and Golgi complexes in the cytoplasm of CC cells surrounding immature, mature and fertilized eggs, and also revealed an increasing amount of smooth endoplasmic reticulation membranes, lipid droplets and mitochondria with villiform and/or tubular cristae in the cytoplasm of CC cells during maturation and fertilization of the oocyte. In addition, a loss of cell-to-cell junctions between CC cells was evident. TEM also demonstrated that a few residual CC cells were still associated with early embryos and that these cells showed rather degenerative or apoptotic patterns, the latter pattern also observed on cells associated with fertilized eggs. SEM revealed that the complex of CC cells of immature oocytes was compact with narrow intercellular spaces, which progressively enlarged in size around mature oocytes. This phenomenon is mostly due to the production of abundant extracellular matrix. Immature CC cell complexes possessed characteristic long and filiform microvilli whereas the surface of CC cells surrounding mature oocytes showed numerous blebs and occasional large cytoplasmic protrusions as well as microvilli. Zygotes and early embryos were covered with a few polyhedral CC cells possessing scarce and short microvilli and a large amount of pleomorphic blebs. This study demonstrated a precocious luteinization occurring in bovine CC cells at ovulation until zygote segmentation, and this process was associated with a progressive apoptotic mechanism that ended in the complete denudation of the zona pellucida covering the early embryo. The presence of CC cells around the maturing oocyte and fertilized egg could have important functions related to the microenvironmental requirements of ovum maturation as well as facilitating activities related to fertilization.

Focal cell contacts detected by ruthenium red, triton X100 and saponin in the granulosa cells of mouse ovary.

Granulosa cells in growing follicles of mouse ovary, observed after treatment with ruthenium red (RR) as described by Luft (1971a, b), appeared to be covered by a continuous well-defined layer. On the contrary, treating granulosa cells with 1% Triton X100 (Vaccaro and Brody, 1981), followed by RR staining, resulted in the complete extraction of the plasma membrane coat (Triton does not affect the basement membrane and extracellular matrix proteoglycans). The use of 0.02% saponin together, with the RR stain, or 0.1% Triton X100 followed by RR staining, allows good visualization of follicular basement membrane and extracellular matrix proteoglycans without destroying cell morphology. Using this technique, we observed the extraction of the plasma membrane coat, but focal RR-stained condensations that were unaffected by saponin or 0.1% Triton X100 treatment were observed between plasma membranes of granulosa cells located around the periphery of large Graafian follicles. In some cases, RR condensations were located at the apex of plasmalemmal evaginations, in proximity to adjacent granulosa cells. Focal condensations of RR stain were never observed in secondary follicles. Present evidence suggests that focal cell contacts are mediated by transmembrane intercalated glycoproteins or proteoglycans and consequently play a role in cell adhesion. Their presence among granulosa cells of only very large Graafian follicles may be related to the maturation process of granulosa cells.

The extracellular matrix of healthy and atretic mouse ovarian follicles studied by lanthanum nitrate and polycation ruthenium red.

Changes occurring in the fine structure of Zona pellucida and follicular fluid in growing and atretic follicles in mice were examined by electron microscopy using lanthanum nitrate and ruthenium red. The results showed that follicular fluid has a dense globular aspect in growing follicles but becomes more dispersed and globular-fibrillar in atretic follicles. It is formed by rounded negatively charged particles (20 nm in diameter) that are closely packed in growing follicles, but spaced and united by filaments in atretic follicles. These changes may be related to a changed role of granulosa cells and also to an altered passage of nutritive substances from the plasma into the follicle. The Zona pellucida is intensely colorable only in the atretic follicles, where it is composed of a dense globular fibrous matrix, but not in growing follicles, where it reveals a weak reaction with ruthenium red. While in the Zona pellucida of growing follicles the globular negatively charged particles (20 nm in diameter) probably due to sialic acid moieties are rarely evident, these particles become highly numerous and connected by microfilamentous structures in atretic follicles. The altered electrical charge of polyanions may reflect a different chemical permeability of the Zona pellucida. Then the increase of negative charged particles in the zona may have a role in a process of sequestration of the oocyte during atreasia.

Microfilaments in granulosa cells of rabbit ovary: immunological and ultrastructural observations.

The present observations demonstrated that the granulosa cells of developing follicles in the rabbit ovary bind antimyosin-like antibodies (AMA). The same granulosa cells studied with the electron microscope showed bundles of microfilaments in the cortical areas of the cytoplasm and within microvilli and large cellular evaginations. The immunochemical and ultrastructural results suggest that the ovarian follicles are not passive organs, but, containing contractile granulosa cells, are capable of producing "in vivo" discrete movements.

Gastric re-resection in emergency.

BACKGROUND/AIMS: We reviewed a consecutive series of patients with acute complications following gastric resection, in order to evaluate the role of gastric re-resection as the operation of choice in emergency. METHODOLOGY: Records of 90 patients with acute complications following gastric resection observed from January 1991 to January 1996 were retrospectively analyzed. RESULTS: Hemorrhagic events occurred in the vast majority of cases (87), either as acute complications after a long time since surgery (78 cases) or as early postoperative complications (9). Among late acute complications, three cases were related to bleeding cancer of the gastric stump. Anastomotic obstructions presenting as acute complications occurred in 3 cases. Emergency surgery was indicated in three cases of acute obstruction, in 1 case of bleeding cancer of the gastric stump, in 9 (12%) out of the 75 remaining late acute hemorrhagic complications and in 1 (11%) out of 9 early hemorrhagic complications. Completion gastrectomy was chosen in the one case of bleeding cancer of the gastric stump with indication for emergency surgery. Gastric re-resection was performed in 11 cases: 9 for hemorrhagic complications and 2 for obstructive acute complications. In two cases, one for hemorrhage and one for occlusion, other surgical procedures were carried out. CONCLUSIONS: Gastric re-resection can represent the most suitable operation in acute complications following gastric resection.

[Variations in serum sialic acid in patients operated on for colonic cancer]. / Variazioni dell'acido sialico nel siero di pazienti operati per cancro del colon.

Various Authors believe that an increase in serum sialic acid might indicate the cancerisation of the membrane. Serum sialic acid was assayed pre- and post-operatively in patients with colonic carcinoma and in a control group. Sialic acid concentrations were significantly higher in the cancer group and the data obtained confirm the view that the increase has an effect on immune response in tumor patients.

[Value of serology and endoscopy and clinical aspects in idiopathic rectocolitis]. / Valore di sierologia, endoscopia e clinica nella rettocolite idiopatica.

The clinical course of ulcerative colitis is characterized by recurring acute attacks and interspersed with periods of remission. The present study correlates the clinical, endoscopic and laboratory results in the follow-up of 22 rectocolitis patients in order to identify any data that might serve as a warning of recurrences. It is claimed that laboratory examinations like titering of alpha 1-antitrypsin, antithrombin III collagenase, C3 and C4 offer useful support for endoscopy which is generally considered the only reliable examination in the diagnosis of idiopathic rectocolitis.

[Involvement of the polypeptide system and ACE in Crohn disease]. / L'interessamento del sistema polipeptidico e dell'ACE nella malattia di Crohn.

The polypeptidyc system was studied and serum concentrations of ACE (Angiotension Converting Enzyme) were assayed in patients with Crohn's disease in the active and stabilising phases. The study was undertaken on the assumption that as a granulomatous condition Crohn's disease involves the polypeptidyc system in much the same way as in sarcoidosis. The results confirm the hypothesis revealing a significant increase in ACE levels in patients with the active disease, whereas levels in the stabilized disease were lower than in the controls. It is suggested that, as in sarcoidosis, ACE levels are useful in the diagnosis, prognosis and follow-up of Crohn's disease, especially in relation to the use of steroid drugs.

[Clinical and therapeutic aspects of stab and gunshot wounds]. / Aspetti clinici e terapeutici delle ferite d'arma bianca e da fuoco.

In the last three years (1992-1995) 130 stab (114) and gunshot (16) wounds were observed at and admitted to the Emergency Surgical Department of Fatebenefratelli Hospital of Milan. We observed a high incidence of non-EEC patients (62%). Imaging devices (US and CT scan) and surgical minimally invasive procedures have reduced open surgery rate with a remarkable reduction in drawbacks and mortality.

[Laparoscopic and thoracoscopic reintervention after mini-invasive surgery in emergencies]. / Reinterventi laparo e toracoscopici dopo chirurgia mini-invasiva in urgenza.

One-hundred-eighty patients underwent emergency minimally invasive procedures (laparo and thoracoscopy) in the last two years. In twelve postoperative drawbacks another surgical procedure was performed. Laparoscopic or thoracoscopic exploration was undertaken in 8 cases while open surgery was done in 4. Complications and surgical overtiming rate was higher after endoscopic than open surgery. This approach allows us to correct very few and selected disease when a skilled surgical team is available especially during reoperations.

[Carcinoids in emergency surgery]. / I carcinoidi nell'urgenza chirurgica.

The authors report their experience of three cases of carcinoids of the gastroenteric tract which required emergency surgery. Two patients presented symptoms of acute appendicitis caused by appendicular carcinoid, whereas the third presented an occlusive syndrome due to ileal carcinoid.

Detecting microcalcifications in atherosclerotic plaques by a simple trichromic staining method for epoxy embedded carotid endarterectomies.

Atherosclerotic plaques have a high probability of undergoing rapid progression to stenosis, becoming responsible of acute coronary syndrome or stroke. Microcalcifications may act as enhancers of atherosclerotic plaque vulnerability. Considering that calcifications with a diameter smaller than 10 mm in paraffin embedded tissue are rather difficult to detect, our aim was to analyze microcalcifications on semithin sections from epoxy resin embedded samples of carotid endarterectomies using an original trichromic stain (methylene blue--azur B--basic fuchsine--alizarin red). We have compared samples stained either with our method, methylene blue-azur B alone or with Von Kossa staining, and methylene blue-azur B -basic fuchsine alone or with Von Kossa staining. Our method resulted to be simple and fast (ca. 2 min), it gives a sharp general contrast for all structures and allows to easy identify collagen and elastin. In addition, gray-green colour associated to intracellular lipid droplets evidences foam cells, which are particularly abundant in endarterectomies samples. Mast cells and their metachromatic granules are also well recognized. Calcifications over 0,5 mm are clearly recognizable. In conclusion, microcalcifications are clearly distinguished from the extracellular matrix in spite of their reduced dimensions. Methylene blue--azur B--basic fuchsine--alizarin red method is easy to use, reproducible, and is particularly suitable for the identification of microcalcifications in the morphological analysis of atherosclerotic plaques.