Lymphatic and blood microvasculature organisation in pulmonary sarcoid granulomas.

Pulmonary sarcoid granulomas are characterised by their elective distribution along collecting lymphatics. However, relationships between granulomas and intralobular lymphatics or blood microvascularisation have not been investigated. Therefore, we undertook a specific analysis of blood capillaries and lymphatics supplying sarcoid granulomas to identify additional clues to understanding the pathophysiogenesis of these lesions. Six pulmonary samples were immunolabelled with D2-40, anti-CD34 and anti-CD31 antibodies, paying particular attention to the relationships between lymphatics and granulomas, and the pattern of blood microvessels supplying sarcoid lesions. A morphometric study of granulomas included their distance to lymphatics and a three-dimensional reconstruction of a granuloma in its lymphatic context. Intralobular granulomas were closely associated with lymphatics; apart from a few granulomas, blood capillaries stopped at the outer border of the fibrous ring surrounding granulomas, and perigranuloma capillaries were particularly scarce. Our observations of the lymphatic and blood microvascular environment of intralobular pulmonary sarcoid granulomas provide evidence for the critical role of lymphatics in the emergence of these lesions. Moreover, pulmonary sarcoid lesions could be considered avascular structures, thereby providing new insights into the understanding of the granuloma physiology and the distribution of blood-borne therapeutic agents.

Pancreatic metaplasia of gastric mucosa associated with gastroduodenal ulcer.

Metaplasia represents the process of transforming a well-differentiated adult tissue into another type of adult tissue. Pancreatic metaplasia of the gastric mucosa represents the process in which the normal mucosa of the stomach is replaced with pancreatic formations, which mimic the structure of pancreatic acini. We describe the case of a male patient aged 39 who was admitted for abdominal pain, vomiting, hematemesis, melena, pale teguments, intense perspiration and nausea. The patient underwent surgery for suturing a perforated duodenal ulcer five years prior to this episode (2002). A gastric ulcer complicated with superior digestive bleeding and a chronic duodenal ulcer complicated with partial stenosis and perivisceritis were found during surgery. Gastric wall fragments were harvested and underwent usual histological techniques and immunohistochemistry. We found an ulceration from the gastric mucosa to the submucosa, covered by fibrino-leukocytic detritus. In the mucosal chorion we found numerous round or oval shaped nested formations which occupied the lower two thirds of the chorion, to the muscularis mucosae. Some metaplasic acini contain cells variable in shape, color and immunophenotype. Surrounding the nested acini we found tubular formations, formed of cubic shaped cells, representing excretory canals which were continued by gastric glands or opened directly in the crypts of the gastric epithelium.

Study of vascular microdensity in areas of cerebral ischemia on experimental model.

Stroke is the third leading cause of death and the first cause of disability with an increasing incidence, especially of the ischemic type. There is no effective curative treatment for stroke and therefore the therapy for this disease currently relies on identifying patients at risk and instituting preventive measures. Since ischemic stroke in middle cerebral artery territory (and with preferential localization in the striated nuclei) is the main type of stroke in humans, animal models obtained by surgical ligation of the middle cerebral artery (MCAO) are valuable tools for the fundamental study of this disease. In this study, we investigated the morphological and immunohistochemical remodeling of the tissue after stroke and in particular the vascular component in a MCAO murine model. The analysis included the sequential sacrificing of animals at 1, 14, 28 and 60 days post-stroke and brain processing for paraffin embedding and sectioning. Our results show a gradual revascularization of the lesion as we move away from the time of the surgical intervention. This effect is accompanied by the development of an increasingly dense glial scar at the periphery of the lesion. The perilesional area itself, the penumbra, is characterized by minimal histological changes (such as eosinophilic neurons), but also by an increased expression of activated caspase 3 as a sign of apoptotic neurons and glial cells. Our study confirms the potential of the organism in its attempt to revascularize the injured area, and raises questions on the role of the glial scar in limiting the process of neovascularization.

Histopathological changes in acute ischemic stroke.

We study here the histopathological changes in twenty-two cases of acute ischemic stroke. The average age of the patients was 62-year-old, and the interval from the onset of the disease to the death varied from 6 hours to 15 years. The brain lesions after acute stroke were observed in all regions. Their evolution allowed us to classify them in fourth stages. Phase one changes (1-2 days after onset) (n=2 patients) included red hypoxic and "ghost" neurons and other acute neuronal injury and spongiosis. The second phase (n=14 patients) was subdivided into: (a) a phase of acute inflammation (3-37 days after onset) (n=5 patients), where we observed especially features of acute inflammation together with coagulative necrosis, and (b) phase of chronic inflammation (10 days-53 years after the onset) (n=9 patients), in which prevail mononuclear and macrophage infiltrate, astrogliosis, spongiosis and neo-vascularization. In the third phase (26 days-23 years after the onset), we included six cases characterized by the absence of an inflammatory reaction, presence of cavitation, astrogliosis and macrophages. Our study describes the heterogeneity of brain injury after acute ischemic stroke with the participation of all brain components, and the chronology in which these lesions develop and evolve.

Fractal analysis of astrocytes in stroke and dementia.

The varied morphological forms in which astrocytes occur in brain of ischemic/hemorrhagic stroke and Alzheimer's disease (AD) patients are complex and the mechanisms that drive their formation are not yet properly understood. Subjective differences can be described between these pathologies in what it concerns astrocyte implication, but these have not been yet subjected to a morphometrical quantification. Here we apply a fractal dimension (FD) analysis algorithm to differentiate both between fibrous, protoplasmatic and activated astroglia; but also between the three pathological conditions studied. Analyzing more than 1000 astroglia, we show here first that FD can clearly differentiate between the three morphological subtypes. Second, we describe resemblances of the FD values for ischemic and hemorrhagic lesions, and significant differences when these are compared to AD patients. These results are further discussed and integrated in what it regards the preferential regions proved to be affected in these conditions, and which parallels our results. This work illustrates that fractal dimension analysis of astroglia is a useful method for quantitatively describing gliosis in different pathologies, and that it may offer more insight into the pathogenesis of brain diseases.

VEGF and VEGFRs expression in oral squamous cell carcinoma.

Oral cancer is an important cause of worldwide morbidity and mortality, with substantial economic, physiological, and psychosocial impacts due to its treatment modality and a great risk for recurrences and second primary oral squamous cell carcinomas (OSCC) development. Therefore, it is very important to understand the underlying cell biology of such tumors. It is now a well-accepted fact that angiogenesis is essential for the growth and metastasis of solid tumors, including head and neck squamous cell carcinoma. The main factor responsible for angiogenesis is VEGF and its receptors. It has been demonstrated that VEGFRs are also present on tumor cells themselves and other cells from the tumor microenvironment, in addition to tumoral endothelial cells (ECs). Therefore between these cells take place numerous and different interactions mediated via paracrine/autocrine pathways that promote angiogenesis, uncontrolled tumor proliferation and metastasation. In consequence, estimation of VEGF expression and its receptors became a reliable prognostic tool in OSCCS, predicting the poor disease-free survival, poor overall survival, and metastatic disease. Understanding the distribution and role of VEGF and its receptors in the progression of OSCC will be essential to the development and design of new therapeutic strategies.

Cathepsin-D expression in breast lesion: an immunohistochemical study.

Cathepsin-D (CathD) is an aspartyl lysosomal protease expressed in all tissues that might play a role in antigen processing, cell proliferation and tissue renewal, and activation of different pro hormones. The aim of our study was to compare the expression of CathD in most common breast tumors and tumor-like breast lesions. The study includes 21 patients with histologically verified breast lesions (adenosis, ductal hyperplasia, fibroadenomas, and different types of invasive carcinoma). We investigated the cathepsin-D expression in these breast lesions using immunohistochemistry (IH; paraffin-embedded tissues). Cathepsin-D staining within each lesion was assessed by estimating the area of the objects and the medium pixel intensity per object, as the integrated optical density (IOD). The immunostaining was more obvious in breast invasive carcinomas and macrophages. The reaction in tumor tissue was heterogeneous with little variation of staining intensity in positive tumor cells. Adenosis had the maximum area/signal intensity from all studied breast benign lesions (p<0.001, Student t-test). The general tendency (all benign lesions, lobular carcinomas and G3 ductal invasive carcinoma) was a more prominent representation of the cellular compartment. In the G3 ductal invasive carcinoma-type, the group of patients with metastases had a stronger expression in the cellular compartment. These results suggest that CathD expression was strongest in malignant than in benign breast disease, the positivity being present in both epithelial neoplastic and stromal cells. We also conclude that our procedure in IOD measurement is prone to less subjective-related biases, and thus more accurate and constant than other methods employed by other authors.

Histopathological and Morphometric Study of Fibrosis and Nuclear Pleomorphism in Dilated Cardiomyopathy.

Histopathological changes associated with dilated cardiomyopathy (CMD) are frequently nonspecific and often only present in the terminal stage of the disease. The study followed the histopathological and morphometric quantification of fibrosis and nuclear pleomorphism in CMD. We analyzed left ventricle myocardial fragments harvested during autopsy, from 35 cases with clinical diagnosis of CMD and 5 cases of normal myocardium. Fibrosis was present in all CMD cases, with higher values compared with control cases. Nuclear pleomorphism was identified in 18 cases (45%), two of the analyzed parameters, respectively the ratio of nuclear diameters and roundness of nucleus, revealing significant differences in CMD compared to the control cases. Myocardial fibrosis present in all cases of CMD represents a major feature of the disease. The nuclear pleomorphism due to the nuclei change in diameters and size was more pronounced in the vicinity of fibrosis areas, possibly related to this alteration.

Distribution of Aquaporins 1 and 4 in the Central Nervous System.

The aquaporins (AQP), a protein family, were first discovered in the early 1990s. The primary role of aquaporins is to facilitate water transport across multiple cell types. In the spinal cord and brain responsible for most of the water diffusion are AQP4 and AQP1. In this paper, we describe the structure, localization and role of this water channel family, especially AQP4 and AQP1. AQP4 is involved in various pathologies such as: stroke, brain tumors, Neuromyelitis optica (NMO), Alzheimer's Disease (AD), traumatic brain injury, Parkinson's Disease, hydrocephalus, schizophrenia, epilepsy, major depressive disorder, autism. Brain edema is the most important acute complication of the hypoxic-ischemic and it has no pathogenic treatment. Imaging and histopathology studies have shown that inhibition of AQP4 reduces brain edema.

Giant Rhinosinusal Inverted Papilloma.

Sinonasal papilloma is a benign tumor, derived from Schneiderian sinonasal epithelium. There have been described three histological subtypes: inverted, oncocytic and exophytic. The case presented here is A 66-year-old male patient, which was hospitalized in our Otolaryngology Department for a giant tumor, that was exteriorized from the left nostril, repeated epistaxis, nasal obstruction and anosmia. The computed tomography scan revealed an iodophilic and non-homogeneous tumor, with areas of necrosis, which included the entire left nasal cavity, with extension to the rhinopharynx and the left maxillary sinus. We completely removed the tumor by an endoscopic medial maxillectomy, with the subsequent histopathological examination revealing an inverted papilloma, with areas of low grade dysplasia and also areas with oncocytic Schneiderian papilloma. At the six-months postoperative control, there was no tumor recurrence. Major issues of this type of tumor is fast invasion capacity and numerous local recurrence. More recent studies have shown that these relapses are often overdue tumors.

Correlations Between Clinicopathological Features and the Vegetative Nervous System in Gastric Cancer.

Gastric cancer currently represents one of the most important public health problems. Recent studies have demonstrated the existence of strong correlations between the vegetative nervous system and the role it plays in the initiation of the oncogenetic process and the progression of cancer. The purpose of this paper is to demonstrate the involvement of the sympathetic and parasympathetic vegetative nervous system in the evolution of gastric cancer, according to the stage of tumor differentiation. In this current paper we have included a number of four patients diagnosed with gastric cancer post UGI (Upper Gastrointestinal Endoscopy) and have analyzed relations that exist between the tumor differentiation degree and the metanephrine and normetanephrine serum level in the blood of the patients. Following the research, we have observed an increased value of the metanephrine and normetanephrine serum level in the patient which displayed the lowest degree of differentiation.

Immunohistochemical characterization of tumoral vessels in oral squamous cell carcinoma.

Tumors require a blood supply for growth and hematogenous dissemination. Angiogenesis is one of the mechanism by which tumors acquire their microcirculation. Structurally and functionally, these newborn vessels are abnormal, showing increased permeability, delayed maturation, and potential for rapid proliferation. Such vascular defects could be an explanation for the aggressivity of oral squamous cell carcinoma (OSCC). For these reason we studied the morphology of tumoral vessels in such tumors by using immunohistochemistry and immunofluorescence. Forty formalin-fixed, paraffin-embedded tissue blocks of OSCC were processed for double enzymatic and fluorescence immunohistochemistry. We were interested in analyzing the tumor vessel architecture, and their maturity and activity in such tumors. The tumor vessel architecture had a chaotic pattern, mostly of different sizes, aberrant morphology, tortuous, without clear lumen, and irregularly branches. Regarding pericytes recruitment, the immature and intermediate vessel types (both negative to smooth muscle actin-SMA) were the most numerous type of tumoral vessels. The mature ones (positive to SMA) were readily more numerous at the invasive front of OSCC (85.4 vessels/4 mm(2) +/- 38.3), especially in poor differentiated tumoral type. Investigation of the tumor vessel basal membrane, as reactivity for collagen IV, revealed variability in thickness (2.59 microm +/- 0.48), small surface projections, discontinuities and loose associations with endothelial cells; these abnormalities being more obviously at the tumor-host interface and in poor differentiated OSCC. The most active angiogenesis was noticed in poor differentiated OSCC (0.23 +/- 0.04), at the tumor-host interface with the immature and intermediated vessel as the most active tumor vessel types. In conclusion, our study revealed some peculiar structurally and functionally defects of tumor vessels in OSCC, changes that could be selective targets for the new developing antiangiogenic drugs.

Annexin A3 expression after stroke in the aged rat brain.

In an effort to identify new proteins involved in functional recovery after cerebral ischemia, young (3 months) and aged (18 months) male rats were subjected to middle cerebral artery (MCA) occlusion. Brains were harvested at 3- and 14-days post ischemia and proteins from the peri-infarcted and the corresponding contralateral area and total proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis followed by mass spectrometry analysis. Annexin A3 (ANXA3) was identified as one upregulated protein in the post-ischemic rat brain. Using western blotting, real-time PCR and immunohistochemistry, we confirmed that at 3-14 days post-stroke, ANXA3 expression in the peri-infarct area was consistently increased over the corresponding area of control rats. Double staining revealed that ANXA3 is produced by activated microglial cells. We found that aged rats also had more newly proliferating cells expressing ANXA3 than young rats do. Occasionally, ANXA3-immunopositive cells wraped around neurons, suggesting that annexin A3 may be involved in the removal of dying neurons after stroke.

Endoglin (CD105) and microvessel density in oral squamous cell carcinoma.

Recent studies revealed that CD105 is intensively expressed in tumor vasculature, and may be an important prognostic indicator for the outcome in a number of malignancies. The aim of our study was to evaluate the CD105 expression and microvessel density in oral squamous cell carcinoma (OSCC). Nineteen surgical specimens with OSCC were immunohistochemical analyzed with CD105 (Endoglin). We determined the microvessel density (MVD) by "hot spot method". Endoglin was intensively expressed in vessels from the inner and invading front of all investigated OSCC. The highest value of MVD, about 30.89 +/- 22.4, were record in peritumoral area of OSCC, since intratumoral MVD average was about 10.18 +/- 4.7. We did not observe any significant association of MVD with age, sex, primary tumor's location, clinical stage or differentiation grade. In conclusion, CD105 expression is up-regulated in OSCC, and has a significant role in the development of such malignancies.

Chronic tonsillitis: histological and immunohistochemical aspects.

OBJECTIVES: Chronic tonsillitis represents the most common inflammatory lesions of the pharynx determining numerous local or distant evolutive complications. We decided to study the histological and especially immunohistochemical expression of this pathology. MATERIAL AND METHODS: We have studied 112 surgical samples representing tonsils resected from 56 patients with chronic tonsillitis. The tonsillectomies were performed in the ENT Clinic of the Emergency County Hospital of Craiova, between 01.01.2007--31.12.2007. The processed histological samples were stained using Hematoxylin-Eosin, light green trichromic and argental impregnation. For the immunohistochemical study, we used LSAB method with CD20, CD45 RO, CD68 antibodies in order to reveal and differentiate T- and B-lymphocytes and also the macrophages. RESULTS: In all samples, we found hyperplasia and hypertrophy of the lymphoid follicle with excessive developing of the clear germinal center as a normal reaction to antigens presence. In some cases, we remarked micro hemorrhages and hematic extravasations inside the follicles, probably due to the excessive virulence of the pathogens causing endothelial lesions. The conjunctive stroma was enriched in collagen fibbers, in some cases organized in strong fascicles with an obvious tendency to divide the tonsils in lobules. The young fibroblastic type cells were numerous. The specific reticulin fibbers had a low representation being disorganized. The immunohistochemical study proved that the clear center of the lymphoid follicles was occupied by B-lymphocytes, but the T-lymphocytes were present in the cortical region of the follicles, perifollicles and in the surface epithelium. CONCLUSIONS: In some pathological cases, the predominant cellular population of the clear center was formed by T-lymphocytes.

Histopathological Aspects of the Myocardium in Dilated Cardiomyopathy.

Dilated cardiomyopathy is the most common form of cardiac muscle disease, accounting for approximately 60% of all cardiomyopathies. We proposed to identify histopathological changes of the myocardium in dilative cardiomyopathy. This study comprised a total of 19 cases, represented by myocardial fragments from deceased patients with diagnosis of dilated cardiomyopathy. Histopathological analysis allowed changes to be observed for both myocytes and myocardial interstitial components. We have found a combination of hypertrophic, atrophic and normal myocardocytes, or associated with the presence of hydropic changes. We rarely identified the aspect of myocytosis, cytoplasmic accumulation of lipofuscin pigment or mucinous material, and variable nuclear pleomorphism. At the interstitial level we noticed changes in fibrosis, lipomatosis and rarely the presence of inflammatory infiltrate. Histopathological characteristics of the myocardium in dilated cardiomyopathy are numerous but nonspecific, similar to those in the terminal stages of other cardiac diseases.

Diagnostic of Early Onset Polycystic Kidney Disease in Neonates.

Polycystic kidney disease represented by autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD) have a major impact of mortality in children. We conducted a study of a premature infant with an estimated gestation date of 32 weeks with a presumptive prenatal diagnosis of right polycystic kidney. A 28-year-old primigravida with pre-eclampsia was admitted at the gynecology unit of Clinical Emergency County Hospital of Craiova. The clinical examination revealed a large abdominal distention due probably to the right polycystic kidney, suspected on prenatal ultrasound and radiography. The preterm neonate undergone right nephrectomy 5 days after birth. Histopathology of the kidney was performed in the Pathology Department of the Emergency County Hospital of Craiova and in the Center for Microscopic Morphology and Immunology of U.M.F. of Craiova. Microscopy revealed dilated cysts lined by simple cuboidal or flattened epithelium, and islets of remnant kidney parenchyma separated by edematous stroma. Immunohistochemistry for CD34 revealed incomplete blood arcades which did not seem to be in contact with all the tubular elements of the parenchyma, when compared to a control age-matched kidney. The patient had a favorable postoperative evolution, she was clinically stable on discharge from the hospital with a follow-up strategy including genetic testing.

Modified variant of the Rieske iron-sulfur protein in the hippocampus of kindled rats and human epileptic patients.

Kindled seizures are widely used to model epileptogenesis, but the molecular mechanisms underlying the attainment of kindling status are largely unknown. Recently we showed that achievement of kindling status in the Sprague-Dawley rat is associated with a critical developmental interval of 25 +/- 1 days; the identification of this long, well-defined developmental interval for inducing kindling status makes possible a dissection of the cellular and genetic events underlying this phenomenon and its relationship to normal and pathological brain function. Now we report the identification, by proteomics, of a modified variant of the Rieske iron-sulfur protein, a component of the mitochondrial cytochrome bc1 complex, whose isoelectric point is shifted toward more alkaline values in the hippocampus of kindled rats. By immunohistochemistry the Rieske protein is well-expressed in the hippocampus except in the CA1 subfield, a region of selective vulnerability to seizures in humans and animal models. We also noted an asymmetric, selective expression of the Rieske protein in the subgranular neurons of the dorsal dentate gyrus, a region implicated in neurogenesis. Abnormal changes in Rieske protein immunoreactivity also were found in sections obtained from human epileptic patients. These results suggest that the Rieske protein may play a role in the response of neurons to seizure activity and could give important new insights into the molecular pathogenesis of epilepsy.

The immunohistochemical profile of the adenocarcinoma of upper gastric pole.

Although gastric adenocarcinoma continue to be the second continues to be the second cause of death worldwide, its incidence and mortality appear to have decreased in recent decades. Despite this decline, adenocarcinomas from proximal stomach tend to be more frequent during the last three decade. Adenocarcinomas with this location it seems that are a different, specific subtype of gastric carcinoma. The purpose of this study was to clarify the differences between gastric adenocarcinomas from upper and distal gastric pole using the immunohistochemistry. For this reason, we investigate histopathological and immunohistochemically 77 cases of upper gastric pole adenocarcinoma selected from a number of 472 gastric tumors. The immunohistochemistry was performing only in 32 cases by ABC technique with the following primary antibodies: Cytokeratin 7, Cytokeratin 19, Epithelial Membrane Antigen (EMA), Carcinoembryonic Antigen (CEA), Lysozyme, Vimentin, p53 protein, CD34 and Ki67 antigen. The acquired results do not distinguish a peculiar immunohistochemically profile unlike distal gastric adenocarcinomas. Nevertheless, we pointed out the predominance of diffuse adenocarcinomas type according to Laurens classification, which immunohistochemically were strong positive to cytokeratins, EMA, CEA and lysozyme. Moreover, investigation of some antigens likes lysozyme, p53, Ki67 and CD34 seems to be useful for prognostic estimation of carcinoma with this topography.

In Vivo Experimental Study of an Endoscopic Ultrasound Multifunctional Radiofrequency Ablation Probe.

The aim of our study was to test the feasibility of a new developed RFA probe made especially for EUS use and also capable of injecting iron oxide nanoparticles within the targeted liver area. The procedures were performed on domestic pigs, divided in groups: A.liver RFA was performed; B -IONs were injected in the liver followed by EUS-RFA in the same area; C.local EUS-guided liver IONs injection were performed. After EUS measurements for the ablation areas, group A had a mean of 4.9 cm, while group B had a mean of 5.2 cm (Fig.3, 4). IONs exposure was on a median area of 3.1 cm. EUS imaging pointed out a regular oval shape in group A, and a slightly irregular outline on group B, with more echo bubbles around. MRI sections revealed different patterns for each group separately. In group A and B, RFA lesions were easily identified with specific liver parenchyma changes. Group B revealed few deposits of nanoparticles further away from the targeted point. The last group pointed out a large amount of IONs within the injection region and a larger amount of dispersed IONs within the liver than group B.