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.

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.

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.

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.

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.

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.

Endomicroscopy with Fluorescent CD105 Antibodies for "In Vivo" Imaging of Colorectal Cancer Angiogenesis.

The aim of this case report was to evaluate the feasibility of in vivo acquisition of microscopic images using fluorescent CD105 antibodies for molecular imaging in human colorectal cancer. After excluding the presence of tissue autofluorescence, the antibody solution was topically administered through a spray-catheter. The targeted area was analyzed by eCLE and images were recorded. The fractal dimension of tumor vessels and the vessel density were determined using ImageJ software. Immunohistochemistry was used as a gold standard. In vivo CLE analysis of CD105 expression enabled the study of tumor vascular network, revealing a chaotic structure.

Iron Oxide Nanoparticles Biodistribution in an Experimental Pig Model - A New Approach for Delivery and Imaging.

Iron oxide nanoparticles (IONs) are of great interest in medicine, with great potential for imaging diagnostics, as well as therapeutic. Biomedical applications of IONs have been suggested for magnetic resonance imaging (MRI), with two available contrast agents on the market. However, new developments in biocompatibility and biodistribution are necessary as many new physiochemical features of coatings have been proposed for a good safety profile. MATERIALS AND METHODS: Our study objective was to assess a different setting in terms of biodistribution of IONs coated with citric acid on an experimental pig model, based on EUS-guided portal vein (PV) injection. Four pigs were subjected to EUS procedures and portal vein injection of an IONs solution. All animals were kept under surveillance for the next 24 hours and euthanized. Necropsy was performed and their organs were harvested, visualized with a 3T MRI scanner and sent to pathological examination. RESULTS: All pigs had no change in their behavior and no signs of complications were encountered. There were no problems in identifying the pig's PV under EUS-guidance. The IONs solution was clearly visualized on ultrasound live imaging, during EUS-injection. MRI and histopathological data confirmed all the deposits using Prussian Blue staining. CONCLUSIONS: This paper comes forward as a first phase of assessing new future therapeutic options and their distribution within the main organs depending on their characteristics. In our opinion this new distribution option has a strong incentive to the research of therapeutic and imaging areas and is worthy of further appraisal.

Stem cells, colorectal cancer and cancer stem cell markers correlations.

: The idea of stem cells as being progenitors of cancer was initially controversial, but later supported by research in the field of leukemia and solid tumors. Afterwards, it was established that genetic abnormalities can affect the stem and progenitor cells, leading to uncontrolled replication and deregulated differentiation. These alterations will cause the changeover to cancerous stem cells (CSC) having two main characteristics: tumor initiation and maintenance. This review will focus on the colorectal cancer stem cell (CR-CSCs) theory which provides a better understanding of different tumor processes: initiation, aggressive growth, recurrence, treatment resistance and metastasis. A search in PubMed/Medline was performed using the following keywords: colorectal cancer stem cells (CR-CSCs), colorectal neoplasms stem cells, colorectal cancer stem cell (CR-CSCs) markers, etc. Electronic searches were supplemented by hand searching reference lists, abstracts and proceedings from meetings. Isolation of CR-CSCs can be achieved by targeting and selecting subpopulation of tumor cells based on expression of one or multiple cell surface markers associated with cancer self-renewal, markers as: CD133, CD166, CD44, CD24, beta1 integrin-CD29, Lgr5, EpCAM (ESA), ALDH-1, Msi-1, DCAMLK1 or EphB receptors. The identification and localization of CR-CSCs through different markers will hopefully lead to a better stratification of prognosis and treatment response, as well as the development of new effective strategies for cancer management.

Immunohistochemical expression of TGF beta (TGF-ß), TGF beta receptor 1 (TGFBR1), and Ki67 in intestinal variant of gastric adenocarcinomas.

Although in the last decades the incidence of gastric cancer declined, at present it is ranked worldwide on the fourth place between all human cancer pathology. Also, it has an aggressive behavior, the majority of patients being diagnosed in advanced stages. One of the key factors to control survival improvement of those patients is to clarify the molecular mechanisms involved in initiation, progression, invasion, and metastasis of gastric cancer. We thus investigated the immunoreactivity for TGF-ß, TGFBR1, and Ki67 of 25 specimens of intestinal gastric adenocarcinomas, and compared this with the correspondent reactivity for three specimens of diffuse gastric carcinomas; in the end, we tried to establish a statistical correlation with major clinicomorphological parameters. As a result, we noticed that the highest reactivity was present in the diffuse type compared with the intestinal variant, in which the TGF-ß reactivity progressively increased along the normal epithelium-intestinal metaplasia-dysplasia-carcinoma sequence. Also, we found for intestinal variant that TGF-ß immunoreactivity correlated significantly with tumor degree of differentiation and proliferative activity measured based on Ki67 immunoreactivity. In conclusion, TGF-ß is implicated in the progression of intestinal type of gastric adenocarcinomas and its immunoreactivity assessment for these targets has a prognostic value.

Morphometric findings in avascular necrosis of the femoral head.

Avascular necrosis of the femoral head is an illness with a controversial etiology, the trigger event being the suppression of blood flow to the femoral head. The disease affects mostly young adults within their third and fifth decade, the majority of the patients being men. The main risk factors are trauma, chronic alcohol consumption, smoking, corticotherapy. The main goal of our study is to describe the morphometric changes found in the bone tissue of patients diagnosed with avascular necrosis of the femoral head, with different risk factors, by comparing the area of bone trabeculae inside the area of necrosis with that from the adjacent viable tissue. The morphometric study used biological material from 16 patients with ages between 29 and 57 years, who underwent surgery for avascular necrosis of the femoral head. They were admitted in the Orthopedics Department at the Emergency County Hospital in Craiova between 2010 and 2011 and were split into four groups. Group I presented trauma as the main risk factor, Group II had corticotherapy as the defining risk factor, Group III presented chronic alcohol consumption and Group IV was represented by the patients who smoked and exhibited chronic alcohol consumption. There was not a significant statistical difference between the areas of bone trabeculae of the four groups when we compared viable bone tissue to the necrotized one. Knowing the risk factors of the avascular necrosis of the femoral head is critical to the management of the disease, because diagnosing it in an early stage is a necessity for obtaining a good result for conservative treatment.

Chronic bronchitis: a retrospective clinicopathologic study of 25 cases.

Chronic Obstructive Pulmonary Disease (COPD) is one of the leading causes of morbidity and mortality in the industrialized and in the developing countries. Chronic bronchitis (CB) is one the three COPD clinico-pathological entities that in 2009 were estimated to be diagnosed in 9.9 million Americans. It is characterized by inflammation of the "bronchial tree" that results in tissue swelling and excessive secretions of mucus into the bronchi, with progressive airflow limitation. Our study aims to reveal the main morphological aspects of CB in our casuistry and to evaluate their correlation with major clinico-epidemiological parameters. Thus, we performed a retrospective clinical and morphological study on a group of 17 smoker patients with symptoms of chronic bronchitis, eight non-smokers diagnosed with chronic bronchitis and five non-smokers and asymptomatic subjects. We observed that CB developed especially in men of 65-year-old or older, especially in smokers with a median FEV1% at around 71. Histopathologically, patients with symptoms of CB, regardless of smoking status, presented on bronchial biopsies with focal squamous metaplastic change, goblet cell hyperplasia and enlargement of the bronchial gland mass because of the inflammatory process, consisting predominantly of mononuclear cells in the bronchial wall. The statistical testing proved a significant correlation between the densities of different inflammatory cell classes (with the exception of mast cells in the bronchial epithelium) and FEV1% values on epithelium and submucosa regions in all investigated groups.

Breast invasive lobular carcinoma: a retrospective clinicopathologic study of 25 cases.

Invasive lobular carcinoma (ILC) is the second most common type of invasive breast cancer, having distinct prognostic and biologic implications. As an objective of the present work, we analyzed the clinicopathologic characteristics and prognostic factor of this invasive breast cancer variant. Clinical and morphological data of 25 cases of ILC collected during 2006-2011 were reviewed. Histopathologically, 11 cases were of classic type, and the others were non-classic with solid and histiocytoid subtypes being mostly encountered. Overall the non-classic ILC type was diagnosed in more aged patients (with a median age at onset of 59 years), with a predominance for a more advanced tumor degree differentiation (78.5% as grade 2 and 3), in advanced pTNM stages (50% in stage III and IV), with 50% lymph node involvement and with over 70% ER and Her2 reactivity. Statistically, we found that for the solid variant prevailed a PR+ and Her2- status while in histiocytoid subtype the PR- and Her2+ immunoprofile was most encountered. We conclude that non-classic ILC type represents a distinct entity of invasive breast carcinoma with a worsen prognostic than the conventional ILC type.

Collagen IV and MMP-9 expression in hypertrophic gingiva during orthodontic treatment.

INTRODUCTION: In the present study, we aimed to assess MMP-9 and type IV collagen in the gingival tissue in patients with gingival overgrowth (GO) after orthodontic treatment, with or without clinical signs of inflammation to appreciate the role of balance between those two markers in the onset of GO during tooth movement. MATERIALS: Gingival tissue was harvested from 45 patients divided in three groups: 15 patients with orthodontic appliance that developed GO on at least two teeth; 15 patients with chronic gingivitis with GO; 15 patients (control group) with healthy periodontal tissues, without clinical gingival changes, in whom the first premolar was planned for extraction for orthodontic purposes (canine distalization). METHODS: The tissue samples obtained by gingivectomy were fixed in 10% neutral formalin solution for 48 hours and then included in paraffin with the usual technique. The histologic examination was performed using classic Hematoxylin-Eosin technique. Double immunofluorescence was performed for anti-MMP-9 and anti-collagen IV antibodies. RESULTS: MMP-9/collagen IV double stainings showed an increase of fibrosis and inflammation in different degrees. CONCLUSIONS: The mechanical stress induced by the orthodontic devices, might be the key players in driving both the inflammation and the fibrotic reaction.

VEGFR1 and VEGFR2 immunohistochemical expression in oral squamous cell carcinoma: a morphometric study.

Vascular endothelial growth factor (VEGF) is considered one of the main molecules involved in tumor angiogenesis and is largely expressed in oral squamous cell carcinoma (OSCC). His signal is transmitted intracellulary by binding with class III tyrosine kinase receptors, known as VEGF receptor family (VEGFRs). Therefore, we designed this study for quantification of VEGFR1 and VEGFR2 immunohistochemical expression in the tumor cells of OSCC, and compare this expression with clinicopathologic parameters. For this purpose, 46 formalin-fixed, paraffin-embedded tissue blocks of OSCC were processed by immunohistochemistry. The immunohistochemical signal was assessed by estimating the area of the objects and the medium pixel intensity per object, as the integrated optical density (IOD). In our study, VEGFR1 staining intensity was significantly higher for tongue localization, while VEGFR2 was higher for the lip. Both markers were higher expressed in the center of the tumor compared to the tumor front. Moderate differentiated tumors exert higher expression levels for VEGFR1 but lower for VEGFR2. pT1 tumors had higher VEGFR1 levels, and when lymph node involvement was present, this was accompanied by elevated expression levels for VEGFR2 and lower levels for VEGFR1. These results point to an inverse profile of these receptors in OSCC, suggesting their involvement in a sequential manner in VEGF signaling regulation. In conclusion, our study revealed that VEGFR1 and VEGFR2 correlate with tumor localization, tumoral area (front vs. center of the tumor), histological differentiation degree, and lymph node involvement, while only VEGFR1 correlated with pT stage.

VEGF expression in human brain tissue after acute ischemic stroke.

Ischemic stroke is the third most common cause of death in humans, requiring further studies to elucidate its pathophysiological background. One potential mechanism to increase oxygen delivery to the affected tissue is induction of angiogenesis. The most potent proangiogenic factor is VEGF. For this reason, our study investigated immunohistochemically VEGF reactivity in different cellular brain compartments from 15 ischemic stroke patients, as well as from 2 age control cases. By enzymatic immunohistochemistry, we investigate VEGF expression in different brain cell compartments and then we quantified its signal intensity by assessing integrated optical densities (IOD). To establish the exact cellular brain topography of VEGF immunoreactivity we performed double fluorescent immunohistochemistry series (VEGF÷NeuN, GFAP, CD68, CD105). In control samples, VEGF reactivity was observed especially in neurons from the Brodmann cortical layers IV to VI and in protoplasmic astrocytes from the deeper layers of gray matter and in endothelial cells from normal blood vessels because of systemic hypoxia generated after death. In acute ischemic stroke samples, this reactivity was noticed in all brain cellular compartments but with different intensities. The most reactive compartment was the neurons, the intensity of VEGF reaction decreasing with the lesional age from the core infarct toward intact adjacent brain cortex. With a lower intensity, VEGF reaction was noticed in astrocytes compartments, especially in gemistocytic astrocytes adjacent to the liquefaction zone. We also noticed a weak reaction in activated non-phagocytic microglia from the periphery of liquefaction zones, and high VEGF-CD105 colocalization values at the level of microvessels that surround the infarcted brain area. In conclusion, this reactivity could suggest that VEGF might exhibit neuronal and glial protective effects and also a neoangiogenic property in acute ischemic stroke, facts that may have significant therapeutically impact on these patients.

The utility of CD44, CD117 and CD133 in identification of cancer stem cells (CSC) in oral squamous cell carcinomas (OSCC).

One of the theories regarding oral carcinogenesis is that the tumor growth is dependent on cancer stem cells (CSCs) that have the capacity of self-renewal and of giving rise to more differentiated tumor cells, like the stem cells do in normal tissues. The most used methods of CSCs isolation are based on their identification based on the expression of different cell surface markers. The markers qualified for this purpose have been described originally in studies involving hematopoietic or embryonic stem cells. Thus, we were interested to study the expression of the most used CSCs surface markers for formalin fixed paraffin embedded tissue samples from oral squamous cell carcinoma (OSCC). We investigated by immunohistochemistry thirty tissue samples of OSCCs with different degrees of differentiation and different oral locations. We were interested to establish the tissular localization pattern for cells expressing CD44, CD133 and CD117 in tumoral samples. The results indicated that with the exception of CD44, the other two surface markers were expressed only in tumoral stromal cells. When we looked at their origin (by double immunohistochemistry for cytokeratins AE1-AE3, vimentin and CD34) we concluded that they are of mesenchymal nature. Also, we proved that some of these cells also co-expressed CD44 but were negative for CK5÷6. Moreover, some of the stromal cells that were positive to CD133 and especially for CD117 also had reactivity to tryptase showing their mast cell nature. In conclusion, our study proved that CD44 has limited utility in identifying oral CSCs, while CD117 and CD133 expression appears to be limited more in identifying mesenchymal stem cells.

Mucinous adenocarcinoma of the colon - a histochemical study.

Colorectal carcinoma is a major cause of cancer associated with a high rate of morbidity and mortality in the western world. One of the pathologic features considered to be important for prognostic is mucin production. Many authors confirmed that colon carcinomas with high mucin content tend to re-occur locally and carry a poor prognosis. For histochemical evaluation of mucin content, we investigated 149 patients who underwent surgical resection of sporadic colon adenocarcinomas, all over a 5-year period. For histological classification we used the WHO recommendation (2000) and to be more accurate we sub-classified mucinous adenocarcinomas by morphometrical analysis in three categories: pure mucinous, with extracellular mucin more than 80% of the tumoral volume; mixed type, with 50-80% extracellular mucin; and mixed type with less than 50% extracellular mucin. For histochemical investigation, we used stains such as: mucicarmine, PAS ÷ Alcian Blue and High Iron Diamine ÷ Alcian Blue. Our study proved the predominance of mixed mucinous adenocarcinomas with less than 50% extracellular mucin, followed by the pure mucinous type. From the biochemical composition's point of view, the predominant cases were those with acidic mucins, especially in pure mucinous adenocarcinomas (>90%), while those with mixtures of acidic and neutral mucins were present in 62% of the cases. In addition, our study showed the prevalence of sialomucins over sulphomucins (68%), particularly in pure mucinous adenocarcinomas (77%). Clinical pure mucinous forms were detected mainly in advanced stages, but in terms of lymph node metastasis rate, they were secondary after mixed type with 50-80% extracellular mucin.

Fractal analysis differentiation of nuclear and vascular patterns in hepatocellular carcinomas and hepatic metastasis.

Hepatocellular carcinoma (HCC) currently represents the fifth most common cancer worldwide, while being the third leading cause of cancer death. Fractal analysis is a novel tool used in quantitative and qualitative image assessment. Vascular patterns and cellular nuclei particularities in tumoral pathology make ideal candidates for this technique. Our aim was to apply fractal analysis in quantifying nuclear chromatin patterns and vascular axels in order to identify differences between images of primary HCC, liver metastasis (LM) and surrounding normal liver tissue. Formalin-fixed, paraffin-embedded tissue sections from 40 cases of HCC and 40 LM of various origins were used. We performed Hematoxylin staining for nuclear chromatin as well as immunohistochemical staining for vascular patterns. High-resolution images were captured; nuclear and vascular morphologies were assessed on binarized skeleton masks using the fractal box counting method. Analysis was performed using the free, public domain Java-based image processing tool, ImageJ, which provided the fractal dimensions (FDs) for each studied element. Statistical analysis was performed using the ANOVA test with Bonferroni post-tests and t-tests for paired samples. Fractal analysis of vascular patterns clearly differentiated between tumoral tissue and normal surrounding tissue (p<0.01). Further analysis of nuclear FDs improved the specificity of these results, providing clear differentiation between pathological and normal tissue (p<0.01). When comparing primary HCC images with metastatic formations, we encountered statistically significant differences in nuclear chromatin assessment. However, blood vessels had a higher FD in primary tumors when compared with liver metastasis (p<0.05) and also allowed for a differentiation between primary liver tumors with and without neurodifferentiation. Fractal analysis represents a potent tool for discriminating between tumoral and non-tumoral tissue images. It provides accurate, quantifiable data, which can be easily correlated with the pathology at hand. Primary and metastatic liver tissue can be differentiated to some extent, however further studies, possibly including other variables (cellular matrix for instance) are needed in order to validate the method.