CD20cy and CD45RO immunoexpression in early rheumatoid arthritis synovium.

In this paper, we studied 15 cases of early rheumatoid arthritis presenting with inflammatory lesions in different degrees of evolution. We want to highlight B- and T-lymphocytes in synovial tissue collected from patients diagnosed with early rheumatoid arthritis, to establish the pattern of their distribution, possibly in relation to local neovascularisation to determine the role played by these types of cells. The pathological samples were represented by synovial membrane biopsy fragments, which were examined by histopathological and immunohistochemical methods. We noticed a perivascular distribution of lymphocyte infiltrate, up to formation of lymphoid follicles with germinal centers. There is a close interdependence between B- and T-lymphocytes in these lesions, and their presence in the synovial membrane in relation to newly formed blood vessels facilitates their action and their chemical mediators. Studying the interdependence of different types of lymphocytes and their connection with blood vessels may generate new therapeutic targets.

The utility of p16, E-cadherin and Ki67 in cervical squamous intraepithelial lesions diagnosis.

In this study, we included 26 cases diagnosed as squamous intraepithelial lesions, which were examined histopathologically, and in terms of p16, E-cadherin and Ki67 immunoexpression. In low-grade lesions, p16 expression was limited to one third below the epithelium, E-cadherin has a membranous pattern and Ki67 proliferation index had low values. In high-grade lesions, the p16 diffuse stain was present in two thirds or all epithelium layers, E-cadherin expression became aberrant, with membranous and cytoplasmic pattern and Ki67 proliferation index was high. These biomarkers have proven useful to accurately assess the extent of lesions and to identify lesions with high risk of progression.

E-cadherin and p63 immunoexpression in dysplastic lesions and urothelial carcinomas of the bladder.

In this study, we investigated 30 cases of dysplastic lesions and bladder carcinomas. The lesions presented different degree of differentiation and depth of invasion. The aim of this study was to appreciate the immunoexpression of some proteins, like E-cadherin and p63, which are implicated in the cellular adhesiveness and the normal development of the urothelium, depending on the degree of tumoral differentiation and the depth of invasion. The immunoreactivity of the markers was qualitative and quantitative evaluated. Both markers presented an immunoexpression diminishing with depth of invasion, the carcinomas with muscularis propria invasiveness having a minimum intensity reaction (0 or 1), and a medium percentage of marked cells by 10-50% for E-cadherin, and 30% for p63. The immunostaining intensity for E-cadherin decreased with the tumoral degree of differentiation, the poorly differentiated carcinomas having a reaction with minimum intensity (0 or 1). The p63-immunoexpression was correlated with the degree of differentiation in superficially cases of chorion invasiveness carcinoma. The immunostaining intensity in cases with muscularis propria invasiveness was diminished as the number of epithelial layers becomes bigger, but there was no variation depending on tumoral differentiation. In conclusion, E-cadherin and p63 are implicated in tumoral progression and may be used as a prognostic markers.

Immunohistochemical aspects of the evaluation of the inflammatory answer of the dental pulp.

The inflammatory answer of the dental pulp in front of an antigen presents a series of particularities caused by its special anatomical conditions. The evaluation of the inflammatory answer in the evaluation tests of the biocompatibility of the dental materials is made by histological methods without defining precise quantitative criteria to measure the pulp reaction. In this study, we followed the reaction of the pulp tissue to five antibodies (CD20, CD45Ro, CD4, CD8, and CD68) in order to evaluate the inflammatory aspects of the dental pulp. We did not find positive answers for the CD20 protein, specific for B-lymphocytes, for the fragments of normal pulp tissue taken into study. Even if they are in small number among the pulp cells, the T-lymphocytes that express the protein CD45Ro may be also found in the normal dental pulp. Of the two subsets of T-lymphocytes, we found positive answers on the studied preparations only for the CD8 protein. For the CD68 protein, strongly expressed by the macrophages, we obtained positive results both for the inflammated pulp tissue, and for the normal dental pulp, yet in a very small amount. The use of immunohistochemical techniques, with well-defined markers of the pulp inflammation, can offer better results for a highly accurate evaluation of the inflammatory answer of the dental pulp.

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.

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.

Immunohistological correlation in different stages of squamous cell carcinoma.

Squamous cell carcinoma is one of the most frequent cutaneous carcinomas, this neoplasic process inducing cellular and tumoral immune response modifications. Our study refers at 60 patients, squamous cell carcinoma diagnosed, at whom we determined IL-2, IL-6 and TNF-alpha, using ELISA technique. The discovered results were different, depending on the differentiation form. The cellular immune response presented important modifications only in poor differentiated form of the disease.

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.

The study of E-cadherine and CD44 immunoexpression in oral squamous cell carcinoma.

In spite of the progresses achieved in surgical treatment, radiotherapy and chemotherapy, the survival rate in oral squamous cell carcinoma (OSCC) remain unchanged among past three decades, which made this neoplasia a major problem of health in the entire world. There were investigated 42 cases of OSCC with different sites and various grades of differentiation, by histological and immunohistochemical techniques (by LSAB/HRP method), using E-cadherine and CD44. Considering the differentiation grade, the cases were histopathologically classified as 9 cases of well-differentiated squamous carcinoma, 14 cases of moderately differentiated squamous carcinoma, and 19 cases of poorly differentiated squamous carcinoma. The E-cadherine immunoexpression study indicated an immunostaining degree 3 in well-differentiated squamous carcinoma, 2 in moderately differentiated squamous carcinoma, and 1 in poorly differentiated squamous carcinoma. The results of CD44 immunostaining indicated in most of the cases an immunostaining degree 2, especially in moderately differentiated OSCC. The immunostaining degree 3 corresponded to nine cases of well-differentiated OSCC, and other two cases of moderately differentiated OSCC. Immunostaining degree 1 corresponded to poorly differentiated OSCC. The results indicate the possibility of using the two-immunohistochemical markers as prognostic factors in OSCC.

Immunohistochemical expression of vascular endothelial growth factor (VEGF) in intestinal type gastric carcinoma.

BACKGROUND: Gastric cancer still represents a difficult problem in the field of oncology, in terms of morbidity and mortality. The local progression and systemic spread is significantly influenced by tumor angiogenesis and lymphangiogenesis. In spite of many studies on the topic, data about the significance of growth factors in gastric cancer is controversial. AIM: to investigate the immunohistochemical expression of VEGF and to evaluate the relationships with the tumors stage and grade. MATERIAL AND METHODS: The immunohistochemical expression of VEGF was investigated on 80 patients with intestinal type gastric carcinoma. Specimens were fixed in buffer formalin, embedded in paraffin, and sections were stained with Hematoxylin-Eosin and immunohistochemistry was performed for VEGF (clone VG-1). Evaluation was performed using the VEGF score, based on the intensity of reaction and percent of positive cells. RESULTS: The reaction for VEGF was positive in 52 from 80 cases (70%). The final product of reaction was found in the cytoplasm of tumor cells, with granular pattern. Positive reaction was also found in eight from 28 cases with associated intestinal metaplasia, and in six from nine cases with gastric dysplasia. In the adjacent apparently normal mucosa, the reaction was positive in hyperplastic gastric pits and parietal cells. A strong correlation was found between VEGF expression and lymph node status and grade of the primary, but not with the stage of the tumor. CONCLUSIONS: The investigation of the immunohistochemical expression of VEGF in the intestinal type of gastric carcinoma showed positive reaction in 70% of the cases. It was demonstrated the expression of VEGF in intestinal metaplasia and gastric dysplasia, which could signify an early angiogenic switch during tumorigenesis.

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.

The endoscopic and morphological forms of early gastric cancer.

The early gastric cancer is an endoscopic notion in which gastric cancer is strictly placed to mucosis and submucosis without extensive manifestations. It is the form with favorable prognosis and better survival at 5 and 10 years. Our study tries to systematize the debut forms of early gastric cancer and their association with the lesions with malignisation risk. We also try to evaluate the incidence of endoscopic and histopathologic forms of early gastric cancer found in an internal medicine division. Our study included 435 patients with gastric cancer endoscopic and histologic diagnosed. Statistically, 64.36% were men and 35.64% were women, the mean age 48 +/- 7 years. The endoscopic forms of early gastric cancer were type I: protruded in 19 cases, type II: superficially in eight cases, type III: excavated in six cases. Early gastric cancer is diagnosed with difficulty, it represents in 7.58% of the gastric cancer, being most frequently asymptomatic. The endoscopic forms frequently found in early gastric cancer in the population were type I: protruded and type IIa: superficially elevated. The histopathological examination is compulsory at this form of gastric cancer, while in advanced gastric cancer endoscopy is often sufficient for diagnosis. Analysing the histopathological results of cases diagnosed with early gastric cancer we found: 22 cases with intestinal type and 11 cases diffuse type. Microscopically, 15 were intramucosal and 18 had submucosal invasion. I and IIa lesions were predominantly located at the antrum and are histologically differentiated adenocarcinoma. Differentiated carcinoma frequently produces an elevated lesion and the border is well demarcated. There are frequent opportunities to detect gastric cancer in the early phase and the patient can expect a complete cure by the surgical operation or endoscopical mucosal resection.

Diagnosis problems in a case of minimal deviation adenocarcinoma of the cervix.

We present the diagnostic problems in a case of minimal deviation adenocarcinoma of the cervix. Histopathologic exam of the tumor, made on serial sections, revealed a dense and profound proliferation of the glandular structures that were lined by endocervical type epithelia with minimal cellular and nuclear atypia. The aspect suggested the diagnosis of minimal deviation adenocarcinoma endocervical type; in order to confirm the diagnosis we immunohistochemical investigate the tumor for CEA, CA125, Ki67, ER and PR. The results indicated focal positivity for CEA, CA125 lack of immunostaining for ER and PR and Ki67 positivity with a low index of proliferation. We can conclude that all these markers are useful in the diagnosis, excluding the benign endocervical lesions.

P53 and PCNA immunoexpression in endometrial carcinomas.

This paper analyzes 18 endometrial carcinomas with different degrees of differentiation, the objective being to estimate involvement of p53 oncoprotein in the mechanism of endometrial carcinogenesis and the possible correlations with the tumoral proliferative activity evaluated by PCNA. The p53 immunoexpression was positive in 44.4% of the studied endometrial carcinomas, divided in three groups, the intensity of the immunostaining for p53 being increased for the low differentiated and undifferentiated endometrial carcinomas, whereas the differentiated endometrial carcinomas were moderate or low p53 positive. All the investigated tumors were PCNA positive, the PCNA index being of 40% in well-differentiated carcinomas, while in low differentiated and undifferentiated endometrial carcinomas the medium values of PCNA were of 60%, respectively 85%. Correlating the p53 and PCNA findings, we noted that PCNA was expressed especially in the cases with increased proliferative activity, without a significant statistic correlation between p53 and PCNA expression.

Gingival angiosarcoma: histopathologic and immunohistochemical study.

Angiosarcomas of the head and neck are rare tumors, the intra-oral localization being considered exceptionally in the specialty literature. The diagnosis of this kind of tumor in gingival location is difficult, both to the rarity of this lesion at this site, and to the various histopathological aspects. We present the case of 70 years old male patient clinically diagnosed with tumor of the alveolar crest of mandible, which, histopathologically, associated areas of predominant vascular proliferation, with areas of carcinoma and sarcoma. Immunohistochemical, the tumor was investigated for antibodies which prove the origin of proliferating cells: CD31, AE1-AE3 and vimentin.

HPV and p53 expression in dysplastic lesions and squamous carcinomas of the oral mucosa.

We studied 18 premalignant and malignant lesions of the oral mucosa, 3 dysplasias, and 15 oral squamous carcinomas with varied localizations: lips, palate, and tongue. Immunoassays for HPV were positive in 42.8% of the squamous carcinomas, without correlation with the pattern or the degree of differentiation of the lesions. P53 immuno-expression was positive in 2 of the 3 studied cases of dysplasia (66.6%) and in 13 from the 15 cases of squamous carcinomas (86.6%). The p53 immuno-positivity in both lesional categories, with a higher rate in carcinomas, indicates the fact that the p53 gene mutations can appear precocious in the oral carcinogenesis, but can not be used as malign potential prediction factor.

Salivary gland Basal cell adenomas--immunohistochemical evaluation of four cases and review of the literature.

OBJECTIVE: Evaluate the antigen profile of cellular population from basal cell adenomas. MATERIAL AND METHODS: Histopathological and immunohistochemical evaluation of four-salivary gland basal cell adenomas; the pathological samples were provided by the Oral Maxilla Facial Surgery Department of the Clinical County Hospital from Craiova. RESULTS: All basal cell adenoma exhibit differentiation toward 3 cell phenotypes: ductal luminal, basal and myoepithelial. CONCLUSIONS: The antigen profile of these tumors closely regard with other variants of salivary gland adenomas, such as pleomorphic adenoma and myoepithelioma. So, we can conclude that these tumors had a common origin, most probably from intercalated ducts and the proportion of those 3 cellular types, their cytoarchitectural arrangements and the quantity of extracellular matrix production can do the differentiation between them.

Cytokinin panel in rheumatoid arthritis and correlation with histological patterns of synovitis -- active type of disease.

UNLABELLED: Rheumatoid arthritis (RA) is a chronically inflammatory disease of the articular synovial, with severe, progressive and irreversible articular destruction. RA pathogeny implies an autoimmune mechanism, the role of cytokines resulting from the exaggerated production of some cytokines that act as pro-inflammatory agents, being directly responsible of certain symptoms and articular destructions, and/or inadequate inhibition of certain cytokines that act as anti-inflammatory agents. OBJECTIVE: We want to determine certain pro-inflammatory cytokines (tumoral-alpha -- TNF-alpha necrosis factor, interleukin-6 - IL-6, interleukin-8 - IL-8), anti-inflammatory cytokines (interleukin-10 - IL-10) and immunomodulators (interleukin-2 - IL-2) in some RA patients serum at the active stage of the disease and correlation with histological patterns of synovitis -- active type of disease. MATERIAL AND METHOD: The 37 patients have been grouped in stage I (9 patients), II (15 patients) and III (13 patients) according to the anatomical staging of the ARA (American Rheumatism Association), the serum levels of the cytokines being determined by ELISA technique. From the 37 patients clinical investigated only 12 were histopathological examined. RESULTS: The serum levels of IL-2 have been found low in patients with RA in the active stage of the disease, the lowest values having been determined at the patients in the 3rd stage of the disease, who also have the highest values of pro-inflammatory cytokines. In the case of IL-10 the lowest values have been found at patients in the advanced stages of the disease. In the serum of RA patients with follicular synovitis TNF-alpha was a dominant cytokine compared to patients with diffuse disease, but the greatest serum level was found in patient with granulomatous synovitis. Concentrations of IL-6 and IL-2 were highest in the serum of RA patients with follicular synovitis in comparison to patients with diffuse synovitis and could distinguish RA patients with these two histological variants of the disease. CONCLUSION: In the active stage of the disease the symptoms are a consequence of the interleukin pro and anti-inflammatory game: high serum levels of pro-inflammatory cytokines are accompanied by low serum levels of anti-inflammatory cytokines. Cytokines can be used as monitoring markers of the acutization period in RA, increase in serum levels of pro-inflammatory cytokines showing the progression from the inactive phase to a new period of activity of the disease. The association between distinct histological appearance of rheumatoid synovitis and serum cytokine profile and diverse clinical activity of disease seems to confirm its heterogeneity.

Tumoral stroma of salivary pleomorphic adenoma -- histopathological, histochemical and immunohistochemical study.

The aims of our paper were to establish the main histopathological, histochemical and immunohistochemical aspects of tumoral stroma from salivary pleomorphic adenomas. For this purpose we investigated 103 cases by the classical histopathological technique with paraffin embedding and staining with Hematoxylin-Eosin (HE), Hematoxylin-Eosin-Safranin (HES), trichromic Masson, trichromic Goldner Szeckelly, orcein and Periodic Acid Schiff-Blue Alcian (PAS-AA). Immunohistochemically, they were investigated for AE1-AE3, MNF116, CK8, EMA, vimentin, alpha-actin calponin, S-100, GFAP, collagen IV, and PCNA. The results of our study suggest the key role of neoplastic myoepithelial cell in the achievement of diverse morphological aspects of stroma in such neoplasms.