Ki-67, p53 and bcl-2 analysis in colonic versus rectal adenocarcinoma.

This paper develops a comparative study between similar subtypes of colonic and rectal adenocarcinoma, based on their immunohistochemical profiles for Ki-67, p53 and bcl-2 markers, in order to evaluate the prediction value for the investigated markers, according to the histologic subtype and location. Thirty cases of adenocarcinoma were investigated, 15 with colonic and 15 with rectal location. For both locations, the cases included five well-differentiated, five moderately differentiated and five low differentiated subtypes. The immunohistochemical investigation was performed using Ki-67, p53 and bcl-2 antibodies (DAKO) and streptavidin-biotin method (LSAB Kit, DAKO). The semiquantitative analysis of the immunohistochemical reactions was based on the Ki-67 and p53 index, respectively, counted as number of positive cells from 100 positive and negative cells. For bcl-2, the reaction was considered positive, respectively negative, for a percentage of positive cells higher, respectively smaller, than 5%. All 30 cases (100%) were positive for Ki-67. The mean value of Ki-67 index for colonic, respectively rectal adenocarcinoma was 55.8%, respectively 59.6%. No statistical correlation was found between the proliferative activity and location (p=0.502). Ten cases (66%) of colonic adenocarcinoma were positive for p53, with a mean value of 42.5% for p53 index. Eight cases (53%) of rectal adenocarcinoma were positive for p53, with a mean value of 21.1% for p53 index. There was a statistic correlation between the apoptotic activity and location (p=0.005). The positive reaction for bcl-2 was present in seven (46.6%) and nine (60%) from the 15 cases of colonic and, respectively, rectal adenocarcinoma. The statistic analysis revealed that bcl-2 cannot be significantly associated with any of the two locations (p=0.48144, 95% CI). The use of the Wald tests permitted the assessment of the predictive power for the investigated markers according to the pathologic subtype and location. Thus, p53 is on the first position (W=16.56, p=0.00004, 95% CI), followed by Ki-67 (W=4.49, p=0.034, 95% CI), whereas bcl-2 cannot be considered a predictive factor (W=2.5, p=0.107, 95% CI). The immunohistochemical evaluation of Ki-67, p53 and bcl-2 yields refined information on colorectal tumor biology. Our study confirms, from the statistic point of view, the role of p53, followed by Ki-67, as predictive factors.

T-cell primary cutaneous lymphomas. A clinicopathological and immunohistochemical study.

Within the large framework of the lymphoproliferative diseases, the primary cutaneous lymphomas are distinct pathologic conditions, defined by particular morphologic, immunologic, genetic, and clinic criteria. The study aimed to create the first clinicopathological and immunohistochemical profile of primary cutaneous lymphoma for a Romanian region. We investigated a series of 16 cases (diagnosed during a 5-year period) in accordance with the general principles of primary cutaneous lymphoma management. The methods included the clinic and morphologic exams, the latter relying on standard and immunohistochemical staining. The results revealed that all studied cases were T-type lymphomas, in terms of the WHO-EORTC classification. Most of these cases were diagnosed as mycosis fungoides; the group also included cases of Sezary syndrome, as well as rare entities such as: mycosis fungoides associated with follicular mucinosis and subcutaneous panniculitis-like T-cell lymphoma. Our discussions focused on the role of the clinicopathological assessment for the primary cutaneous lymphoma diagnosis and emphasized the importance of the immunohistochemical investigation. Compared with the previous Romanian researches on this topic, presenting only isolated cases, the current study develops a new level of analysis, based on the rigorous monitoring of a relatively large geographical area, for a long time horizon.

Hyperplasia, metaplasia, dysplasia and neoplasia lesions in chronic cholecystitis - a morphologic study.

The aim of the study was to analyze the association between chronic cholecystitis, premalignant lesions and gallbladder cancer. The group consisted in 3901 cases of cholecystectomies, diagnosed as acute cholecystitis (250 cases - 6.4%), chronic cholecystitis (3619 cases - 92.8%) and gallbladder carcinoma (32 cases - 0.8%). Chronic cholecystitis associated premalignant lesions as follows: hyperplasia in 124 cases (7.8%), metaplasia in 86 cases (5%) and dysplasia in 10 cases (0.4%). Only in nine cases, the diagnosis of gallbladder carcinoma was formulated presumptively, before surgery; for the other 23 cases this diagnosis was established after the pathologic exam on the cholecystectomy piece. In the areas adjacent to the neoplastic proliferation, premalignant lesions (hyperplasia, metaplasia, dysplasia) were identified in 34.4% cases. The identification of premalignant modifications in the morphologic background of chronic cholecystitis is an argument in favor of the metaplasia-dysplasia-neoplasia sequence and justifies recent recommendations for the performing of colecystectomy.

[Hyperthyroidism and coexistent thyroid carcinoma]. / Hipertiroidismul coexistent cu cancerul tiroidian. Consideratii în legatura cu 11 observatii.

The association between thyrotoxicosis and thyroid cancer is a classical, rare and controversial entity. Within their series of eleven such cases the authors dissociate two distinct groups from clinical and pathological viewpoint. The first category is represented by one case of follicular thyroid cancer with clinically and biologically confirmed hyperthyroidism. The second group includes ten patients with thyrotoxicosis (three cases with Graves' disease, four with toxic multinodular goiter and three with toxic adenoma) and associated unsuspected occult or nodular carcinoma. None of these patients had received previous treatment with radioiodine. In all cases the diagnosis was established by pathological examination of the surgical specimen. Surgery is the treatment of choice in these lesions, the presence of carcinoma imposing the extent of operation. Five near total thyroidectomies and three near total lobectomies with ablation of the isthmus was performed in our cases associated with appropriate additional measures: TSH suppression or 131I treatment. Post surgical results were satisfactory without morbidity or mortality. The long-term survey was good (seven cases alive after 5 years).

[Myocardial injuries induced by oxidative stress in rat--experimental morphopathological study]. / Leziuni miocardice induse de catre stresul oxidativ la sobolan--studiu morfopatologic experimental.

UNLABELLED: Oxidative stress is a phenomenon significantly involved in several clinical conditions such as ischemic heart disease and myocardial reperfusion. PURPOSE: Creating an animal model of oxidative stress based on three methods: exertional (swimming), habitual (light exposure), and corticotropine injections. MATERIAL AND METHODS: 40 Wistar adult rats, weighting between 210-315 g, were subjected to either none (control group), swimming, light exposure or corticotropine injections for six weeks, then the animals were sacrificed and a light and electronic microscopic examination of the myocardium and ascending aorta were performed. RESULTS: Histological features of myocardial reperfusion injury were obtained in all cases and not in the control group. CONCLUSION: All three methods consistently induce myocardial morphological injuries similar to that of the ischemia-reperfusion phenomenon, mainly the exposure to light and corticotropine injections, and mainly in the females.

[Ki-67 and p53 expressions versus differentiation degrees of colon adenocarcinoma]. / Expresia ki-67 si p53 în corelatie cu gradul de diferentiere al adenocarcinomului colonic.

PURPOSE: The colon carcinoma prognostic is evaluated with respect to four categories of factors established by College of American Pathologists Consensus Statement 1999, among which a series of molecular markers is also included. Our study aimed to develop semiquantitative analysis of the immunohistochemistry pattern for proliferation and apoptosis markers, as well as of their connections with the histopathology subtypes of colon adenocarcinoma. MATERIAL AND METHODS: The group of study included 5 cases of well-differentiated adenocarcinoma, 5 cases of moderately differentiated adenocarcinoma and 5 cases of poor differentiated adenocarcinoma. The Ki-67 and p53 antibodies (DAKO) and the streptavidin-biotin method (LSAB Kit, DAKO) were used for the immunohistochemical staining. The semiquantitative analysis provided the Ki-67 and p53 indices, and subsequently their values were statistically correlated with the histologic differentiation degrees. RESULTS: All the cases presented positive immunohistochemistry reaction for Ki-67, the Ki-67 index varying on a large scale, between 5% and 95%. For p53, 10 out of the 15 cases (66%) presented positive immunohistochemistry reaction, with the index value between 22% and 95%, and 5 cases were negative. The statistical analysis revealed no significant difference between the Ki-67 indices of the three histopathologic subtypes (p = 0.674), whereas important differences appeared for the p53 indices (p = 0.007). CONCLUSION: Our findings support the correct understanding of the role played by the immunohistochemical markers, in the sense of the risk involved by considering a pre-definite hierarchy in the diagnosis strategy.

Immunohistochemical markers in the study of gallbladder premalignant lesions and cancer.

AIM: The aim of the study was to perform a comparative analysis of proliferative and apoptotic activity as well as of microvascular density (MVD) in metaplasia, dysplasia and gallbladder carcinoma. MATERIAL AND METHOD: There were investigated 10 cases of chronic cholecystitis associating premalignant lesions and 5 cases of gallbladder carcinoma developed on a chronic cholecystitis background. The immunohistochemical (IHC) staining was made using Ki-67, p53 and CD34 antibodies, and the streptavidin-biotin method. The semiquantitative analysis focused on the intensity of the IHC reaction for Ki-67 and p53 (on a scale from 0 to 4), the Ki-67 index and the percentage of p53 positive cell nuclei. The MVD was established on 5 representative fields as mean of the number of CD34 positive vessels. RESULTS: Ki-67 was positive in all 10 chronic cholecystitis cases (100%) and in all 5 carcinoma cases (100%), which is explainable by the selection manner of chronic cholecystitis associating premalignant lesions and by the invasive pattern of the carcinomas. Ki-67 index rose from 10% in simple dysplastic lesions associated with chronic cholecystitis to 20% in severe dysplastic lesions associated with chronic cholecystitis up until 90% in carcinoma. In all chronic cholecystitis, p53 was negative for normal gallbladder epithelium, in metaplastic territories and in those with mild dysplasia. In the dysplastic areas present in chronic cholecystitis, the positive reaction of p53 rose from 20% (in moderate dysplasia) to 80% (in severe dysplasia). All carcinoma cases were p53 positive (100%), its expression varying from 30% in papillary areas and 30-50% in undifferentiated adenocarcinoma to 80-100% in well differentiated type. In chronic cholecystitis MVD was of 5-9 vessels / field in areas adjacent to pyloric metaplasia and 10-20 vessels / field in severe dysplastic areas. Gallbladder carcinoma cases presented a MVD of 15-30 vessels / field. CONCLUSIONS: The investigation of Ki-67, p53 and MVD brings important data regarding the carcinogenesis process in the gallbladder, initiated on the background of a chronic cholecystitis. The dynamics in the values for Ki-67, p53 and MDV followed from premalignant lesions to carcinoma sustains the hypothesis of the metaplasia-dysplasia-carcinoma sequence.

Computerized morphometric discrimination between normal and tumoral cells in oral smears.

The oral exfoliative cytology allows a quick and fairly accurate assessment of suspicious lesions of the oral cavity. Within this context, our paper proposes a quantitative approach, focusing on the construction of a classifier for detecting the presence of the tumoral cells on oral smears. The design of the classifier relies on a detailed computerized analysis of the individual morphometric features exhibited by two large known populations of normal and tumoral cells, respectively; the digital image processing was performed in the Zeiss KS400 environment. The classifier was implemented as a neural network with step activation function, whose parameters were obtained from an adequate training, based on the nuclear and cytoplasmic areas of the cells belonging to the two populations. Our procedure based on this classifier was meant to operate by identifying the tumoral or normal nature of any cell randomly selected from a smear. To identify the nature of an arbitrary cell, its nuclear and cytoplasmic areas are presented at the input of the classifier. The classification procedure was tested on several smears, and the results coincided with the pathological diagnosis in all the considered cases. The performances of our approach are discussed in comparison with other analytical methods previously reported in oral exfoliative cytology. These discussions emphasize the role of numerical information exploited for the classifier design, concluding that the individual morphometric features are more meaningful than the global characterization of smears by mean values.

[The detection of dysplastic lesions of the gallbladder mucosa in polypoid cholelithiasis]. / Evidentierea leziunilor displazice din mucoasa veziculei biliare în colecistitele calculoase polipoide.

AIM: To investigate the hyperplastic and/or dysplastic lesions in the gallbladder mucosa. METHODS: We used the immunohistochemical Ardeleanu-Hsu-Bussolati and Gugliotta methods, with avidin and peroxide biotin, adapted in the laboratory of University Hospital "St. Spiridon" Iasi. There were studied ten cases of polypoid cholelithiasis, with the following markers: Ki67, PCNA, and p53. RESULTS: The immunohistochemical methods using Ki67, PCNA and p53 protein allowed the evaluation of the abnormal proliferation abilities characteristic for some hyperplastic or dysplastic lesions, found in our cases of cholelithiasis. CONCLUSIONS: The utilization of these markers permitted us to establish a good correlation between the high activity of Ki67 and the morphological aggression indicators in hyperplastic and dysplastic gallbladder epithelium. There is also a correlation between the percentage of the cells marked with anti-PCNA antibody and the existence of p53 protein in these lesions.

[Transcriptional and electron microscopic analysis of extracellular matrix proteoglycans in acute acetaminophen intoxication]. / Analiza la nivel transcriptional si electronomicroscopic a expresiei proteoglicanilor de matrice extracelulara în intoxicatia acuta cu Acetaminofen.

UNLABELLED: Acetaminophen (APAP) OTC regimen is not compatible with kidney and liver toxic effects. In order to further understand the determinism of the repair process, we have studied the expression of some target extracellular matrix proteoglycan components in an animal model. MATERIALS AND METHODS: An acute APAP toxic dose (800 mg/bw) was administered to 45 mice aged 1 month. At 24 hours, tissue samples were processed for light microscopy, electromicroscopy and RT-PCR analysis. RESULTS: Extensive necrotic lesions involving hepatocytes and tubular epithelial cells associate strong positivity for proteoglycans, detected as convoluted filaments with lateral projections, frequently associated with collagen fibers, cell surface and other matrix components. Overexpression for perlecan, syndecan 1 and 4 and low expression for biglycan was assessed by RT-PCR. CONCLUSIONS: Extracellular matrix and cell surface components are early involved in both lesional mechanism and repair reaction post acetaminophen injury.

[Electron microscopy study regarding the etiology and the pathogenesis of acne vulgaris]. / Contributii la studiul electronomicroscopic privind etiopatogenia acneei vulgare.

The etiology and the pathogenesis of acne vulgaris are not complete cleared up yet. This work presents the results of the electronomicroscopic examinations effected on lesions of acne vulgaris biopsied from 5 patients with different clinical forms of disease (including acne fulminans). In this study we had in view the investigation and description of the ultrastructural modifications that followed the intervention of the 4 factors incriminated in the etiology and pathogenesis of acne: sebaceous hypersecretion, hyperkeratosis pilosebaceous infundibulum, bacterial colonisation, perifollicular inflammation. The ultrastructural aspects that we had in view underline the role of the 4 etiology and pathogenesis factors of acne vulgaris, confirming the data related by the specialised literature.

[Pulmonary alveolar proteinosis in AIDS patients]. / Proteinoza alveolara pulmonara la pacientii cu SIDA.

We present 4 patients with pulmonary alveolar proteinosis and AIDS which died in Infectious Disease Hospital Iasi in the interval January 2002-March 2003. The pulmonary tissue from the necropsy exam was stained with H.E., PAS, Alcian blue and Mucicarmine. The macroscopical examination revealed multiple yellow-gray nodular areas (>2 cm) of consolidation. Microscopically, the alveoli were filled with a delicate, granular, acidophil material that was periodic acid Schiff (PAS) positive. The intraalveolar material was Alcian blue and Mucicarmine negative, too.

[Electron microscopy in oral squamous cell carcinoma of elderly patients]. / Aspecte electronomicroscopice în carcinoamele cavitatii orale la pacientii de vârsta a treia.

OBJECTIVES: Squamous cell carcinoma is the most frequent tumor occurring in the oral cavity. The aim of the study is to point out the fine-structural characteristics of the tumor cells and of the adjacent stromal cells, trying to establish a correlation between these characteristics and the biological behavior of the tumor. MATERIAL AND METHODS: We performed an electron microscopic investigation for tree elderly patients diagnosed with oral squamous cell carcinoma. RESULTS: The tumoral cells presented nuclear abnormalities and the reduction of the intercellular cohesion, through an increase in the intercellular spaces, and desmosomal modifications. Through the abundance and the variety of the cellular types, the stromal component represented a positive response from the host. CONCLUSIONS: In the diagnosis of the oral carcinoma, the electron microscopic exam supplements the light microscopic investigation, revealing important morphologic elements at a subcellular level (regarding the nucleus, cellular organelles, and junctions).