Computer-assisted analysis of cell proliferation markers in oral lesions

Abnormalities in any component of the cell cycle regulatory machine may result in oral. cancer, and markers of cell proliferation have been used to determine the prognosis of tumor progression. The aim of this study was to determine whether silver-stained nucleolar organizer region (AgNOR) and Ki-67 measurements could improve the assessment of growth rates in oral lesions. Eighty-three oral biopsies were studied, 20 of which were classified as fibrous inflammatory hyperplasia (FIH), 40 as leukoplakia (LKP) and 23 as oral. squamous cell carcinoma (OSCC). Within the LKP group, 22 out of 29 biopsies were diagnosed as non-dysplastic leukoplakia (LK) and 18 as dysplastic teukoptakia (DLK), presenting discrete, moderate and severe dysplasia. Ki-67 immunotabeting of the lesions increased steadily in the following order: FIH, DLK, LK and OSCC, indicating that Ki-67 is a good marker for predicting the protiferative fraction among benign, premalignant and malignant oral lesions. The median values of AgNOR parameters indicate that the morphometric index gives better results regarding the proliferative rate than the numerical one. A series of linear regressions between AgNOR parameters and Ki-67 showed positive associations. We conclude that a combination of Ki-67 and morphometric AgNOR analyses could be used as an aid in the determination of the protiferative status of oral epithelial. cells in oral cancer. (C) 2007 Elsevier GmbH. All rights reserved.

EFFECT OF ASCORBIC-ACID DEFICIENCY ON THE GROWTH, GILL FILAMENT LESIONS AND BEHAVIOR OF PACU FRY (PIARACTUS-MESOPOTAMICUS HOLMBERG, 1887)

The effect of ascorbic acid deficiency was determined in Piaractus mesopotamicus Holmberg, 1887, fish (pacu) under laboratory conditions. A total of 120 fish with an average body weight of 8.64 +/- 1.62 g and measuring 6.15 +/- 0.33 cm in length at the beginning of the experiment were fed diets containing 0, 50, 100 or 200 mg palmitate-coated ascorbic acid/kg dry ration for a period of 24 weeks with measurements every 4 weeks. The experiment was conducted in 20 fiber-cement aquaria of 81-liter capacity. Each aquarium was supplied with dechlorinated water at a flow rate of 1 l/min. Water temperature was measured daily and pH, dissolved oxygen, alkalinity and water conductivity were determined weekly. A fully randomized experimental design was utilized, with 5 replicates of each treatment and 6 fish per aquarium. Ascorbic acid-supplemented fish presented significantly increased growth when compared to unsupplemented fish. Furthermore, unsupplemented fish presented a higher incidence of hyperplasia, hypertrophy and dysplasia of the bone cartilage of gill filaments. The gill lamellae of unsupplemented fish had twisted cartilage and an inflammatory infiltrate at the ends. Anorexia and increased handling stress were also observed in fish fed the unsupplemented diet. The present study suggests that 50 mg ascorbic acid/kg dry ration is sufficient to improve development of pacu fingerlings but the optimum level under aquarium conditions, determined by regression analysis, is 139 mg ascorbic acid/kg dry ration.

P53 PROTEIN EXPRESSION AND NUCLEAR-DNA CONTENT IN BREAST INTRADUCTAL PROLIFERATIONS

Immunohistochemical analysis of the p53 gene protein and cytometric assessment of nuclear DNA were performed in a series of 51 cases of intraductal breast proliferation. The series included 22 cases of intraductal hyperplasia without atypia, 6 cases of intraductal hyperplasia with atypia, and 23 cases of pure intraductal carcinoma. Expression of p53 protein was detected in one case of intraductal hyperplasia without atypia (4.5 per cent), one case of intraductal hyperplasia with atypia (16.6 per cent) and six cases of intraductal carcinoma (26.0 per cent). No significant correlation was observed between p53 expression and histological subtype of intraductal carcinoma. Aneuploidy was demonstrated in two cases of intraductal hyperplasia with atypia (33.3 per cent) and in 18 cases of intraductal carcinoma (78.2 per cent). All cases of intraductal hyperplasia without atypia were euploid. No significant association was observed between p53 protein expression and ploidy in intraductal hyperplasia. The only case of intraductal hyperplasia without atypia positive for p53 was euploid, whereas the only p53-positive case of intraductal hyperplasia with atypia was aneuploid. Among the intraductal carcinomas, only the aneuploid cases showed positivity for p53, regardless of histological subtype. The results suggest that some of the changes observed in invasive breast carcinoma, such as p53 expression and aneuploidy, are already present in breast intraductal proliferation, especially in areas with atypia and in intraductal carcinoma. The expression of p53 in breast intraductal proliferation may reflect the acquisition of p53 gene mutations in cells unable adequately to repair DNA damage, with genomic instability which would lead to clonal expansion and putative evolution to invasive disease.

Tissue remodeling in Guinea pig lateral prostate at different ages after estradiol treatment

Estrogen seems to have an essential role in the fibromuscular growth characteristic of benign prostatic hyperplasia (BPH). This paper describes the effects of chronic estradiol treatment on Guinea pig prostatic stroma at different ages. Tissues from experimental animals were studied by histological and histochemical procedures, morphometric-stereological analysis and transmission electron microscopy (TEM). Marked fibromuscular hypertrophy was observed after estradiol treatment in animals of pre-pubertal and adult ages. Increases in the density and thickness of the collagen and elastic fibers were observed by histochemistry. TEM revealed wide distributions of collagen fibrils and large elastic fibers adjacent to the epithelial basal lamina and between the stromal cells, establishing contacts between them. These results indicate that the Guinea pig prostate simulates the stromal modifications observed in BPH in some aged animals after estrogen treatment at different ages, making it a good model for this disease. (c) 2005 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Spectrum of carcinoembryonic antigen immunoreactivity from isolated ductal hyperplasias to atypical hyperplasias associated with infiltrating ductal breast cancer

Aims - To study the immunohistochemical expression of carcinoembryonic antigen (CEA) in ductal hyperplasia of the breast and to investigate its putative relation with atypia and co-existing infiltrating ductal carcinoma.Methods - Paraffin wax embedded tissue from 37 cases of isolated ductal hyperplasia (five with atypia and 32 without atypia) and 25 cases of ductal hyperplasia associated infiltrating ductal carcinoma (IDC) (seven with atypia and 18 without atypia) was stained with a monoclonal anti-CEA antibody using a standard avidin biotin immunoperoxidase method.Results - CEA immunoreactivity was observed in eight (12.8%) ductal hyperplasia cases. The percentage of CEA positivity in ductal hyperplasia cases with atypia (33.3%) was substantially higher than that observed in cases of ductal hyperplasia without atypia (8.0%). Six cases of ductal hyperplasia associated IDC reacted with CEA; in these six cases the neoplastic cells of the co-existing carcinoma were also CEA positive. The percentage of CEA immunoreactivity in ductal hyperplasia associated IDC was higher than that observed in isolated ductal hyperplasia (24.0 v 5.4%). The percentage of CEA immunoreactivity in atypical ductal hyperplasia associated IDC was similar to that observed in IDC alone (42.9 v 40.0%).Conclusions-The presence of CEA immunoreactivity has been confirmed in benign proliferative breast lesions. The prevalence of such immunoreactivity increases from 3.1% in isolated, nonatypical ductal hyperplasia to 42.9% in atypical ductal hyperplasia associated IDC. This finding and the similarity of the frequency of CEA positivity in atypical ductal hyperplasia associated IDC and in IDC alone suggests that there is a pathogenetic link between ductal hyperplasia and some types of breast cancer.

Immunohematopathology markers in paraffin sections

New monoclonal antibodies have made the use of formalin fixed, paraffin embedded sections practical for immunotyping and immunodiagnosing many hematolymphoid neoplasms. In addition, they can be helpful in classifying malignant lymphomas and leukemias through patterns of immunoreactivity. The most useful. lymphoid antibodies and diagnostic situations in which they can be deployed are discussed.

Experimental infectious bursal disease in the ostrich (Struthio camelus)

Clinically severe disease was produced in ostriches aged 4 weeks by oral infection with avirulent strain of infectious bursal disease virus (vIBDV), namely strain Faragher 52/70. Four days after infection the birds were humanely killed and tissue samples, including thymus, bursa of Fabricius (BF), brain and kidney were collected for examination. Histopathologically, the thymus and BF showed severe lymphoid depletion and necrosis, while immunolabelling with a polyclonal antibody demonstrated abundant viral antigen. (C) 2007 Elsevier Ltd. All rights reserved.

Collagen type VI is a component of the extracellular matrix microfibril network of the prostatic stroma

Stroma-epithelium relationships are of great relevance in prostatic morphogenesis and physiology, However, little knowledge exists about either stromal cells or extracellular matrix composition and arrangement in this system, Ultrastructural analysis revealed the existence of a microfibrillar system which occupies large areas of the rat prostatic stroma, In this work, we have applied immunocytochemistry and an ATP treatment for the ultrastructural identification of collagen type VI microfibrils, aiming at examining its participation in the prostatic microfibrillar network. Immunocytochemistry was also extended to a human case of prostatic nodular hyperplasia, Both methods succeeded in identifying collagen type VI in the rat ventral prostate, Collagen type VI is evenly distributed throughout the stroma but mainly associated with the basal lamina, collagen fibrils, and around the stromal cells, the use of ATP treatment allowed for the discrimination between collagen type VI and elastin-associated microfibrils, and demonstrated that these two classes of microfibrils establish an extended, mixed, and open network. The same aspects of association with the basal lamina, with stromal cells (particularly with smooth muscle cells), and with fibrillar components of the stroma were observed in the human tissue, We suggest that the collagen type VI and elastin-associated microfibril system may be involved in the control of some aspects of cellular behavior and may also play a structural role, maintaining the organ integrity after the deformations occurring under smooth muscle contraction.

Estudo experimental sobre a intoxicação de Gallus gallus domesticus com semente de crotalaria spectabilis. II efeito em aves na fase final de crescimento

This study was undertaken to investigate the toxic effect of Crotalaria spectabilis seeds added to the ration for commercial broilers during the final phase of growth. Ground seeds were added at different concentrations to the ration: 0.0% (control), 0.01%, 0.1% and 0.4%. Rations containing 0.4% caused symptoms of intoxication, beginning in the second week of the study. During the third week birds showed bristling, apathy, general weakness, distended abdomen and agglomeration. In the fourth week, four animals died. Necropsy revealed prominent ascites and severe lesions of liver, kidney and lung. Microscopic examination revealed necrosis of hepatocytes, inflammatory cell infiltration, hypertrophy and hyperplasia of biliary duct cells leading to atresia and cholestasis. Prominent cartilaginous and osseous nodules in the lungs were also present as well as degenerative, changes in the kidney tubules and necrosis of cells of the bursa. All the broilers receiving a ration with 0.1% of Crotalaria seeds showed ascites and slight lesions of the liver. The remaining groups showed no reduction in weight gains, lesions or clinical symptoms. It is concluded that broilers during the final phase of growth are sensitive to administrations of seeds of C. spectabilis in their ration. The presence of ascites and cartilaginous and osseous nodules in the lungs of the affected birds was also considered important.

REDUCTION OF CELL PROLIFERATIVE ACTIVITIES OF GASTRIC STUMP ADENOMATOUS HYPERPLASIAS AFTER BILE REFLUX DIVERSION IN RATS

Previously we reported the majority of lesions induced hy bile reflux, in the absence of chemical carcinogens, in the rat remnant stomach to consist primarily of gastric type and secondarily of intestinal type cells, and that they are reversible after diversion of bile reflux. The present study was designed to evaluate changes in proliferative activities in cells of each type under these conditions. The frequency of adenomatous hyperplasia (AH) induced in the gastric stump mucosa by duodenal content reflux after Billroth II partial gastrectomy (BII) increased until the 54th week of the experiment. Roux-en-Y (RY) surgical procedure which prevents duodenal reflux performed at the 24th or 36th week after BII led to a decrease in AH. Cell content of the lesions was analyzed using routine H&E staining, immunohistochemical staining for pepsinogen isoenzyme 1 and histochemical procedures for mucins (paradoxical concanavalin A, galactose oxidase Schiff and sialidase galactose oxidase Schiff reactions) and proliferation in each compartment evaluated by an immunohistochemical method using bromodeoxyuridine (BrdU) and a monoclonal antibody against BrdU. At the 54th week the number of BrdU-labeled cells per normal pyloric column was significantly (P < 0.05) increased to 10.63/pit after the BII operation, while it diminished to 5.23/pit after RY diversion, this being the same level as with the RY procedure alone. AH maintained a high rate of BrdU incorporation at 12.7% after BII operation, which was also significantly reduced (P < 0.01) to 7.0% by the RY surgery. The intestinal type cell showed highest (22.2%), the surface mucous type cell showed the next (16.5%) and the pyloric gland type cell showed lowest (5.2%) BrdU labeling indices after BII operation. All the cell types in AH showed similar proportional decreases in BrdU incorporation after RY diversion. Thus surgical intervention reverses the cell proliferation caused by bile reflux in the gastric stump.

Mummified papyraceous fetuses in the abdominal cavity of an elderly female dog with pyometra

This paper reports on a rare case of fetal papyraceous mummification after asymptomatic uterine rupture in an elderly female dog with pyometra. The patient had a history of mating six months before the examination but no apparent signs of gestation or parturition. Exploratory laparotomy was used to identify a rupture of the left uterine horn and the presence of cystic endometrial hyperplasia and pyometra. Two mummified papyraceous fetuses were observed in the abdominal cavity and had adhered to the spleen, pancreas, intestine and omentum. Ovariehysterectomy and corrective surgery were performed. The patient had remained healthy after uterine rupture until a new estrous cycle and the development of pyometra. Bitches that are 10 years old or more are predisposed to implantation failure, pregnancy or parturition problems and they should not be breed to avoid complications.

Maternal antibody transfer to broiler progeny varies among strains and is affected by grain source and cage density

Two experiments were conducted to examine the effects of broiler breeder dietary grain source and cage density on maternal antibody (MatAb) transfer to progeny in 2 genetic strains (A and B). Broiler breeders were assigned to 16 litter floor pens and fed either corn- or wheat-based diets. Breeders were administered 4 live vaccines against Newcastle disease virus (NDV). At 23 wk of age, pullets and cocks, which reflected the full BW distribution from each treatment, were moved to a cage breeder house and placed at 1 or 2 hens/cage. Breeders were artificially inseminated at 44 wk (experiment 1) and 52 wk of age (experiment 2). Eggs were collected for 8 d, incubated, and placed in individual pedigree bags at d 19 of incubation. Blood samples from 5 chicks per treatment combination were collected at hatch in both experiments. Spleen and bursa were collected from the same chicks for histomorphometry analyses in experiment 2. In the second experiment, 12 chicks per treatment were placed in cages. Progeny were provided diets based on the same grain (corn or wheat) as their parents. Serum samples were collected at 5, 9, and 13 d of age and analyzed for anti-NDV MatAb. Data were analyzed as a 2 x 2 x 2 factorial design considering strain, dietary grain source, and cage density as main factors. Interaction effects were observed in breeders and progeny. Experiment 1 showed that strain A chicks had lower levels of MatAb when hens were housed at 2 hens/cage rather than 1 hen/cage. The MatAb levels of strain B chickens were not affected by cage density in either experiment. Experiment 2 demonstrated similar effects of cage density on MatAb levels and the area of bursa follicles for both strains. Progeny of breeders fed corn-based diets had smaller spleen white pulp only when hens were housed at 2 hens/cage compared with 1 hen/cage. The results of these experiments suggest that breeder strain and cage-density conditions affected MatAb transfer to progeny and embryo development of spleen and bursa.

Leishmania chagasi infection in cats with dermatologic lesions from an endemic area of visceral leishmaniosis in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Leishmania chagasi infection in cats with dermatologic lesions from an endemic area of visceral leishmaniosis in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High doses of dexamethasone induce increased beta-cell proliferation in pancreatic rat islets

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)