Comparison of uroplakin expression during urothelial carcinogenesis induced by N-butyl-N-(4-hydroxybutyl)nitrosamine in rats and mice

The expression of uroplakins, the tissue-specific and differentiation- dependent membrane proteins of the urothelium, was analyzed immunohistochemically in N butyl-N-(4-hydroxybutyl)nitrosamine (BBN)-treated rats and mice during bladder carcinogenesis. Male Fischer 344 rats were treated with 0.05% BBN in the drinking water for 10 wk and were cuthanatized at week 20 of the experiment. BBN was administered to male B6D2F1 mice; it was either provided at a rate of 0.05% in the drinking water (for 26 wk) or 5 mg BBN was administered by intragastric gavage twice weekly for 10 wk, followed by 20 wk without treatment. In rats, BBN-induced, noninvasive, low grade, papillary, transitional cell carcinoma (TCC) showed decreased uroplakin-staining of cells lining the lumen but showed increased expression in some nonluminal cells. In mice, nonpapillary, high-grade dysplasia, carcinoma in situ, and invasive carcinoma were induced. There was a marked decrease in the number of uroplakin-positive cells lining the lumen and in nonluminal cells. This occurred in normal-appearing urothelium in BBN-treated mice and in dysplasic urothelium, in carcinoma in situ, and in invasive TCC. The percentage of uroplakin-positive nonluminal cells was higher in control mice than in rats, but it was lower in the mouse than in the rat after BBN treatment. Uroplakin expression was disorderly and focal in BBN-treated urothelium in both species. These results indicate that BBN treatment changed the expression of uroplakins during bladder carcinogenesis, with differences in rats and mice being related to degree of tumor differentiation.

Comparative experimental infection of Lacazia loboi in BALB/c and B10.A mice

Both hind foot pads of BALB/c and B10.A mice strains, were inoculated with a fungal suspension of Lacazia loboi obtained from a Jorge Lobo's disease patient. The suspension had 9 × 105 cells/ml and its viability index was 45%. The animals were sacrificed at different time periods varying from 24 h to 18 months after inoculation. The BALB/c mice developed an extensive granulomatous infiltrate, similar to the disease in humans, that progressively evolved. The number of fungal elements also increased as the disease progressed, and after the seventh month of inoculation, macroscopic changes of the foot pads were evident. Although the B10.A mice developed an exuberant granulomatous infiltrate, macroscopic changes were not detected. The number of fungal cells in the infected tissues increased in number, but they were lower then the numbers found in the BALB/c strain. The viability indexes were also lower for the B10.A strain. Considering the histopathological findings, the presence of macroscopic changes and the great amount of fungal cells in the infected tissues, the authors concluded that the BALB/c mice strain was more susceptible to L. loboi infection than the B10.A strain.

Effect of air-powder system on titanium surface on fibroblast adhesion and morphology.

PURPOSE: To evaluate the number and morphology of fibroblasts grown on machined titanium healing abutments treated with an airpowder system. MATERIALS AND METHODS: Twenty-six abutments were assigned to two experimental groups: control (no treatment) and treated (exposed to the Prophy-Jet for 30 seconds). The specimens were incubated for 24 hours with fibroblastic cells in multiwell plates, followed by routine laboratory processing for scanning electron microscope analysis. The specimens were photographed at x 350, and the cell number was counted on an area of approximately 200 um2. RESULTS: No significant differences were found on morphology between the groups (P > 0.05); however, the control group presented a significantly greater amount of cells (71.44 +/- 31.93, mean +/- SD) in comparison with treated group (35.31 +/- 28.14), as indicated by a nonpaired t test (P = 0.001). CONCLUSION: The use of an air-abrasive prophylaxis system on the surface of titanium healing abutments reduced the cells proliferation but did not influence cell morphology.

Características hematológicas, bioquímicas e biométricas de Piaractus mesopotamicus Holmberg, 1887 (Osteichthyes: Characidae) oriundos de cultivo intensivo

Hematological characteristics are important tools for diagnosis and prognostic features in fish population and they contribute to the comprehension of comparative physiology, phylogenetic relation, food habit and other ecological parameters. Red blood cell count (RBC), white blood cell count (WBC), thrombocytes counts, glucose, cortisol, hepatosomatic and splenosomatic relation were evaluated in Piaractus mesopotamicus. The values of hepatosomatic relation varied from 0.67% to 2.79% and splenosomatic relation from 0.011% to 0.108%. RBC 1.870 to 4.590 × 106/μL, hemoglobin concentration 5.7 to 12.1g/dL, haematocrit 24.0% to 40.0%, mean corpuscular volume 98.0 to 174.0 fL, mean corpuscular haemoglobin concentration 19.1 to 34.2g/dL, glucose 40.6 to 89.2mg/dL and cortisol 4.73 to 110.5ng/dL The number of total thrombocytes was 12,160.0 to 89,280.0/μL and WBC 2,019.0 to 47,473.0/μL. Differential count of leucocytes showed 54.1% limphocytes, 10.9% neutrophils, 10.3% monocytes, 9.9% PAS-positive granular leukocytes (PAS-GL), 6.7% eosinophils, 0.3% basophils and 7.8% immature leucocytes. Variation of these haematological characteristics in the same fish species was observed, being considered as the normal variation in the fish concerned. Hence it is incorrect to assume that biological characteristics in fishes are distributed normally.

Effects of long-term cyclosporin therapy on the periodontium of rats

Background: The treatment of cyclosporin A triggers an early bone loss and gingival overgrowth. There is a lack of studies exploring the effects of long-term cyclosporin A therapy on alveolar bone homeostasis and gingival tissue. Objective: The purpose of this study was to evaluate the effects of long-term therapy with cyclosporin A on the gingival tissue and on the alveolar bone metabolism in rats. Materials and methods: Rats were treated for 60, 120, 180 and 240 days with a daily subcutaneous injection of 10 mg/kg body weight of cyclosporin A. At the end of experimental periods, animals were killed and the serum calcium (Ca2+) and alkaline phosphatase levels were measured in all groups. After histological processing, the oral epithelium and the connective tissue, as well as volume densities of alveolar bone (Vb) and multinucleated osteoclasts (Vo), were assessed at the region of the lower first molars. Results: Significant increases in the serum alkaline phosphatase were observed in those groups that received cyclosporin A therapy. After 60 and 120 days of the treatment with cyclosporin A, evident gingival overgrowth associated with a significant increase of epithelium and connective tissue was observed, as well as a decrease of the densities of bone and an increase of densities of osteoclasts. After 180 and 240 days of the treatment, there was a reduction of the gingival overgrowth associated with significant decreases of epithelium and connective tissue, as well as an increase of bone densities and a decrease of osteoclasts. Conclusion: Within the limits of this experimental study, it can be concluded that the deleterious periodontal effects of cyclosporin A administration may be time-related side-effects. © Blackwell Munksgaard, 2004.

Plication of the free wall of the left ventricle in dogs with doxorubicin-induced cardiomyopathy

Objective - To evaluate plication of the free wall of the left ventricle, which reduces the left ventricular area and volume, as a method to improve the left ventricular systolic function without cardiopulmonary bypass. Animals - 8 mixed-breed adult dogs. Procedure - Dilated cardiomyopathy (DCM) was induced in each dog by administration of doxorubicin (30 mg/m2, IV, q 21 d for 168 days). Two dogs died during induction of cardiomyopathy. Plication surgery was performed in 4 dogs. Two dogs did not ondergo to surgery (control group). Values for cardiac output (CO), 2-dimensional and M-mode echocardiography, arterial blood pressure, electrocardiography, blood cell counts, and serum biochemical analyses were recorded after induction of DCM (baseline) and 1, 2, 7, 15, 21, 30, 60, 90, 120, 150, and 180 days after plication surgery. Ambulatory ECG (Holter) recordings were conducted for 24 hours on the day of surgery. Results - 1 dog died after plication surgery. The remaining dogs undergoing ventricular plication had a significant improvement in CO, ejection fraction, and fractional shortening and reductions of left ventricular area and volume after surgery. Electrocardiographic and Holter recordings revealed premature ventricular complexes, which resolved without treatment during the first week after surgery. Clinical condition of the control dogs declined, and these 2 dogs died approximately 40 days after induction of cardiomyopathy. Conclusions and Clinical Relevance - Plication of the free wall of the left ventricle improved left ventricular systolic function in dogs with doxorubicin-induced cardiomyopathy. Additional studies are needed to evaluate its application in dogs with naturally developing DCM.

Effects of long-term cyclosporin therapy on gingiva of rats--analysis by stereological and biochemical estimation.

Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of gingival overgrowth, which remains a significant problem. The risk factors appraised include the duration of treatment. However, there are no stereological and biochemical studies exploring the effects of long-term CsA therapy on gingival tissue. The purpose of the present study was to investigate the level of TGF-beta1 in saliva and describe the densities of fibroblasts and collagen fibers in the gingival tissue of rats treated with CsA for long periods. Rats were treated for 60, 120, 180 and 240 days with a daily subcutaneous injection of 10 mg/kg of body weight of CsA. At the end of the experimental periods, saliva was collected for the determination of TGF-beta1 levels. After histological processing, the oral epithelium and the connective tissue area were measured as well as the volume densities of fibroblasts (Vf) and collagen fibers (Vcf). After 60 and 120 days of CsA treatment, there was a significant increase in Vf and Vcf as well as a significant increase in TGF-beta1 levels. After 180 and 240 days, reduction in the gingival overgrowth associated with significant decreases in the level of TGF-beta1, and also decreased Vf and Vcf, were observed. The data presented here suggest that after long-term therapy, a decrease in TGF-beta1 levels occurs, which might contribute to an increase in the proteolytic activity of fibroblasts in the gingiva, favoring the normality of extracellular matrix synthesis.

Morphometric and quantitative evaluation of the NADH-diaphorase positive myenteric neurons of the jejunum of streptozotocin-diabetic rats supplemented with acetyl-L-carnitine

In this study we investigated the effect of the acetyl-L-carnitine (ALC) supplementation on the myenteric neurons of the jejunum of rats made diabetic at the age of 105 days by streptozotocin (35 mg/kg body weight). Four groups were used: non-diabetic (C), non-diabetic supplemented with ALC (CC), diabetic (D), diabetic supplemented with ALC (DC). After 15 weeks of diabetes induction the blood was collected by cardiac puncture to evaluate glycaemia and glycated haemoglobin. Next the animals were killed and the jejunum was collected and subjected to whole-mount preparation to evidence the myenteric neurons through the histochemical technique of the NADH-diaphorase. The neuronal counts were made in 80 microscopic fields, in tissue samples of five animals of each group. The profiles of the cell bodies of 1000 neurons per group were analysed. Diabetes induced a significant increase in the area of the cell body and decrease in the number of NADH-diaphorase positive myoenteric neurons. ALC suplementation to the diabetic group promoted smaller hypertrophic effects and less neuronal loss than in the myoenteric neurons of the diabetic rats, and in addition diminished the body weight decrease and reduced the fasting glycaemia. © 2005 Blackwell Verlag.

The number of podocyte and slit diaphragm is decreased in experimental diabetic nephropathy

Purpose: To determine the number of podocyte, slit diaphragms, slit diaphragm extensions and GBM thickness in diabetic nephropathy. Methods: Sixty Rattus Wistarof both sexes weighing 200-300g were divided in two experimental groups: normal group 10 animals, and alloxan diabetic rats - 50 animals. Alloxan was administered in a single IV dose of 42mg/kg body weight. Body weight, water and food intake, diuresis, and blood and urine glucose were determined in both groups before alloxan injection and two weeks, six and twelve months after alloxan injection. Proteinuria was measured at 12 months in both groups. After 12 months animals were sacrificed, and the right kidney processed for electron microscopy. Results: Clear clinical and laboratory signs of severe diabetes were seen, in all alloxan-diabetic rats at all follow-up times. Glomerular basement membrane (GBM) thickening, podocyte number, and slit diaphragm number and extension were determined. GBM of all diabetic rats was significantly thicker (median=0.29μm; semi-interquartile range=0.065μm) than in the normal rats (0.23μm; 0.035μm). Diabetic rat podocyte number (8; 1), slit diaphragm number (4; 1), and slit diaphragm extension (0.021μm; 0.00435μm) were significantly lower than in normal rats (11; 1) and (7; 1.5), and (0.031μm; 0.0058μm). Diabetic rat proteinuria (0.060mg/24h; 0.037mg/24h) was higher than in normal rats (0.00185mg/24h; 0.00055mg/24h). Conclusion: Experimental diabetes is associated with significant (p<0.05) changes in podocyte foot process, slit number, slit diaphragm extension, and GBM thickness.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD 45RA and CD 45RO cell count; determination of IL-2, IFN-γ, IL-4, IL-10 and TNF-α serum levels; CD 4 + T and CD 8 + T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD 45RA (G1<[G3=GC-2]; p<0.05) and CD 45RO (G1cells, and CD 4 + T lymphocyte count (G1[G2=G3]; p<0.001), TNF-α serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-γ ([G1=GC-1]>[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD 8 + T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-γ), CD 45RA and CD 45RO cells, and CD 4 + T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-α, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-γ, and VL, followed by CD 45RA and CD 45RO cells, and IL-10. The variables with the lowest number of correlations were CD 4 + T and CD 8 + T lymphocytes. The results express the partial but important immune reconstitution in HIV-1 infected individuals with the interference of HAART and the importance of cytokines especially IL-2 and IFN-γ, and CD 45RA and CD 45RO cells as surrogate markers of this reconstitution.

Morphometric study of the postnatal growth of the parotid gland of the mouse

The growth of the mouse parotid glands during 7 and 35 days of postnatal life was studied by morphometric methods. The mass of the gland, the volume of each morphological compartment, and the cell number in each compartment were evaluated. The data obtained for each evaluated dimension were adjusted by an exponential equation, of the type Y = a.e KX, thus permitting the calculation of their mean duplication time (T D), i.e., an estimation of their growth rate. Analysis of the results showed a marked 1,424% increase in the gland mass during the whole studied period, with T D = 7.10 days. This growth occurred by increases in absolute volume of acini, intercalated ducts, striated ducts, excretory ducts and stroma, with percentual increases of 3,048%, 417%, 2,662%, 2,594% and 367%, respectively, and T Ds of 5.62, 11.71, 5.55, 5.47 and 14.45 days, respectively. Analysis of the cell number growth in each compartment showed increases of 1,904%, 285%, 1,228%, 1,090% and 286%, respectively, and T Ds of 6.62, 20.40, 7.19, 7.26 and 14.51 days, respectively. Based on the present results, we concluded that the growth of the mouse parotid glands from day 7 to day 35 of age occurred by intense cell accumulation, mainly in the acini, striated ducts and excretory ducts, with a growth rate sensibly higher than that of the intercalated ducts and stroma.

Levedura íntegra e derivados do seu processamento em rações para tilápia do Nilo: Aspectos hematológicos e histológicos

The effects of inclusion of whole yeast, autolyzed yeast and yeast cell wall on hematological parameters and gut villus perimeter were evaluated in juvenile Nile tilapia, after 80 experimental days. Isoproteic (32.0% DP) and isoenergetic (3200 kcal DE kg -1) practical diets were supplemented with three levels of whole yeast or autolyzed yeast (1.0, 2.0 and 3.0%) and three levels of yeast cell wall (0.1, 0.2 and 0.3%), plus a control diet (with no test microingredients). Red blood cell count, hemoglobin concentration, total plasmatic protein, hematocrit percentage, mean corpuscular volume, mean corpuscular hemoglobin concentration and gut villus perimeter were evaluated. Variations on hematological parameters in animals fed diets with whole yeast; autolyzed yeast and yeast cell wall were observed to be within normal ranges for this species. There was significant influence (p<0.05) of different levels of yeast and derivatives on intestinal villus perimeter. Results showed that experimental period and proposed levels of whole yeast, autolyzed yeast and yeast cell wall do not provide undesirable alterations on standard hematological parameters to Nile tilapia and can be safely used to compound diets for this species. Results also showed that supplement of yeast cell wall provide higher intestinal villus perimeter.

Recombinant LH supplementation to recombinant FSH during induced ovarian stimulation in the GnRH-antagonist protocol: A meta-analysis

This study aims to compare the efficacy of recombinant LH (rLH) supplementation for ovarian stimulation in gonadotrophin-releasing hormone-antagonist protocol for IVF/intracytoplasmic sperm injection cycles. Search strategies included online surveys of databases. The fixed effects model was used for odds ratio (OR) and effect size (weighted mean difference, WMD). Five trials fulfilled the inclusion criteria. When the meta-analysis was carried out, advantages were observed for the LH supplementation protocol with respect to higher serum oestradiol concentrations on the day of human chorionic gonadotrophin administration (P < 0.0001; WMD: 514, 95% CI 368, 660) and higher number of mature oocytes (P = 0.0098; WMD: 0.88, 95% CI 0.21, 1.54). However, these differences were not observed in the total amount of recombinant FSH (rFSH) administered, days of stimulation, number of oocytes retrieved, the clinical pregnancy rate per oocyte retrieval, the implantation rate and miscarriage rate. This result demonstrates that the association of rLH with rFSH may prevent any decrease in oestradiol after antagonist administration and that a significantly higher number of mature oocytes was available for laboratory work. Nevertheless, it failed to show any statistically significant difference in clinically significant end-points in IVF (implantation and pregnancy rates). Additional randomized controlled trials are needed to confirm these results further. © 2007 Published by Reproductive Healthcare Ltd.

Antisperm antibodies in infertile men and their correlation with seminal parameters

Aim: Antisperm antibodies (ASA) in males cause the autoimmune disease 'immune infertility'. The present study intended to detect the presence of ASA and their incidence in men with unexplained infertility, as well as to evaluate the correlation between the presence of ASA and semen parameter alterations. Methods: Blood and sperm assessment were collected to carry out a direct and indirect mixed antiglobulin reaction (MAR) test and semen analysis in infertile and fertile men from the University Hospital of the Faculty of Medicine, Sao Paulo State University, Sao Paulo. Results: In the MARtest, 18.18% of infertile men were positive for ASA. In fertile men, no positivity was found. A significant correlation between the presence of ASA with an increased white blood cell count plus a decreased hypoosmotic swelling test result was observed. Conclusions: The results indicate that ASA are involved in reduced fertility. It is not ASA detection per.se that provides conclusive information about the occurrence of damage to fertility. The correlation between infertility and altered seminal parameters reinforce the ASA participation in this pathology. © 2007 The Authors Journal compilation. © 2007 Japan Society for Reproductive Medicine.

Evaluation of adverse effects of long-term oral administration of carprofen, etodolac, flunixin meglumine, ketoprofen, and meloxicam in dogs

Objective - To evaluate adverse effects of long-term oral administration of carprofen, etodolac, flunixin meglumine, ketoprofen, and meloxicam in dogs. Animals - 36 adult dogs. Procedures - Values for CBC, urinalysis, serum biochemical urinalyses, and occult blood in feces were investigated before and 7, 30, 60, and 90 days after daily oral administration (n = 6 dogs/group) of lactose (1 mg/kg, control treatment), etodolac (15 mg/kg), meloxicam (0.1 mg/kg), carprofen (4 mg/kg), and ketoprofen (2 mg/kg for 4 days, followed by 1 mg/kg daily thereafter) or flunixin (1 mg/kg for 3 days, with 4-day intervals). Gastroscopy was performed before and after the end of treatment. Results - For serum γ-glutamyltransferase activity, values were significantly increased at day 30 in dogs treated with lactose, etodolac, and meloxicam within groups. Bleeding time was significantly increased in dogs treated with carprofen at 30 and 90 days, compared with baseline. At 7 days, bleeding time was significantly longer in dogs treated with meloxicam, ketoprofen, and flunixin, compared with control dogs. Clotting time increased significantly in all groups except those treated with etodolac. At day 90, clotting time was significantly shorter in flunixin-treated dogs, compared with lactose-treated dogs. Gastric lesions were detected in all dogs treated with etodolac, ketoprofen, and flunixin, and 1 of 6 treated with carprofen. Conclusions and Clinical Relevance - Carprofen induced the lowest frequency of gastrointestinal adverse effects, followed by meloxicam. Monitoring for adverse effects should be considered when nonsteroidal anti-inflammatory drugs are used to treat dogs with chronic pain.