Estudo da toxicidade de diferentes doses do Diuron (3-(3,4 -dichlorophenyl)-1,1-dimethylurea) no epitélio da bexiga de ratos wistas machos por microscopia eletrônica de varredura

Diuron (3-(3,4-Dichlorophenyl)-1,1-dimethylurea) is a substituted urea herbicide widely used on agricultural crops such as soy, cotton and sugar cane. In a previous long-term study this herbicide exerted carcinogenic activity on the urinary bladder and renal pelvis mucosa of Wistar rats and breast of mice. Also, it was shown to be carcinogenic to the mice skin in a initiation-promotion assay. In 1997, the northamerican EPA evaluated Diuron as a “known/likely” carcinogen for humans (USEPA, 2004). In a previous study developed at this laboratory, male Wistar rats treated with Diuron 2500 ppm during 20 weeks presented increased indices of cell proliferation and incidences of simple urothelial hyperplasia (HS) in the urinary bladder. Under scanning electron microscopy (SEM) severe urothelial necrosis and hyperplasia were observed. However, in that study the urinary bladders of animals exposed to lower doses of Diuron were not examined under SEM. Therefore, the possible dose-response influence of Diuron on the urothelium under SEM is not known. The present study aimed to analyze under SEM the urinary bladder of male Wistar rats exposed to 125 ppm, 500 ppm and 2500 ppm doses of Diuron through diet during 20 weeks and to compare to the previous histological findings in the same material. Under SEM, 125 ppm and 2500 ppm groups presented significantly (p<0,05) increased incidences of simple hyperplasia, i.e., 7/10 and 8/10 respectively, compared to control group and the 500 ppm group The sensitivity of SEM was higher since it detected a 45% incidence of hyperplasiaswhile the histological analysis found only 27%. Considering SEM as the gold-standard, histology showed a 44% sensitivity, 86.4% specificity, a positive predictive value of 72,7% and negative predictive value of 65,5% and accuracy of 67,5%. Scanning Electron Microscopy...(Complete abstract click electronic access below)

Expressão do gene Glipican 3 (Gpc3) no urotélio da bexiga de ratos expostos ao diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea) por 7 dias e 20 semanas

Diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea) is a substituted urea herbicide widely used in crops of sugar cane, cotton and soybeans. In 1997, this agent has been classified by the United States Environmental Protection Agency as known/likely human carcinogen because it induced tumors in the urinary bladder and renal pelvis of rats, and breast and skin of mice exposed to 2500 ppm for feed for two years. A previous study from our group demonstrated dose-response relationship in the gene expression profile associated with severe necrosis on bladder urothelium and increased incidence of simple hyperplasia in male Wistar rats treated with different concentrations of diuron for 20 weeks. To check how early the molecular changes occurs, rats were fed for 7 days with diets containing diuron at 0, 125, 500 or 2500 ppm. The main observations recorded were urothelium ultrastructural alterations and disruptions of molecular pathways associated with cell-cell interaction and the tissue organization maintenance. Particularly, the gene Glypican 3 (Gpc3), a surface proteoglycan related to cellular adhesion and apoptosis induction, was down regulated on urothelium exposed to 2500ppm diuron for 7 days and 20 weeks. The aim of this study was validate by quantitative RT-PCR real time, the reduced Gpc3 gene expression in epithelial cells of the urinary bladder of male Wistar rats treated with different concentrations of diuron for 7 days and 20 weeks. The endogenous control of the quantitative PCR real time technique was the β-actin gene and the target was the gene Gpc3. The relative quantification (RQ) was obtained by the method of relative quantification 2-ΔΔCt . Animals exposed to diuron for 7 days or for 20 weeks presented reduction of Gpc3 gene expression compared to the control group. This reduction was statistically significant only for the 7 days study. Moreover, by comparing animals exposed for 7 days with the exposed for 20 weeks, it was ...

Principais aplicações terapêuticas da ozonioterapia

The ozone therapy is the therapeutic administration of ozone, which can be: subcutaneous (SC), intramuscular (IM) Intradiscal; intracavitary (pleural and peritoneal spaces); intravaginal, intrauretral, in the bladder; ozonated autohemotherapy. This therapy is being increasingly studied in order to help in some treatments and is being proven to be very effective in most cases, especially in acting on disinfection and healing of extensive wounds. There are over 6000 articles on the medical use of ozone in the literature, but the concentration used varies with each author. Most diseases have a positive response because ozone increases tissue oxygenation and metabolism. Discovered in Germany in the nineteenth century, ozone therapy still needs further study to clarify its mode of action and demonstrate its benefits. The objective of this review is to discuss some of the studies in the literature and try to clarify the main directions and forms of action of ozone therapy in medicine, showing the possibilities of getting good results including in veterinary medicine

Expansão ex vivo de células uroteliais em scaffolds 3D

Malformations and possible damages to the urogenital system can be originated in the embryonic period. Moreover, fire guns, knives and accidents, where there is the disruption of the urethra, also cause these lesions. The objective was to analyze the contribution of tissue engineering in the construction of neo-urethra, developed by bioengineering. We performed an urothelial ex vivo expansion of cells in 3D scaffolds (platelet gel matrix and acellular porcine aorta) to assess the contribution of this technique in the construction of a neo-urethra. Mechanical dissociation was made of the inner wall of 10 North Folk rabbit’s bladder, weighing 2.5 to 3.0 kg. After dissociation the cell content was centrifuged and obtained a pellet of urothelial cells. The pellet was ressuspended in culture medium DMEM F12 and cells were maintained in culture for 15 days. Immunohistochemical analysis characterized the urothelial culture. The cells were then implanted in the scaffold - platelet gel. In a second experiment using aortic porcine acellular matrix were implanted urothelial cells alone and urothelial cells on platelet gel, on the inner wall of the scaffold - aorta, with space for setting bordered by a urethral probe. The complex probe - cells - aorta and probe - cells in platelet gel - aorta, were sealed with suture material and culture were maintained in a humidified 37ºC incubator with 5% CO2 in air for 12 days to subsequent histological analysis of urothelium cell adhesion to the scaffolds. By observation under an optical microscope, we could see the growth of cells in the scaffold platelet gel, from a monolayer in to a three-dimensional structure. In the acellular porcine aortic matrix containing the platelet gel, we could observe a few quantity of urothelial cells adhered. However with the acellular porcine aortic matrix in which was implanted only the urothelial cells, we have obtained adhesion to the wall

Principais aspectos cirúrgicos da urolitíase em cães

Urolithiasis occurs with relatively frequency among the diseases that affect the urinary tract of dogs. It`s fundamental that samples of the uroliths are sent for analysis, because, in agreement with the mineral composition of the urolith and also its location, the choice is made between the surgical treatment or the clinical dissolution. Cystotomy is the surgical technique more accomplished in the bladder of small animals, due to its simplicity, when in comparison with other procedures for removal of calculi. With the aid of lithotripsy and endoscopy, the surgical interventions in the urinary treatment tend to be less and less invasive, minimizing the risks of subsequent complications. Chances of recurrence of the urolithiasis are usually larger in patients submitted to the surgical intervention. Besides, the time elapsed among regressive episodes is larger after the clinical dissolution

Female paraurethral prostate gland in Bitches

Even though the presence of the female prostate has been reported in many species, including humans, bats and several rodents, it has many anatomical and histological variations. There is still plentiful discussion on the biological function of this organ. Many authors state that paraurethral ducts and glands are functional and homologous to the male prostate. The use of experimental models and a better knowledge of the female prostate gland in other species, can be useful to veterinary medicine as well as human medicine. Therefore the aim of this study is to check for the presence of this gland in female dogs of various breeds and age. For that purpose 25 urethras, from the bladder to the vulva, were collected, fixed in 10% neutral buffered formalin, routinely processed and sectioned into 4 slides of 4 µm, each with 40 µm gap between each set of 4 slides, using an automatic microtome and stained with Hematoxylin and Eosine (HE). The HE sections were evaluated for the presence of prostatic gland in the sample. Unstained tissue sections cut from paraffin blocks were marked with a polyclonal anti-PSA primary antibody. The prevalence of the gland was 32% (8/25). The structure of the paraurethral PSA-positive gland was acinar, organized in buds, with secretory epithelium varying from cubic to columnar; eccentric nuclei, with lose chromatin and a layer of basal cells, very similar to the male prostate were observed. In view of these characteristics, for the first time in the literature, was demonstrate that those glands. may be considered as female prostate in dogs, as they are in other vertebrates

Dopplerfluxometria e avaliação morfométrica do fígado, baço, rins e grandes vasos abdominais de onças pardas (Puma concolor) da região Campo Grande, Mato Grosso do Sul, Brasil

Pós-graduação em Biotecnologia Animal - FMVZ

Avaliação dos efeitos de aloe-emodin em células uroteliais imortalizadas e neoplásicas cultivadas in vitro

Aloe vera (Aloe barbadensis Miller), popularly known in Brazil as babosa, has a long history of use as medicinal plant for different therapeutic purposes. The components of the plant extract are present in various products of human use, mainly for nutritional and cosmetics purposes. However, some studies suggest that this extract might also have carcinogenic activity. The aloe vera extract is a complex mixture of bioactive compounds. The study of isolated compounds may contribute to elucidate contradictory results about the effects related to the consumption of the plant, as well as their mechanisms of action. One of the most important compound from Aloe vera is aloe-emodin, which is a secondary metabolite generated in the intestinal tract. Putative antimicrobial and antitumor effects were previously attributed to aloe-emodin. Although the exposure of urothelial cells to aloe-emodin was already reported in the literature, only one study showed its effects on urothelial cells, suggesting that aloe-emodin inhibits the viability of T24 cancer cells due to apoptosis induction. Since there is no sufficient information about the effects of aloe-emodin on urothelial cells, and low efficiency in the treatment of bladder cancer currently, the present study aims to evaluate the hypothesis that the treatment with aloe-emodin could impact the behavior of other urothelial cell lines in vitro. Therefore, the in vitro IC50 exposure of aloe-emodin to human immortalized neoplastic urothelial cells will be determinated in order to verify possible differences in the behavior of urothelial cells in vitro treated with aloe-emodin in comparison with untreated cells. Furthermore, differences between cell lines will be also evaluated

Caracterização dos mecanismos de ação da imunoterapia intravesical com P-MAPA envolvendo as vias de sinalização dor receptores Toll-like (TLR) 2 e 4 na progressão do câncer de bexiga não-músculo invasivo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Induced reproduction and early development histology of Oscar Astronotus ocellatus (Agassiz, 1831)

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

Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors

Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients.

Bilateral hydronephrosis and hydroureter after ovariohysterectomy using nylon cable tie: a case report

A spayed crossbred female dog was presented due to progressive weight loss, emesis and anorexia over the preceding month. A complete blood count, urinalysis, serum biochemical panel, and ultrasound were initially performed. Computed tomography urography was performed as a complementary exam. Based on ultrasound and CT findings an exploratory celiotomy was performed to remove hyperdense structures that could be the cause of the hydronephrosis and hydroureter in both kidneys. An extensive granulomatous reaction was found near the caudal pole to the left kidney. A nylon cable tie adhering firmly to this tissue was removed during surgical excision. On the dorsal surface of the bladder an extensive granulomatous reaction that had entrapped the right ureter was also noted. Another nylon cable tie was removed and the ureter was released. Eight months postoperatively, the dog was in good general physical condition, showing appetite and vigour. Abdominal ultrasonography showed improvement of the hydronephrosis in both kidneys. The urea ratio was normal, but the creatinine level was slightly elevated, suggesting a guarded prognosis. Thus, bilateral hydronephrosis as observed in the present study should be considered as a major complication after elective ovariohysterectomy.

Effect of mesenchymal stem cells on movement and urination of rats with spinal cord injury

Cell therapy has frequently been reported as a possible treatment for spinal trauma in humans and animals; however, without pharmacologically curative action on damage from the primary lesion. In this study, we evaluated the effect of administering human adipose-derived stem cells (hADSC) in rats after spinal cord injury. The hADSC were used between the third and fifth passages and a proportion of cells were transduced for screening in vivo after transplantation. Spinal cord injury was induced with a Fogarty catheter no. 3 inserted into the epidural space with a cuff located at T8 and filled with 80 mu L saline for 5 min. The control group A (n = 12) received culture medium (50 mu L) and group B (n = 12) received hADSC (1.2 x 10(6)) at 7 and 14 days post-injury, in the tail vein. Emptying of the bladder by massage was performed daily for 3 months. Evaluation of functional motor activity was performed daily until 3 months post-injury using the Basso-Beattie-Bresnahan scale. Subsequently, the animals were euthanized and histological analysis of the urinary bladder and spinal cord was performed. Bioluminescence analysis revealed hADSC at the application site and lungs. There was improvement of urinary bladder function in 83.3% animals in group B and 16.66% animals in group A. The analysis of functional motor activity and histology of the spinal cord and urinary bladder demonstrated no significant difference between groups A and B. The results indicate that transplanted hADSC improved urinary function via a telecrine mechanism, namely action at a distance.

Alterações ultrassonográficas renais e vesicais de ovinos confinados e suplementados com cloreto de amônio

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

Mode B ultrasonography and abdominal Doppler in crab-eating-foxes (Cerdocyon thous)

Annually hundreds of crab-eating foxes (Cerdocyon thous) are referred to rehabilitation centers and zoos in Brazil. The ultrasonographic study of wildlife species is an important tool for a non-invasive and accurate anatomical description and provides important information for wildlife veterinary care. The aim of the present study was to determine the characteristics of the main abdominal organs as well as the vascular indexes of the abdominal aorta and renal arteries of crab-eating foxes (Cerdocyon thous) using mode B ultrasonography and Doppler ultrasonography, respectively. Ultrasonographic features of the main abdominal organs were described and slight differences were noticed between ultrasound imaging of abdominal organs of crab-eating foxes and other species. The bladder presented wall thickness of 12 +/- 0.01 mm, with three defined layers. Both, the right and left kidneys presented corticomedullary ratio of 1: 1 and similarly to the adrenals and the liver, they were homogeneous and hypoechoic compared to the spleen. The spleen was homogeneous and hyperechoic compared to the kidneys. The stomach presented 3 to 5 peristaltic movements per minute, wall thickness of 39 +/- 0.05 mm and lumen and mucosa with hyperechoic and hypoechoic features, respectively. Small and large intestines presented 2 to 3 peristaltic movements per minute, wall thickness of 34 +/- 0.03 mm and three defined layers with hyperechogenic (submucosa and serosa) and hypoechogenic (muscular) features. Ovaries of the female crab-eating fox were hypoechoic compared to the spleen and with heterogeneous parenchyma due to the presence of 2x2 mm ovarian follicles. Prostates of the six males were regular and with a well defined boundary, with a homogeneous and hyperechoic parenchyma compared to the spleen. Vascular indexes of the abdominal aorta (PSV: 25.60 +/- 0.32 cm/s; EDV: 6.96 +/- 1.68cm/s; PI: 1.15 +/- 0.07 e RI: 0.73 +/- 0.07) and right (PSV: 23.08 +/- 3.34cm/s; EDV: 9.33 +/- 2.36cm/s; PI: 1.01 +/- 0.65 e RI: 0.65 +/- 0.16) and left renal arteries (PSV: 23.74 +/- 3.94cm/s; EDV: 9.07 +/- 3.02cm/s; PI: 1.04 +/- 0.31 e RI: 0.64 +/- 0.10) were determined. Thus, conventional and Doppler ultrasonographic imaging provides basic information that can be used as reference for the species as well for other wild canids and it is a precise and non-invasive method that can be safely used to evaluate and diagnose abdominal injuries in these patients.