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.

Diuretic and hipotensive activity of aqueous extract of parsley seeds (Petroselinum sativum Hoffm.) in rats

A espécie vegetal, Petroselinum sativum Hoff, conhecida como salsa, é amplamente utilizada na medicina popular brasileira como diurético. O objetivo desse estudo é verificar se o uso brasileiro do extrato aquoso da salsa tem efeitos semelhantes com investigações que mostram o efeito diurético da P. sativum em ratos. MÉTODOS: 19 Ratos foram anestesiados e canulamos a traquéia, artéria carótida esquerda (para a medição da pressão arterial) e bexiga urinária (para coletar urina). Depois de 40 minutos para adaptação das condições cirúrgicas, ratos anestesiados foram administrados de acordo com seus grupos: controle (CON), administração oral com 1.0 mL de água filtrada, e grupo tratado (AE), administração oral com extrato aquoso de sementes de salsa 20% (AE). Urina foi coletada três vezes (de 30 em 30 minutos) e então esse material foi utilizado para determinações de sódio e potássio, para avaliar a quantidade excretada desses íons. Pressão arterial foi medida pelo manômetro de mercúrio por 9 vezes. Todos os dados foram estatisticamente avaliados. RESULTADOS E CONCLUSÃO: nos parâmetros anestesiados, o grupo CON não mostrou nenhuma diferença; mas o grupo AE mostrou um aumento do fluxo urinário e da quantidade excretada de sódio e potássio, e também uma diminuição da pressão arterial. Todos os parâmetros apresentaram essas modificações após 30 minutos de administração do AE (p<0,05). Esses resultados mostram que o tratamento com o AE leva a efeitos natriurético e hipotensor em ratos Wistar anestesiados, confirmando o uso da população brasileira dessa erva como diurético.

Mirabegron relaxes urethral smooth muscle by a dual mechanism involving β3-adrenoceptor activation and α1-adrenoceptor blockade

Mirabegron is the first β3-adrenoceptor (AR) agonist approved for treatment of overactive bladder syndrome (OAB). This study aimed to investigate the effects of β3-adrenoceptor (AR) agonist mirabegron in mouse urethra. The possibility that mirabegron exerts α1-AR antagonism was also tested in rat smooth muscle preparations presenting α1A- (vas deferens and prostate), α1D- (aorta) and α1B-AR (spleen). Functional assays were carried out in mouse and rat isolated tissues. Competition assays for the specific binding of [(3) H]Prazosin to membrane preparations of HEK 293 cells expressing each of the human α1-ARs, as well as β-AR mRNA expression and cyclic AMP measurements in mouse urethra were performed. Mirabegron produced concentration-dependent urethral relaxations that were right shifted by the selective β3-AR antagonist L 748,337, but unaffected by β1- and β2-AR antagonists (atenolol and ICI 118,551, respectively). Mirabegron-induced relaxations were enhanced by the phosphodiesterase-4 inhibitor rolipram, and this agonist stimulated cAMP synthesis. Mirabegron also produced rightward shifts in urethral contractions induced by the α1-AR agonist phenylephrine. Schild regression analysis revealed that mirabegron behaves as a competitive antagonist of α1-AR in urethra, vas deferens and prostate (α1A-AR, pA2  ≅ 5.6) and aorta (α1D-AR, pA2  ≅ 5.4), but not in spleen (α1B-AR). The affinities estimated for mirabegron in functional assays were consistent with those estimated in radioligand binding with human recombinant α1A- and α1D-ARs (pKi ≅ 6.0). The effects of mirabegron in urethral smooth muscle are the result of β3-AR agonism together with α1A / α1D-AR antagonism.

Uncaria tomentosa increases growth and immune activity in Oreochromis niloticus challenged with Streptococcus agalactiae

Cat's claw (Uncaria tomentosa) is an Amazon herb using in native cultures in Peru. In mammals, it has been described several effects of this herb. However, this is the first report of its use on the diet of fish. The aim of this study was to determinate the effect of this plant on the growth and immune activity in Oreochromis niloticus. Nile tilapia (81.3 ± 4.5 g) were distributed into 5 groups and supplemented with 0 (non-supplement fish), 75, 150, 300, and 450 mg of U. tomentosa.kg(-1) of diet for a period of 28 days. Fish were inoculated in the swim bladder with inactivated Streptococcus agalactiae and samples were taken at 6, 24, and 48 h post inoculation (HPI). Dose dependent increases were noted in some of the evaluated times of thrombocytes and white blood cells counts (WBC) in blood and exudate, burst respiratory activity, lysozyme activity, melanomacrophage centers count (MMCs), villi length, IgM by immunohistochemistry in splenic tissue, and unexpectedly on growth parameters. However, dietary supplementation of this herb did not affect red blood cells count (RBC), hemoglobin, and there were no observed histological lesions in gills, intestine, spleen, and liver. The current results demonstrate for the first time that U. tomentosa can stimulate fish immunity and improve growth performance in Nile tilapia.

Cytotoxic and genotoxic effects of two hair dyes used in the formulation of black color

According to the International Agency for Research on Cancer (IARC), some hair dyes are considered mutagenic and carcinogenic in in vitro assays and exposed human populations. Epidemiological studies indicate that hairdressers occupationally exposed to hair dyes have a higher risk of developing bladder cancer. In Brazil, 26% of the adults use hair dye. In this study, we investigated the toxic effects of two hair dyes, Basic Red 51 (BR51) and Basic Brown 17 (BB17), which are temporary dyes of the azo group (R-N=N-R'), used in the composition of the black hair dye. To this end, MTT and trypan blue assays (cytotoxicity), comet and micronucleus assay (genotoxicity) were applied, with HepG2 cells. For cytotoxic assessment, dyes were tested in serial dilutions, being the highest concentrations those used in the commercial formula for hair dyes. For genotoxic assessment concentrations were selected according to cell viability. Results showed that both dyes induced significant cytotoxic and genotoxic effects in the cells, in concentrations much lower than those used in the commercial formula. Genotoxic effects could be related to the azo structure present in the composition of the dyes, which is known as mutagenic and carcinogenic. These results point to the hazard of the hair dye exposure to human health.

Efeitos das formulações nanoestruturadas de doxorrubicina e cisplatina em dispersão de óxido de grafeno reduzido no tratamento da progressão do câncer de bexiga Não-músculo invasivo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Eficácia e efeitos hemodinâmicos da anestesia raquidiana com ropivacaína isobárica, hipobárica ou hiperbárica seletiva em cães anestesiados com isofluorano

Pós-graduação em Ciência Animal - FMVA

Avaliação dos dados obtidos em um programa de esterilização cirúrgica de gatos em Araçatuba, SP no período de 1996 a 2010

Cats are gradually occupying a more important position as pets and this preference is a result of how easy cat maintenance in an urban environment is, even though they are very prolific and need surgical sterilization. This paper aims at evaluating obtained data within 15 years of research in a university service program that offers free cats sterilization surgery. We carried out a statistical analysis of data shown in the records of animals treated by the program. Surgical sterilization was performed on 647 animals (409 females – 63% and 238 males – 37%). Unilateral cryptorchidism was observed in 9 (3.8%) male cats. Forty (10%) female cats were pregnant at the time of the surgery and the treatment with contraceptives was observed in 67 (16.4%) female cats. One death occurred during anesthesia recovery and 2 cats were euthanized because of wound infection, totalizing an amount of 0.5% of severe complications. From 1996 to 2004, 212 female cats were spayed (122 adults and 90 prepubertal) and during that time two different approaches for ovariohysterectomy were compared: flank laparotomy and ventral midline celiotomy. The flank laparotomy approach was used in only 46 female cats (21.7%) due to some disadvantages observed – the need of an incision on each flank in prepubertal or nulliparous animals and the difficulty or impossibility of total uterus removal. In the same period, 105 male cats underwent orchiectomy via an open technique in which the spermatic cord was linked with nylon thread. From 2005 to 2010, 197 females (106 adults and 91 prepubertal) were spayed. The minilaparotomy technique was used to perform ovariohysterectomy on 139 female cats (70.6%). In this procedure, ovaries and uterus were exteriorized in a blind fashion with a hook through a small midline incision. The traditional midline ovariohysterectomy, which incision length permits direct visualization of the ovaries and uterus, had to be performed in 58 (29.4%) female cats due to advanced pregnancy, full urinary bladder during surgery or obesity. Over the past 6 years, 133 male cats (48 adults and 85 prepubertal) were castrated by means of an open technique in which the spermatic cord was tied to itself. The demand for surgeries during the project demonstrated that people are becoming aware concerning the importance of sterilization when facing cat overpopulation and abandonment.

Atividade toxicogenética de corantes usados na formulação da tintura capilar preta, individuais e em associação, por meio de diferentes ensaios biológicos

Pós-graduação em Ciências Biológicas (Biologia Celular e Molecular) - IBRC