Estrutura, ultraestrutura e morfometria da veia cava de paca (Cuniculus paca Linnaeus, 1766) criada em cativeiro

A paca (Cuniculus paca) é o segundo maior roedor da fauna brasileira. Apresenta carne de excelente qualidade, o que incentiva a criação comercial. Além disso, este animal pode tornar-se uma opção válida em experimentação embora poucas sejam as informações detalhadas sobre sua morfologia. Assim, objetivou-se descrever a morfologia, morfometria e ultraestrutura de segmentos das porções cranial e caudal da veia cava de quatro pacas (Cuniculus paca) adultas excedentes do plantel do Setor de Animais Silvestres do Departamento de Zootecnia da FCAV-Unesp. Os segmentos venosos foram analisados à microscopia de luz e à microscopia eletrônica de varredura. Foram mensuradas as espessuras do complexo formado pelas túnicas íntima e média, além da túnica adventícia e analisou-se os resultados pela estatística descritiva, teste "T" pareado (p<0,05). Em relação à espessura das túnicas estudadas, comprovou-se que os valores da espessura das túnicas íntima, média e adventícia, para todos os animais, foram significativamente maiores no segmento cranial. As camadas das paredes dos vasos apresentaram variações entre si quanto à estrutura e espessura, supostamente devido a uma adaptação à exigência funcional.

An experimental model for the measurement of renal function after temporary ureteral obstruction in the rat

We present the results obtained with a ureterovesical implant after ipsilateral ureteral obstruction in the rat, suitable for the study of renal function after deobstruction in these animals. Thirty-seven male Wistar rats weighing 260 to 300 g were submitted to distal right ureteral ligation and divided into 3 groups, A (N = 13, 1 week of obstruction), B (N = 14, 2 weeks of obstruction) and C (N = 10, 3 weeks of obstruction). The animals were then submitted to ureterovesical implantation on the right side and nephrectomy on the left side. During the 4-week follow-up period serum levels of urea and creatinine were measured on the 2nd, 7th, 14th, 21st and 28th day and compared with preoperative levels. The ureterovesical implantation included a psoas hitch procedure and the ureter was pulled into the bladder using a transvesical suture. During the first week of the postoperative period 8 animals died, 4/13 in group A (1 week of obstruction) and 4/14 in group B (2 weeks of obstruction). When compared to preoperative serum levels, urea and creatinine showed a significant increase (P<0.05) on the 2nd postoperative day in groups A and B, with a gradual return to lower levels. However, the values in group B animals were higher than those in group A at the end of the follow-up. In group C, 2/10 animals (after 3 weeks of obstruction) were sacrificed at the time of ureterovesical implantation due to infection of the obstructed kidneys. The remaining animals in this group were operated upon but all of them died during the first week of follow-up due to renal failure. This technique of ureterovesical implantation in the rat provides effective drainage of the upper urinary tract, permitting the development of an experimental model for the study of long-term renal function after a period of ureteral obstruction

Signaled two-way avoidance learning using electrical stimulation of the inferior colliculus as negative reinforcement: effects of visual and auditory cues as warning stimuli

The inferior colliculus is a primary relay for the processing of auditory information in the brainstem. The inferior colliculus is also part of the so-called brain aversion system as animals learn to switch off the electrical stimulation of this structure. The purpose of the present study was to determine whether associative learning occurs between aversion induced by electrical stimulation of the inferior colliculus and visual and auditory warning stimuli. Rats implanted with electrodes into the central nucleus of the inferior colliculus were placed inside an open-field and thresholds for the escape response to electrical stimulation of the inferior colliculus were determined. The rats were then placed inside a shuttle-box and submitted to a two-way avoidance paradigm. Electrical stimulation of the inferior colliculus at the escape threshold (98.12 ± 6.15 (A, peak-to-peak) was used as negative reinforcement and light or tone as the warning stimulus. Each session consisted of 50 trials and was divided into two segments of 25 trials in order to determine the learning rate of the animals during the sessions. The rats learned to avoid the inferior colliculus stimulation when light was used as the warning stimulus (13.25 ± 0.60 s and 8.63 ± 0.93 s for latencies and 12.5 ± 2.04 and 19.62 ± 1.65 for frequencies in the first and second halves of the sessions, respectively, P<0.01 in both cases). No significant changes in latencies (14.75 ± 1.63 and 12.75 ± 1.44 s) or frequencies of responses (8.75 ± 1.20 and 11.25 ± 1.13) were seen when tone was used as the warning stimulus (P>0.05 in both cases). Taken together, the present results suggest that rats learn to avoid the inferior colliculus stimulation when light is used as the warning stimulus. However, this learning process does not occur when the neutral stimulus used is an acoustic one. Electrical stimulation of the inferior colliculus may disturb the signal transmission of the stimulus to be conditioned from the inferior colliculus to higher brain structures such as amygdala

Vasorelaxant effects of the potassium channel opener SR 47063 on the isolated human saphenous vein and rat aorta

The vasorelaxant effects of SR 47063 (4-(2-cyanimino-1,2-dihydropyrid-1-yl)-2,2-dimethyl-6-nitrochromene), a new K+-channel opener structurally related to levcromakalim, were examined in isolated human saphenous vein (HSV) and rat aorta (RA). HSV or RA rings were precontracted with either KCl or noradrenaline and cumulative relaxant concentration-response curves were obtained for SR 47063 (0.1 nM to 1 µM) in the presence or absence of 3 µM glibenclamide. SR 47063 potently relaxed HSV and RA precontracted with 20 mM (but not 60 mM) KCl or 10 µM noradrenaline in a concentration-dependent manner, showing slightly greater activity in the aorta. The potency of the effect of SR 47063 on HSV and RA was 12- and 58-fold greater, respectively, than that reported for the structurally related K+-channel opener levcromakalim. The vasorelaxant action of SR 47063 in both blood vessels was strongly inhibited by 3 µM glibenclamide, consistent with a mechanism of action involving ATP-dependent K+-channels.

Isolation and culture of umbilical vein mesenchymal stem cells

Bone marrow contains a population of stem cells that can support hematopoiesis and can differentiate into different cell lines including adipocytes, osteocytes, chondrocytes, myocytes, astrocytes, and tenocytes. These cells have been denoted mesenchymal stem cells. In the present study we isolated a cell population derived from the endothelium and subendothelium of the umbilical cord vein which possesses morphological, immunophenotypical and cell differentiation characteristics similar to those of mesenchymal stem cells isolated from bone marrow. The cells were isolated from three umbilical cords after treatment of the umbilical vein lumen with collagenase. The cell population isolated consisted of adherent cells with fibroblastoid morphology which, when properly stimulated, gave origin to adipocytes and osteocytes in culture. Immunophenotypically, this cell population was found to be positive for the CD29, CD13, CD44, CD49e, CD54, CD90 and HLA-class 1 markers and negative for CD45, CD14, glycophorin A, HLA-DR, CD51/61, CD106, and CD49d. The characteristics described are the same as those presented by bone marrow mesenchymal stem cells. Taken together, these findings indicate that the umbilical cord obtained from term deliveries is an important source of mesenchymal stem cells that could be used in cell therapy protocols.

Amino acid composition of parturient plasma, the intervillous space of the placenta and the umbilical vein of term newborn infants

The objective of the present study was to determine the levels of amino acids in maternal plasma, placental intervillous space and fetal umbilical vein in order to identify the similarities and differences in amino acid levels in these compartments of 15 term newborns from normal pregnancies and deliveries. All amino acids, except tryptophan, were present in at least 186% higher concentrations in the intervillous space than in maternal venous blood, with the difference being statistically significant. This result contradicted the initial hypothesis of the study that the plasma amino acid levels in the placental intervillous space should be similar to those of maternal plasma. When the maternal venous compartment was compared with the umbilical vein, we observed values 103% higher on the fetal side which is compatible with currently accepted mechanisms of active amino acid transport. Amino acid levels of the placental intervillous space were similar to the values of the umbilical vein except for proline, glycine and aspartic acid, whose levels were significantly higher than fetal umbilical vein levels (average 107% higher). The elevated levels of the intervillous space are compatible with syncytiotrophoblast activity, which maintain high concentrations of free amino acids inside syncytiotrophoblast cells, permitting asymmetric efflux or active transport from the trophoblast cells to the blood in the intervillous space. The plasma amino acid levels in the umbilical vein of term newborns probably may be used as a standard of local normality for clinical studies of amino acid profiles.

Efficacy of the FEV1/FEV6 ratio compared to the FEV1/FVC ratio for the diagnosis of airway obstruction in subjects aged 40 years or over

The present cross-sectional, population-based study was designed to evaluate the performance of the FEV1/FEV6 ratio for the detection of airway-obstructed subjects compared to the FEV1/FVC <0.70 fixed ratio test, as well as the lower limit of normality (LLN) for 1000 subjects ³40 years of age in the metropolitan area of São Paulo, SP, Brazil. After the exclusion of 37 (3.7%) spirometries, a total of 963 pre-bronchodilator (BD) and 918 post-BD curves were constructed. The majority of the post-BD curves (93.1%) were of very good quality and achieved grade A (762 curves) or B (93 curves). The FEV1/FEV6 and FEV1/FVC ratios were highly correlated (r² = 0.92, P < 0.000). Two receiver operator characteristic curves were constructed in order to express the imbalance between the sensitivity and specificity of the FEV1/FEV6 ratio compared to two FEV1/FVC cut-off points for airway obstruction: equal to 70 (area under the curve = 0.98, P < 0.0001) and the LLN (area under the curve = 0.97, P < 0.0001), in the post-BD curves. According to an FEV1/FVC <0.70, the cut-off point for the FEV1/FEV6 ratio with the highest sum for sensitivity and specificity was 0.75. The FEV1/FEV6 ratio can be considered to be a good alternative to the FEV1/FVC ratio for the diagnosis of airway obstruction, both using a fixed cut-off point or below the LLN as reference. The FEV1/FEV6 ratio has the additional advantage of being an easier maneuver for the subjects and for the lung function technicians, providing a higher reproducibility than traditional spirometry maneuvers.

Enhanced inhibition of murine prostatic carcinoma growth by immunization with or administration of viable human umbilical vein endothelial cells and CRM197

Vaccination with xenogeneic and syngeneic endothelial cells is effective for inhibiting tumor growth. Nontoxic diphtheria toxin (CRM197), as an immunogen or as a specific inhibitor of heparin-binding EGF-like growth factor, has shown promising antitumor activity. Therefore, immunization with or administration of viable human umbilical vein endothelial cells (HUVECs) combined with CRM197 could have an enhanced antitumor effect. Six-week-old C57BL/6J male mice were vaccinated with viable HUVECs, 1 x 10(6) viable HUVECs combined with 100 μg CRM197, or 100 μg CRM197 alone by ip injections once a week for 4 consecutive weeks. RM-1 cells (5 x 10(5)) were inoculated by sc injection as a preventive procedure. During the therapeutic procedure, 6-week-old male C57BL/6J mice were challenged with 1 x 10(5) RM-1 cells, then injected sc with 1 x 10(6) viable HUVECs, 1 x 10(6) viable HUVECs + 100 μg CRM197, and 100 μg CRM197 alone twice a week for 4 consecutive weeks. Tumor volume and life span were monitored. We also investigated the effects of immunization with HUVECs on the aortic arch wall and on wound healing. Vaccination with or administration of viable HUVECs+CRM197 enhanced the inhibition of RM-1 prostatic carcinoma by 24 and 29%, respectively, and prolonged the life span for 3 and 4 days, respectively, compared with those of only vaccination or administration with viable HUVECs of tumor-bearing C57BL/6J mice. Furthermore, HUVEC immunization caused some damage to the aortic arch wall but did not have remarkable effects on the rate of wound healing; the wounds healed in approximately 13 days. Treatment with CRM197 in combination with viable HUVECs resulted in a marked enhancement of the antitumor effect in the preventive or therapeutic treatment for prostatic carcinoma in vivo, suggesting a novel combination for anti-cancer therapy.

Dermatan sulfate reduces monocyte chemoattractant protein 1 and TGF-β production, as well as macrophage recruitment and myofibroblast accumulation in mice with unilateral ureteral obstruction

Selectins play an essential role in most inflammatory reactions, mediating the initial leukocyte-rolling event on activated endothelium. Heparin and dermatan sulfate (DS) bind and block P- and L-selectin function in vitro. Recently, we reported that subcutaneous administration of DS inhibits colon inflammation in rats by reducing macrophage and T-cell recruitment and macrophage activation. In the present study, we examined the effect of porcine intestinal mucosa DS on renal inflammation and fibrosis in mice after unilateral ureteral obstruction (UUO). Twenty-four adult male Swiss mice weighing 20-25 g were divided into 4 groups: group C (N = 6) was not subjected to any surgical manipulation; group SH (N = 6) was subjected to surgical manipulation but without ureter ligation; group UUO (N = 6) was subjected to unilateral ureteral obstruction and received no treatment; group UUO plus DS (N = 6) was subjected to UUO and received DS (4 mg/kg) subcutaneously daily for 14 days. An immunoblot study was also performed for TGF-β. Collagen (stained area ~3700 µm²), MCP-1 (stained area ~1700 µm²), TGF-β (stained area ~13% of total area), macrophage (number of cells ~40), and myofibroblast (stained area ~1900 µm²) levels were significantly (P < 0.05) higher in the UUO group compared to control. DS treatment significantly (P < 0.05) reduced the content of collagen (stained area ~700 µm²), MCP-1 (stained area ~160 µm²) and TGF-β (stained area ~5% of total area), in addition to myofibroblast (stained area ~190 µm²) and macrophage (number of cells ~32) accumulation in the obstructed kidney. Overall, these results indicate that DS attenuates kidney inflammation by reducing macrophage recruitment, myofibroblast population and fibrosis in mice submitted to UUO.

Disfunção do trato urinário inferior: um diagnóstico comum na prática pediátrica

Disfunção do trato urinário inferior indica uma função anormal do trato urinário inferior para a idade da criança, que pode levar à perda da capacidade coordenada de armazenamento e eliminação de urina. É uma entidade comum em crianças, embora subdiagnosticada na prática clínica, e que, além de representar um risco para o trato urinário superior, causa um constrangimento emocional aos pais e às crianças, devido à incontinência urinária e à frustração em lidar com o problema. A aquisição da continência urinária diurna ocorre na maioria das crianças até os 4 anos e a noturna até os 5 anos de idade. Após esta idade, a incontinência urinária torna-se um problema social. Apesar da importância clínica, muitas vezes, os pais desconhecem os sintomas dessa disfunção. Esse artigo tem como objetivo abordar os principais aspectos relacionados ao diagnóstico dessa disfunção.

Impacto de uma abordagem interdisciplinar em crianças e adolescentes com disfunção do trato urinário inferior (DTUI)

Introdução: A disfunção do trato urinário inferior (DTUI) corresponde a alterações no enchimento ou esvaziamento de urina de causas neurogênicas, anatômicas e funcionais. Objetivo: Avaliar o impacto do tratamento em crianças e adolescentes com DTUI. Métodos: Coorte histórica de 15 anos de seguimento com participação de 192 pacientes (123F, 69M) com idade inicial de 0,1 a 16,8 anos, analisados à admissão (T0) e ao final do seguimento (T1). A maioria dos pacientes era do grupo neurológico (60,4%). O tratamento instituído foi a uroterapia com intervenção comportamental e cognitiva, micção de hora marcada, hidratação oral, dieta laxativa, biofeedback, eletroestimulação sacral, cateterismo vesical intermitente limpo (CIL), terapia anticolinérgica, enema retal, tratamento da infecção do trato urinário (ITU) e, nos casos refratários, procedimentos cirúrgicos, tais como a derivação urinária continente e incontinente (vesicostomia), ampliação vesical e conduto para a realização do enema anterógrado cólico. Resultados: Os principais sintomas foram incontinência urinária diurna (82,3%), enurese noturna não monossintomática (78,6%), incontinência fecal (54,2%) e constipação intestinal (47,9%). Detectou-se redução significativa da infecção do trato urinário (p = 0,0027), da incontinência urinária diurna (p < 0,001), da enurese noturna (p < 0,001), da incontinência fecal (p = 0,010) e do refluxo vesicoureteral (p = 0,01). Houve aumento significativo no uso do CIL (p = 0,021), da terapia com anticolinérgico (p < 0,001) e diminuição da quimioprofilaxia (p < 0,001). Conclusão: Este estudo mostrou que o tratamento da DTUI na criança deve ser individualizado, além de requerer uma monitorização constante dos parâmetros clínicos, laboratoriais e de imagem, para minimizar o risco de lesão renal.

Redução da água corporal de renais crônicos em diálise peritoneal se associa com melhor controle da hipertensão arterial

Introdução: A hipertensão arterial tem alta prevalência em renais crônicos, sendo a hipervolemia um de seus fatores causais. Objetivo: Avaliar a influência da redução da volemia no controle pressórico e em parâmetros ecocardiográficos de pacientes renais crônicos em diálise peritoneal contínua. Métodos: Doze renais crônicos sem sinais clínicos de hipervolemia foram submetidos à intensificação da diálise com o objetivo de reduzir o peso corporal em 5%. A volemia foi avaliada pela bioimpedância elétrica e pela ultrassonografia de veia cava inferior (VCI). Os voluntários foram submetidos à monitorização ambulatorial da pressão arterial e a exame ecocardiográfico no período basal e após 5 semanas de intervenção. Resultados: Após a intensificação da ultrafiltração, houve redução significativa do peso corporal, da água extracelular e do diâmetro inspiratório da VCI, enquanto o índice de colapsamento da VCI não alterou de modo significativo. A despeito da redução do número de anti-hipertensivos, a pressão sistólica do período de sono reduziu de 138,4 ± 18,6 para 126,7 ± 18,0 mmHg, o descenso pressórico do sono aumentou e o diâmetro sistólico final do ventrículo esquerdo reduziu significantemente. Conclusão: A redução da volemia de pacientes em diálise peritoneal, clinicamente euvolêmicos, se associou a melhor controle pressórico e à diminuição do diâmetro sistólico final do ventrículo esquerdo.