Effects of Tributyltin (TBT) on Rat Bone and Mineral Metabolism.

BACKGROUND/AIMS: Tributyltin (TBT) is an organotin (OTs) and biohazard organometallic pollutant. Recently our group has shown that TBT, even in very low doses, has deleterious effects on several tissues most likely due to its role as an endocrine-disrupting molecule. Other studies have confirmed that OT exposure could be responsible for neural, endocrine, and reproductive dysfunctions via in vitro and in vivo models. However, TBT effects on bone lack concise data despite the fact that bone turnover is regulated by endocrine molecules, such as parathormone (PTH), estrogen (E2), etc. Our group has already shown that TBT disrupts adrenal and female gonadal functions. METHODS: We studied the effects of TBT on bone metabolism and structure using DXA, microCT scan, and SEM. We also determined the calcium (Ca²âº) and phosphate (Pi) metabolism in TBT-treated rats as well as some biomarkers for bone formation and resorption. RESULTS: Surprisingly, we found that TBT leads to higher bone mineral density (BMD) although lesions in spinal bone were observed by either microCT scan or SEM. Biomarkers for bone resorption, such as the urinary deoxipyridinolines (DPD) excretion ratio was increased in TBT-treated animals versus mock-treated controls. Osteocalcin (OC) and alkaline phosphatase (AP) are markers of bone formation and are also elevated suggesting that the bone matrix suffers from a higher turnover. Serum Ca²âº (total and ionized) do not changed by TBT treatment although hypercalciuria is observed. CONCLUSION: It is known that Sn atoms have three valence states (Sn²âº, Sn³âº, and Sn4⁺); hence, we hypothesized that Sn (more likely Sn²âº) could be competing with Ca²âº and/or Mg²âº in hydroxyapatite mineral matrix to disturb bone turnover. Further work is needed to confirm this hypothesis.

Videothermometry to evaluate metabolic activity in real time during pneumectomy in rats.

PURPOSE: To evaluate, in rats, the open field videothermometry in real time while performing left pneumonectomy for early diagnosis of cardiopulmonary changes. METHODS: Twelve non-specific pathogen-free Wistar rats were randomly allocated into two groups; pneumectomy group (GP) and sham surgery group (GS). Mean arterial pressure, videothermometry in real time, of the right lung, and histopathological analysis of the remaining lung were evaluated in all animals. RESULTS: Videothermometry in real time allowed identification of temperature variance of right lung after pneumectomy, indicating a significant decrease in temperature during evaluation. There was a statistical difference between M0 and M1, M1 and M2 and M0 and M2 (p<0.004) in GS, and significant difference between M0 and M1, M1 and M2, and M2 and M0 with p<0.0001 in GP. CONCLUSIONS: Left pneumonectomy in rats shows initial histopathological changes after 60 minutes of its completion, indicating a possible compensation beginning. The open-field videothermometry in real time proved to be efficient identifying the temperature changes of the remaining lung.

Videothermometry to evaluate metabolic activity in real time during pneumectomy in rats

Abstract Purpose: To evaluate, in rats, the open field videothermometry in real time while performing left pneumonectomy for early diagnosis of cardiopulmonary changes. Methods: Twelve non-specific pathogen-free Wistar rats were randomly allocated into two groups; pneumectomy group (GP) and sham surgery group (GS). Mean arterial pressure, videothermometry in real time, of the right lung, and histopathological analysis of the remaining lung were evaluated in all animals. Results: Videothermometry in real time allowed identification of temperature variance of right lung after pneumectomy, indicating a significant decrease in temperature during evaluation. There was a statistical difference between M0 and M1, M1 and M2 and M0 and M2 (p<0.004) in GS, and significant difference between M0 and M1, M1 and M2, and M2 and M0 with p<0.0001 in GP. Conclusions: Left pneumonectomy in rats shows initial histopathological changes after 60 minutes of its completion, indicating a possible compensation beginning. The open-field videothermometry in real time proved to be efficient identifying the temperature changes of the remaining lung.

Vitamin K Supplementation Modulates Bone Metabolism and Ultra-Structure of Ovariectomized Mice.

BACKGROUND/AIMS: Osteoporosis is a bone metabolic disease that affects mostly post-menopausal women. There has been shown that vitamin K (VK) supplementation during menopause may decrease bone loss as well as risk of bone breaking. Aiming to clarify the beneficial role of VK in bone metabolism during menopause, we investigated mineral metabolism and bone ultrastructure of ovariectomized (OVX) mice. METHODS: To determine the effects chronic use of VK in bone structure and mineral metabolism in OVX mice, we used several methods, such as DXA, µCTScan, and SEM as well as biomolecular techniques, such as ELISA and qRT-PCR. In addition, complete analysis of serum hormonal and other molecules associated to bone and lipid metabolism were evaluated overview the effects of VK in menopause murine model. RESULTS: VK treatment significantly affects Pi metabolism independently of OVX, changing Pi plasma, urinary output, balance, and Pi bone mass. Interestingly, VK also increased VLDL in mice independently of castration. In addition, VK increased compact bone mass in OVX mice when we evaluated it by DXA, histomorphometry, µCTScanning. VK increased bone formation markers, osteocalcin, HYP- osteocalcin, and AP whereas it decreased bone resorption markers, such as urinary DPD/creatinine ratio and plasmatic TRAP. Surprisingly, SEM images revealed that VK treatment led to amelioration of microfractures observed in OVX untreated controls. In addition, SHAM operated VK treated mice exhibited higher number of migrating osteoblasts and in situ secretion of AP. OVX led to decreased to in situ secretion of AP that was restored by VK treatment. Moreover, VK treatment increased mRNA expression of bone Calbindin 28KDa independently of OVX. CONCLUSION: VK treatment in OVX mice exhibited beneficial effects on bone ultrastructure, mostly by altering osteoblastic function and secretion of organic bone matrix. Therefore, VK could be useful to treat osteopenic/osteoporotic patients.

Vascular endothelial growth factor association with angiopoietin 1 promotes improvement in ventricular function after ischemic cardiomyopathy induced in mini pigs.

PURPOSE: To investigate the safety and clinical, hemodynamic and tissue improvement ability in mini pigs undergoing cell and gene therapy for the treatment of acute myocardial infarction. METHODS: Thirty-two mini pigs Br1 lineage, 12 months old, undergoing induction of acute myocardial infarction by occlusion of the diagonal branch of the paraconal coronary. They were divided into 4 groups: one control group and 3 treatment groups (cell therapy and gene cell therapy). Echocardiography reviews were performed on three occasions and histopathological analysis was performed after 4 weeks. Analysis of variance (ANOVA), Tukey and Wilcoxon tests, were performed. RESULTS: Association of vascular endothelial growth factor (VEGF) with angiopoietin1 (Ang1) presented satisfactory results in the improvement of ventricular function following ischemic cardiomyopathy in mini pigs when compared to the results of the other treated groups. CONCLUSION: The therapy with VEGF and the combination of VEGF with Ang1, promoted recovered function of the myocardium, characterized by reduced akinetic area and induction of neovascularization.

Vascular endothelial growth factor association with angiopoietin 1 promotes improvement in ventricular function after ischemic cardiomyopathy induced in mini pigs

Abstract Purpose: To investigate the safety and clinical, hemodynamic and tissue improvement ability in mini pigs undergoing cell and gene therapy for the treatment of acute myocardial infarction. Methods: Thirty-two mini pigs Br1 lineage, 12 months old, undergoing induction of acute myocardial infarction by occlusion of the diagonal branch of the paraconal coronary. They were divided into 4 groups: one control group and 3 treatment groups (cell therapy and gene cell therapy). Echocardiography reviews were performed on three occasions and histopathological analysis was performed after 4 weeks. Analysis of variance (ANOVA), Tukey and Wilcoxon tests, were performed. Results: Association of vascular endothelial growth factor (VEGF) with angiopoietin1 (Ang1) presented satisfactory results in the improvement of ventricular function following ischemic cardiomyopathy in mini pigs when compared to the results of the other treated groups. Conclusion: The therapy with VEGF and the combination of VEGF with Ang1, promoted recovered function of the myocardium, characterized by reduced akinetic area and induction of neovascularization.

Technical innovation: Intragastric Single Port Sleeve Gastrectomy (IGSG). A feasibility survival study on porcine model.

PURPOSE: To perform technically the laparoscopic sleeve gastrectomy (LSG) using a unique Intragastric Single Port (IGSG) in animal swine model, evidencing an effective and safe procedure, optimizing the conventional technique. METHODS: IGSG was performed in 4 minipigs, using a percutaneous intragastric single port located in the pre-pyloric region. The gastric stapling of the greater curvature started from the pre-pyloric region towards the angle of His by Endo GIA™ system and the specimen was removed through the single port. In the postoperative day 30, the animals were sacrificed and submitted to autopsy. RESULTS: All procedures were performed without conversion, and all survived 30 days. The mean operative time was 42 min. During the perioperative period no complications were observed during invagination and stapling. No postoperative complications occurred. Post-mortem examination showed no leaks or infectious complications. CONCLUSION: Intragastric Single Port is a feasible procedure that may be a suitable alternative technique of sleeve gastrectomy for the treatment of morbid obesity.

Technical innovation: Intragastric Single Port Sleeve Gastrectomy (IGSG). A feasibility survival study on porcine model

Abstract Purpose: To perform technically the laparoscopic sleeve gastrectomy (LSG) using a unique Intragastric Single Port (IGSG) in animal swine model, evidencing an effective and safe procedure, optimizing the conventional technique. Methods: IGSG was performed in 4 minipigs, using a percutaneous intragastric single port located in the pre-pyloric region. The gastric stapling of the greater curvature started from the pre-pyloric region towards the angle of His by Endo GIA™ system and the specimen was removed through the single port. In the postoperative day 30, the animals were sacrificed and submitted to autopsy. Results: All procedures were performed without conversion, and all survived 30 days. The mean operative time was 42 min. During the perioperative period no complications were observed during invagination and stapling. No postoperative complications occurred. Post-mortem examination showed no leaks or infectious complications. Conclusion: Intragastric Single Port is a feasible procedure that may be a suitable alternative technique of sleeve gastrectomy for the treatment of morbid obesity.

Subcolectomy and symphyseal distraction-osteotomy using a spacer of spirally fashioned orthopedic wire: a treatment option for cats with pelvic canal stenosis, megacolon and obstipation / Subcolectomia e distração da sínfise púbica com espaçador de fio ortopédico espiralado: uma opção de tratamento para gatos com estenose do canal pélvico, megacolon e obstipação

ABSTRACT: The use of two cerclages with wire spirals as distractor spacers was evaluated for the correction of pelvic stenosis in felines. A three-year-old, mixed-breed, male feline was taken to the Veterinary Hospital of the UFRJ. It presented symptoms of constipation, dyschezia, hyporexia and mild apathy. On radiographic examination, a deformation of the pelvic bones and a narrowing of the pelvic canal were detected. The patient underwent pelvic stenosis correction surgery. Two spirals were made with cerclage wire and 2.0-mm diameter Steinmann pins; coupled to the chuck of a drill, the pins were wrapped with cerclage wire, running laps around the pin. Then, the coils were positioned in the space formed by separating the hemipelves and stabilized by keeping them separated to the desired opening. Effectiveness of the technique was evidenced by measurements made using new radiographic images in comparison with the images obtained before the surgical procedure. There were evident decreases in the sacro-ischium line and pubic line of 3.98 and 3.18, demonstrating an increase of approximately 21% in the pelvic canal diameter. The animal showed no apparent macroscopic or radiographic reaction. The preliminary clinical application of the adapted pelvic distraction technique is relevant for allowing the appropriate correction of a stenosis of the pelvic girdle and its consequences, representing a major technical and scientific innovation in the correction.

RESUMO: Avaliou-se o uso de duas espirais confeccionadas com fio de cerclagem como preenchimento distrator para correção de estenose pélvica em felinos. Foi atendida no Hospital Veterinário da UFRJ um felino macho, sem raça definida com idade de 3 anos, apresentando sintomas de constipação, disquezia, hiporexia e apatia leve. Ao exame radiográfico, observou-se deformação de ossos da pelve com estreitamento do canal pélvico. O animal foi operado para correção da estenose pélvica. Foram confeccionados dois espirais com fio de cerclagem e um pino de Steinmann de 2,0mm de diâmetro acoplado ao mandril de uma perfuratriz, envolveu-se este pino com o fio de cerclagem, executando voltas ao redor do pino. Em seguida, os espirais foram posicionados no espaço formado pela separação das hemipelves, e estabilizados, mantendo-as separadas com a abertura desejada. A efetividade da técnica foi evidenciada por medições feitas através de novas imagens radiográficas, permitindo assim uma comparação com a imagem obtida anteriormente ao procedimento cirúrgico. A aplicação clínica da técnica é relevante por permitir que a correção apropriada da estenose da cintura pélvica e suas consequências representa uma importante inovação técnica e científica na correção dessa condição.

Ligation of the mesovarium in dogs with a self-locking implant of a resorbable polyglycolic based co-polymer: a study of feasibility and comparison to suture ligation.

BACKGROUND: Ligation of the mesovarium in female dogs may be cumbersome with risk of complications and is associated with intense noxious stimuli. A resorbable implant, a self-locking loop designed for surgery, was developed as an alternative to traditional ligation. The study aimed to test the feasibility of ligating the canine ovarian pedicle with the implant and to compare its performance to traditional suture ligation. RESULTS: In total 45 intact female dogs destined for elective ovariohysterectomy and adoption were included. In 21 dogs the new resorbable implant was used to ligate the mesovarium, and in 24 control dogs traditional suture was used with one encircling ligature. Mean weight of implant dogs was 10.7 ± 5.6 kg (range 3.5-22.0), and mean weight of control dogs was 12.8 ± 6.4 kg, (range 4.1-27.0). The body weight of dogs did not differ between groups (P = 0.25). In total, 42 ovarian pedicles were successfully ligated with the implant. In one control dog, intraoperative haemorrhage from the left ovarian pedicle was diagnosed. The mesovarium was re-ligated and haemostasis was confirmed. All dogs recovered uneventfully. The ligation time of the mesovarium was significantly shortened (P = 0.02) by using the self-locking implant versus a single ligature (3'28'' ± 1'05'' and 5'29'' ± 3'54'', respectively). Total duration of surgery differed between the groups (P = 0.02) with a shortened duration of surgery when using the self-locking implant (15'56'' ± 2'47'' and 20'39'' ± 8'58'', study group versus control group, respectively). In both groups, duration of surgery and time required to ligate the ovarian pedicle were longer in larger dogs than smaller dogs. CONCLUSION: The results of this feasibility study suggested the implant can be used to ligate the canine mesovarium. Compared with traditional suture ligation, the results suggested that time to ligate the ovarian pedicle and duration of surgery were significantly reduced with the implant. More time was required to perform surgeries in larger dogs.

Oxigenação extracorpórea por membrana em cães sem auxílio circulatório / Pumpless extracorporeal membrane oxygenation in dogs

A oxigenação extracorpórea por membrana com desvio arterio-venoso (AV-ECMO), em seres humanos, é uma técnica de suporte à vida capaz de assumir em parte ou totalmente as funções dos pulmões, quando estes já não são capazes de promover uma adequada hematose. Para conhecer a viabilidade deste método em cães (Canis familiaris), o presente estudo aplicou a AV-ECMO em dez indivíduos adultos, sem raça definida, com uma expectativa de três horas de duração para cada experimento. No grupo 1 (n=4), ventilação mecânica e infusão contínua de dopamina foram associadas ao suporte extracorpóreo. No grupo 2 (n=6), nenhuma medida de terapia intensiva foi instituída concomitantemente à AV-ECMO. O comportamento dos parâmetros de gasometria e os níveis de lactato foram avaliados a cada 10 minutos e 1 hora, respectivamente, por meio de da avaliação de amostras sanguíneas arteriais. O suporte inotrópico foi capaz de promover uma melhor perfusão tecidual com sangue oxigenado, sugerida pelos níveis de lactato e pressão arterial média mensurados, assim como a hemogasometria revelou melhores resultados quando a ventilação mecânica foi utilizada simultaneamente à AV-ECMO. Portanto, aplicada conjuntamente com suporte da terapia intensiva, a AV-ECMO é uma técnica viável para o uso em cães com falência respiratória, refratária aos tratamentos convencionais.

In humans, arteriovenous extracorporeal membrane oxygenation (AV-ECMO) is a life support technique capable to assume in part or totally lungs functions, when these are not able to promote appropriated hematosis. To know the viability of this method in dogs (Canis familiaris), the present study applied AV-ECMO in ten adult mixed-breed dogs, with expected three hours of duration to each experiment. In Group 1 (n=4), mechanical ventilation and dopamine continuous infusion were associated with the extracorporeal support. In Group 2 (n=6), any other measure of intensive care was used concomitantly AV-ECMO. Blood gas analysis and lactate levels were evaluated every 10 minutes and one hour, respectively, from arterial blood samples. Inotropic support improved the blood flow through the circuit promoting a better tissue perfusion with oxygenated blood, suggested by the lactate and mean arterial pressure levels measured. Mechanical ventilation used simultaneously with AV-ECMO was able to further improve levels of blood gases. Therefore, if applied along intensive care support, AV-ECMO is a viable technique for use in dogs with respiratory failure refractory to conventional treatments.

Osteotomia de nivelamento do platô tibial no tratamento da ruptura do ligamento cruzado cranial estudo clínico em cães / Tibial plateau leveling osteotomy on the treatment cranial cruciate ligament rupture – clinic study in dogs

A ruptura do ligamento cruzado cranial (RLCC) é a causa comum de claudicação no membro pélvico em cães. Estudosrecentes demonstram que o ângulo de inclinação do platô tibial está associado à RLCC. A partir dessa descoberta,desenvolveu-se a técnica de osteotomia e nivelamento do platô tibial (TPLO). A técnica consiste na osteotomia, rotação eestabilização da porção proximal da tíbia, alterando a mecânica da articulação, neutralizando o impulso tibial cranial. Aproposta do presente estudo é revisar a técnica de TPLO, enfatizando o procedimento, técnica cirúrgica, cuidados pósoperatóriose complicações. O grupo estudado era composto de 40 cães. Dezessete pacientes apresentaram RLCC nomembro pélvico esquerdo e 23 no membro pélvico direito. Os ângulos do platô tibial variaram de 16o 36o. Trinta animais(75%) voltaram a apoiar o membro operado nos primeiros sete dias, enquanto seis animais (15%) apoiaram até o 15o dia,um animal (2,5%) apoiou até o 21o dia e outro (2,5%) até o 28o dia. Três animais (7,5%) apresentaram complicações pósoperatórias,sendo elas: deiscência de sutura, fratura tibial, falha no implante. A técnica de TPLO mostrou-se eficaz notratamento de cães com diferentes pesos com RLCC. O apoio precoce do membro afetado ocorreu na maioria dos animais.As complicações encontradas, que envolviam o procedimento cirúrgico, puderam ser corrigidas.

Cranial cruciate ligament rupture (CCLR) frequently causes hindlimb limping in dogs. Recent studies showed that tibial plateau angle shift is associated with CCLR. Based on this founding the tibial plateau leveling osteotomy (TPLO) technique was developed. It consists about osteotomy, rotation and stabilization of tibial proximal portion, which modify joint mechanics thus neutralizing tibial cranial thrust. The proposal of this study is to report a review of the TPLO procedure, emphasizing procedure, surgical technique, post operative care and complications. Fourty dogs presenting CCLR were submitted to surgery, seventeen with the left limb affected, 23 on the right. The tibial plateau angles before surgery varied from 16o to 36o. Thirty animals (75%) returned to limb normal use in a week; six animals (15%) were back to normal deambulation to the fifteenth day, one animal (2,5%) were back to normal deambulation to the 200 day other one before the 30o day. Three animals (7,5%) had surgical post-operatory complications, including suture descending, tibial fracture and implant failure. TPLO technique seemed appropriate for treatment of CCLR from dogs within a broad weight range. Early limb use occurred in most animals. Eventual problems arose from surgical procedure, and were promptly corrected.

Estudo comparativo entre as técnicas de sutura total e serosubmucosa em anastomoses colônicas na presença de um protetor intraluminar em cães / Comparative study of all-layers suture and extramucosal single layer suture with an intraluminal protector in dogs

OBJETIVO: Avaliar a cicatrização comparando as técnicas de sutura total e parcial em anastomoses colônicas término-terminais na presença de um protetor intraluminar confeccionado a partir de um dreno de Penrose. MÉTODOS: Foram realizadas anastomoses colônicas em 10 cães nos quais foi suturado a mucosa / submucosa um protetor intraluminar por meio da eversão do segmento intestinal proximal em 10 cm. Os animais foram distribuidos em dois grupos experimentais: Grupo 1 - sutura total e Grupo 2 - sutura serosubmucosa. Os animais foram reoperados com sete dias de pós-operatório para coleta de material e subseqüente análise histopatológica. RESULTADOS: Observamos no exame histopatológico das anastomoses realizadas com sutura serosubmucosa (Grupo 2) uma melhor cicatrização caracterizada por um menor infiltrado inflamatório e por uma maior integridade das camadas intestinais quando comparadas com o Grupo 1. CONCLUSÃO: O uso da técnica de sutura parcial mostrou-se superior a de sutura total nas anastomoses colônicas na presença de protetor intraluminar.

BACKGROUND: This study aims to evaluate the colonic anastomotic healing testing of all layers suture and single layer suture using an intraluminal protector. The intraluminal protector was shaped from a Penrose drain and then fixed 10 cm from the superior end of the anastomotic site. METHODS: Colonic anastomoses were done in 10 dogs, which were sutured, in the proximal segment with an intraluminal protector made of Penrose n.3 drain, using interrupted mucosal / submucosal suture. In the subsequent anastomoses, the animals were subdivided in two experimental groups: 1 - in all layers and 2 - extramucosal. In the seventh postoperative day, a tissue fragment was collected for histopathological analysis. RESULTS: Group 2 histopathological examinations had a better scar (extramucosal suture), characterized by a lesser inflammatory process than in animals in which all layers suture was used (group 1). CONCLUSION: The use of extramucosal suture was better than total suture in colonic anastomoses with the presence of an intraluminal protector.

Use of intraluminal protection in colonic anastomosis in dogs.

PURPOSE: To test the use of intraluminal protection in colonic anastomosis without intestinal cleansing. The intraluminal liner was fashioned from porcine submucosa preserved in glycerin and then fixed 10 cm anteriorly to the anastomotic site. This technique was compared with the one used in termino-terminal colonic anastomosis without intraluminal protection. METHODS: Twenty-eight dogs were divided into two groups of fourteen animals each. Clinical and histopathological tests were performed on the fourth and twenty-first postoperative days. RESULTS: The morbidity and mortality rates were higher in animals that did not receive the intraluminal liner. Histopathological examinations in animals in which the intraluminal liner was used showed better healing, characterized by milder inflammation and increased amount of collagen. CONCLUSION: It can be concluded that the use of intraluminal protection decreases complication rates in colonic anastomosis and promotes better healing.

Use of intraluminal protection in colonic anastomosis in dogs

PURPOSE: To test the use of intraluminal protection in colonic anastomosis without intestinal cleansing. The intraluminal liner was fashioned from porcine submucosa preserved in glycerin and then fixed 10 cm anteriorly to the anastomotic site. This technique was compared with the one used in termino-terminal colonic anastomosis without intraluminal protection. METHODS: Twenty-eight dogs were divided into two groups of fourteen animals each. Clinical and histopathological tests were performed on the fourth and twenty-first postoperative days. RESULTS: The morbidity and mortality rates were higher in animals that did not receive the intraluminal liner. Histopathological examinations in animals in which the intraluminal liner was used showed better healing, characterized by milder inflammation and increased amount of collagen. CONCLUSION: It can be concluded that the use of intraluminal protection decreases complication rates in colonic anastomosis and promotes better healing.

OBJETIVO: Testar o uso da proteção intraluminal na anastomose colônica sem preparo intestinal. O protetor intraluminal usado foi confeccionado a partir da submucosa de suíno conservada em glicerina, e fixado a 10 cm cranialmente ao sítio anastomótico. Essa técnica foi comparada com a técnica de anastomose colônica término-terminal sem uso do protetor intraluminal. MÉTODOS: Foram utilizados 28 cães divididos em dois grupos de 14 animais cada. A avaliação foi através de exames clínicos e histopatológicos. A avaliação anatomo-patológica foi realizada no quarto e vigésimo primeiro dias de pós-operatório. RESULTADOS: Um maior número de casos de morbi-mortalidade foi observado nos animais operados sem o protetor intraluminal. O exame histopatológico dos animais nos quais foram usados os protetores intraluminais mostrou melhor cicatrização, caracterizada por processo inflamatório mais discreto e maior quantidade de colágeno. CONCLUSÃO: O uso do protetor diminui o número de complicações em anastomoses de cólon e melhora a cicatrização.