Ney Luis Pippi

Anthocyanins suppress the secretion of proinflammatory mediators and oxidative stress, and restore ion pump activities in demyelination

The aim of this study was to investigate the protective effect of anthocyanins (ANT) on oxidative and inflammatory parameters, as well as ion pump activities, in the pons of rats experimentally demyelinated with ethidium bromide (EB). Rats were divided in six groups: control, ANT 30 mg/kg, ANT 100 mg/kg, EB (0.1%), EB plus ANT 30 mg/kg and EB plus ANT 100 mg/kg. The EB cistern pons injection occurred on the first day. On day 7, there was a peak in the demyelination. During the 7 days, the animals were treated once per day with vehicle or ANT. It was observed that demyelination reduced Na(+),K(+)-ATPase and Ca(2+)-ATPase activities and increased 4-hydroxynonenal, malondialdehyde, protein carbonyl and NO2plus NO3 levels. In addition, a depletion of glutathione reduced level/nonprotein thiol content and a decrease in superoxide dismutase activity were also seen. The dose of 100 mg/kg showed a better dose-response to the protective effects. The demyelination did not affect the neuronal viability but did increase the inflammatory infiltrate (myeloperoxidase activity) followed by an elevation in interleukin (IL)-1β, IL-6, tumor necrosis factor-α and interferon-γ levels. ANT promoted a reduction in cellular infiltration and proinflammatory mediators. Furthermore, ANT restored the levels of IL-10. Luxol fast blue staining confirmed the loss of myelin in the EB group and the protective effect of ANT 100 mg/kg. In conclusion, this study was the first to show that ANT are able to restore ion pump activities and protect cellular components against the inflammatory and oxidative damages induced by demyelination.

Renal and cardiorespiratory effects of treatment with lactated Ringer's solution or physiologic saline (0.9% NaCl) solution in cats with experimentally induced urethral obstruction.

OBJECTIVE To compare the renal and cardiorespiratory effects of IV treatment with lactated Ringers solution (LRS) or physiologic saline (0.9% NaCl) solution (PSS) in severely decompensated cats with urethral obstruction (UO). ANIMALS 14 cats (4 cats were used only to establish infusion rates). PROCEDURES An occluded urethral catheter was used to induce UO in each cat. After development of severe metabolic acidosis, hyperkalemia, and postrenal azotemia, the obstruction was relieved (0 hours); LRS or PSS (5 cats/group) was administered IV (gradually decreasing rate) beginning 15 minutes before and continuing for 48 hours after UO relief. Ten minutes before urethral catheter placement (baseline), at start of fluid therapy (SFT), and at intervals during fluid administration, various physical and clinicopathologic evaluations were performed. RESULTS Metabolic acidosis was detected in the PSS-treated group at SFT and 2 hours after relief of UO and in the LRS-treated group only at SFT The PSS-treated group had significantly lower blood pH and bicarbonate concentrations at 8 through 48 hours and lower base excess values at 2 through 48 hours, compared with the LRS-treated group. Hypocalcemia and hypernatremia were detected in the PSS-treated group at 2 and 12 hours, respectively. Absolute serum potassium and chloride concentrations did not differ significantly between groups at any time point. CONCLUSIONS AND CLINICAL RELEVANCE Treatment with LRS or PSS appeared to be safe and effective in cats with experimentally induced UO; however, LRS was more efficient in restoring the acid-base and electrolyte balance in severely decompensated cats with UO.

Laparoscopia nas hérnias diafragmáticas: estudo experimental em cães

The most common type of diaphragmatic hernia in dogs and cats is the traumatic one. As an anatomic alteration, surgery is the elected treatment. The purpose of this study was to evaluate videolaparoscopic surgery in the diagnostic and treatment of diaphragmatic hernia in dogs. In the first phase of the study diaphragmatic hernias were performed in dogs. One week later the dogs were summited to a laparoscopic surgery to make diagnostic and surgical correction. Laparoscopic access showed to be efficient for identification of the defect and visceral displacement and for the surgical treatment.The most common type of diaphragmatic hernia in dogs and cats is the traumatic one.As an anatomic alteration surgery is the elected treatment. The purpose of this study was to evaluate videothoracoscopic surgery in the diagnostic and treatment of diaphragmatic hernia in eight dogs. In the first phase of the study diaphragmatic hernias were performed in the dogs through laparoscopic access. One week later the dogs were summited to a thoracoscopic surgery to make diagnostic and surgical correction. Thoracoscopic access showed to be efficient for identification of the defects and visceral displacements and for the surgical treatment.

Amniotic Fluid Derived Stem Cells with a Renal Progenitor Phenotype Inhibit Interstitial Fibrosis in Renal Ischemia and Reperfusion Injury in Rats

Objectives Mesenchymal stem cells derived from human amniotic fluid (hAFSCs) are a promising source for cellular therapy, especially for renal disorders, as a subpopulation is derived from the fetal urinary tract. The purpose of this study was to evaluate if hAFSCs with a renal progenitor phenotype demonstrate a nephroprotective effect in acute ischemia reperfusion (I/R) model and prevent late stage fibrosis. Methods A total of 45 male 12-wk-old Wistar rats were divided into three equal groups;: rats subjected to I/R injury and treated with Chang Medium, rats subjected to I/R injury and treated with hAFSCs and sham-operated animals. In the first part of this study, hAFSCs that highly expressed CD24, CD117, SIX2 and PAX2 were isolated and characterized. In the second part, renal I/R injury was induced in male rats and cellular treatment was performed 6 hours later via arterial injection. Functional and histological analyses were performed 24 hours, 48 hours and 2 months after treatment using serum creatinine, urine protein to creatinine ratio, inflammatory and regeneration markers and histomorphometric analysis of the kidney. Statistical analysis was performed by analysis of variance followed by the Tukey’s test for multiple comparisons or by nonparametric Kruskal-Wallis followed by Dunn. Statistical significance level was defined as p <0.05. Results hAFSCs treatment resulted in significantly reduced serum creatinine level at 24 hours, less tubular necrosis, less hyaline cast formation, higher proliferation index, less inflammatory cell infiltration and less myofibroblasts at 48h. The treated group had less fibrosis and proteinuria at 2 months after injury. Conclusion hAFSCs contain a renal progenitor cell subpopulation that has a nephroprotective effect when delivered intra-arterially in rats with renal I/R injury, and reduces interstitial fibrosis on long term follow-up.

SOLUÇÃO HIPERSATURADA DE SAL COMO CONSERVANTE DE PERICÁRDIO CANINO UTILIZADO NA REPARAÇÃO DO MÚSCULO RETO ABDOMINAL DE RATOS WISTAR

Com a finalidade de testar a solucao hipersaturada de sal (na proporcao de 1,5g de sal comercial para 1m de agua tridestilada) como conservante, foram realizados implantes de pericardio canino mantido neste meio, no minimo por 90 dias, em lesoes provocadas no musculo reto abdominal de 24 ratos Wistar. Previamente a implantacao da membrana, a solucao foi avaliada quanto a presenca de bacterias e fungos, demonstrando resultados negativos. Durante o procedimento cirurgico, foi removido um segmento da largura completa do musculo reto abdominal direito, de 1,5cm de comprimento. A lesao produzida foi preenchida com o implante, sendo este suturado as bordas musculares com fio de nailon monofilamentar 6-0 em padrao continuo simples. Os animais operados foram subdivididos em seis grupos (I, II, III, IV, V, VI) de igual numero, sendo posteriormente sacrificados aos tres, cinco, sete,10, 15 e 30 dias do pos-operatorio, afim de se realizar as avaliacoes macroscopica e histologica da regiao do implante. Nos exames macroscopico e histologico pode-se constatar neovascularizacao no local reparado, que gradualmente foi decrescendo. Ao exame histologico foi observado a substituicao gradativa do implante por tecido conjuntivo fibroso, sem a ocorrencia de eliminacao do implante ou contaminacao. Atraves destes achados e possivel afirmar que a solucao hipersaturada de sal estudada, e um meio adequado para a conservacao de pericardio canino.

Transplante autólogo de células mononucleares da medula óssea em úlcera de córnea experimental em cães

Bone marrow (BM) mononuclear cells (MC) are a great subject in tissual regeneration. The main stem cell source to the eye is the limbus. Theses cells replace injured corneal cells, however, if the limbal stem cells are not functional, a limbal deficiency with concomitant conjunctivalization takes place. This pathological condition can be caused for several reasons, in which alkali burns are the most common. To conduct a research about transplanted BM MC presence, the cells homing and to histopathologically compare the treated and sham group, an experimental corneal ulcer model associated with MC autotransplant was used. Sixteen, male or female, stray dogs suffered experimental corneal ulceration with sodium hydroxide soaked filter discs. After the lesions, animals were submitted to subconjunctival autotransplant of previously marked BM MC. The evaluation was made by immunofluorescence on the sixth day after lesions creation and histopathology was conducted 15 days after the same procedure, when it was possible to observe that the MC grafted in the injured area, the cells did not execute the homing process and, despite the inflammatory decrease, they did not help the corneal epithelial healing process in a short term evaluation. Thus, future studies about MC transplantations in corneal ulcers are indicated.

Homoimplante ortotópico conservado, associado à terapia "soft laser" na reparação tenopatelar em cão

The aim of the present research was to develop an alternative method of treatment for tenopatellar lesions. The tenopatellar implant, preserved in glycerin 98%, was applied in 24 dogs, associated (group I - twelve dogs) or not (group II - twelve dogs) to laser therapy with gallium arsenide during the first ten post-operative days. The animals in each group were divided again in subgroups according to body weight (between 10 and 20kg or between 21 and 30kg) and according to post-operative period (60, 120 and 180 days) of evaluation. The stifle joint was immobilized through percutaneous external skeletal fixation type II for 30 post-operative days. Temporary articular immobilization provided adequate joint stability. At radiographic exams, different stages of bone resorption from the implant were verified, being more evident at 180 days post-operativelly. Microscopically, fragments of osteal trabecules of the graft, in reabsorption phase, surrounded by osteoclasts and filled with fibrovascular tissue were verified in all the groups. Newbone formation at the graft from endochondral ossification, more evident at the 180th day group, was noted. Despite the need for more studies, with a longer period of post-operatory evaluation, the implant of an orthotopic homologous tenopatellar segment preserved in glycerin 98% is suitable for replacement of tenopatellar segments in dogs weighting up to 30 kg.

Utilização de células-tronco autólogas de medula óssea na regeneração do nervo tibial de coelhos mediante técnica de tubulização com prótese de silicone

This study presents an experimental model of an acute deffect in a peripheral nerve to evaluate neural regeneration using a tubulization technique associated with the inoculation of autologous stem cells from bone marrow. A total of 12 New Zealand white rabbits underwent a bilateral dissection of the tibial nerve followed by repair with silicone tubulization. On the left tibial nerve of all animals, the tube was filled with autologous bone marrow-derived stem cells collected from the humerus. For control, using the same repair technique, the tubes were filled with a NaCl solution in the right tibial nerve. After 30 days of observation, the animals were euthanized and a histological evaluation of the collected nerve segments was performed by staining with hematoxylin-eosin, luxol fast blue, and toluidine blue. From the results it is possible to conclude that the transplanted autologous stem cells associated with the tubulization technique present an advantage in the peripheral nerve regeneration process.

Células-tronco mononucleares autólogas e proteína óssea morfogenética na cicatrização de defeitos tibiais experimentalmente induzidos em cães

Avaliou-se a utilizacao de celulas-tronco mononucleares (CTM) na cicatrizacao de defeito osseo experimental como alternativa aos metodos convencionais, analisando-se o tempo de evolucao cicatricial e a presenca dessas celulas no tecido neoformado. Foram utilizados 18 caes, separados em tres grupos (G) de seis, e de cada animal foram colhidas celulas da medula ossea (MO), contadas e analisadas para morfometria, por meio da contagem manual e mielograma. Um defeito osseo tibial foi entao criado cirurgicamente, e a lesao tratada com esponja de gelatina embebida em solucao fisiologica (G1), esponja de gelatina embebida com aspirado de MO processado (G2) e esponja de gelatina embebida com aspirado de MO processado e proteina ossea morfogenetica (rhBMP-2) (G3). A cicatrizacao foi entao avaliada por estudos radiograficos, e a presenca de CTM foi identificada por meio de marcadores nanocristais Qtracker, em microscopia com luz fluorescente, uma semana apos a intervencao cirurgica. Entre as celulas identificadas pelo marcador, foram encontradas celulas da linhagem ossea. As avaliacoes radiograficas demonstram crescimento osseo acelerado nos animais de G2 e G3. Houve diferencas significativas entre o G1 e G3 em todos os tempos estudados, e entre G1 e G2 nos tempos de 30 e 45 dias. A utilizacao de CTM adultas suplementadas ou nao com rhBMP-2 e alternativa favoravel ao crescimento osseo em defeitos experimentais agudos de tibia de caes.

Biometria ultra-sonográfica bidimensional em tempo real do globo ocular de cães

Sixty ocular globes of 30 health dogs were utilized to perform a real-time ultrasonic biometry, with the objective of getting distance measurement of intra-ocular structures. The measurements were taken from cross-sectional sagital image with the dogs in esternal recumbence, and manual containment, with the use of topical ocular anesthetic vehicle applied in the cornea. A 7.5 MHz transducer of a mechanical sector scanner without flotation pad was used. The measure averages were 3.9 ± 0.7mm (D1) distance between the midcornea to anterior capsule; 6.1 ± 1.2mm (D2) lens thickness; 10.5 ± 1.0mm (D3) lens diameter; 9.1 ± 0.4mm (D4) vitreous chamber depth; and 18.8 ± 0.9mm (D5) midcornea to retinal distance. Significant difference was observed among the right and left eyes only in D1.