MicroRNA-21 and microRNA-148a affects PTEN, NO and ROS in canine leishmaniasis

Canine Visceral leishmaniasis (CanL) poses a severe public health threat in several countries. Disease progression depends on the degree of immune response suppression. MicroRNAs (miRs) modulate mRNA translation into proteins and regulate various cellular functions and pathways associated with immune responses. MiR-21 and miR-148a can alter the parasite load and M1 macrophages are the principal cells in dogs’ leishmanicidal activity. A previous study found increased miR-21 and miR-148a in splenic leukocytes (SL) of dogs with CanL using microarray analysis and in silico analysis identified PTEN pathway targets. PTEN is involved in the immune regulation of macrophages. We measured PTEN and the production of reactive oxygen species (ROS) and nitric oxide (NO) before and after transfection SLs of dogs with CanL with mimic and inhibition of miR-21 and miR-148a. PTEN levels increased, NO and ROS decreased in SLs from dogs with CanL. Inhibition of miRNA-21 resulted in PTEN increase; in contrast, PTEN decreased after miR-148a inhibition. Nitrite (NO2) levels increased after transfection with miR-21 inhibitor but were decreased with miR-148a inhibitor. The increase in miR-21 promoted a reduction in ROS and NO levels, but miR-148a inhibition increased NO and reduced ROS. These findings suggest that miR-21 and miR-148a can participate in immune response in CanL, affecting PTEN, NO, and ROS levels

Proteína C ativada no tratamento de recém-nascido com sepse, choque e disfunção de múltiplos órgãos e sistemas: relato de caso e revisão de literatura Activated C protein in the treatment of a newborn with sepsis, shock and multiple organ dysfunction systems: case report and literature review

JUSTIFICATIVA E OBJETIVOS: A sepse grave representa a síndrome de resposta inflamatória sistêmica resultante de uma infecção, na presença de disfunção cardiovascular, síndrome do desconforto respiratório agudo ou duas ou mais disfunções orgânicas. Embora a mortalidade atribuída à sepse em crianças tenha sido reduzida de maneira significativa nas últimas décadas, a incidência de óbitos em recém-nascidos permanece elevada (20% a 40%), a despeito dos avanços em cuidados intensivos. O objetivo deste estudo foi descrever o caso de um recém-nascido com sepse, choque e disfunção de múltiplos órgãos e sistemas (DMOS) que se beneficiou do uso da proteína C ativada. RELATO DO CASO: Recém-nascido prematuro, do sexo masculino, nascido de cesariana em decorrência de ruptura prematura de membranas e sofrimento fetal agudo. Internado na UTI-Neonatal por insuficiência respiratória aguda secundária à pneumonia intra-útero. Recebeu assistência ventilatória, surfactante pulmonar exógeno e antibioticoterapia precocemente, evoluindo, no entanto, com hipertensão pulmonar persistente e choque. Houve difícil controle do quadro infeccioso, a despeito de ajustes no esquema de antibioticoterapia, evoluindo com DMOS. No 28º dia, foi iniciado o uso da proteína C ativada. O paciente evoluiu favoravelmente à medicação, com resolução das disfunções orgânicas e ausência de sangramentos. CONCLUSÕES: A proteína C ativada não pode ser prescrita de maneira rotineira no tratamento de recém-nascidos com sepse grave. No caso relatado, no entanto, acredita-se que ela tenha contribuído para a resolução das disfunções orgânicas apresentadas pelo paciente.<br>BACKGROUND AND OBJECTIVES: Severe sepsis represents the systemic inflammatory response resulting from an infection, associated with one of the following: cardiovascular organ dysfunction, acute respiratory distress syndrome or two or more organ dysfunctions. Although the mortality rate from sepsis in children has steadily decreased in the last decades, the mortality rate in newborns remains high (20% to 40%) despite the development in intensive care. The authors describe a newborn who suffered from sepsis, shock and multiple organ dysfunction syndromes (MODS) that recovered after the administration of activated C protein. CASE REPORT: A premature newborn underwent cesarean section because of a premature rupture of membranes and acute fetal distress. The newborn developed acute respiratory distress due to intrauterine pneumonia and was taken to the Neonatal Intensive Care Unit. The patient was given mechanical ventilation, exogenous pulmonary surfactant and antibiotics early in the treatment. Nevertheless, he developed persistent pulmonary hypertension and shock. The control of the infection was difficult, despite the adjustment of the antibiotics, resulting in the development of MODS. On the 28th day, activated C protein was given to the patient. The administration of the drug was successful and the patient recovered from the organ dysfunction without bleeding. CONCLUSIONS: The activated C protein can't be recommended as a routine in the treatment of newborns with severe sepsis. However, in this case, it contributed to the recovery of the organ dysfunctions presented by the patient

Vacina de DNA: uma alternativa para o tratamento do melanoma canino: revisão de literatura

O melanoma é considerado um dos tumores cutâneos de maior importância em cães devido ao seu caráter agressivo, à capacidade de produzir metástases em estágios precoces e à baixa resposta aos tratamentos não cirúrgicos. Recentemente, as vacinas de DNA apresentam-se promissoras na terapêutica do melanoma canino. O objetivo deste trabalho é apresentar uma revisão bibliográfica sobre o assunto. As vacinas de DNA são baseadas em plasmídeos que contêm o gene codificante para o antígeno alvo, expressando-o na célula do hospedeiro e apresentando, assim, vantagens em relação às vacinas tradicionais, como: facilidade de produção, estabilidade térmica, baixo custo e estimulação da resposta imune celular (linfócitos T CD8+). Devido ao sucesso limitado das terapias padrões, o estabelecimento de uma vacina de DNA efetiva aumenta a possibilidade de uma terapia promissora para o melanoma canino, podendo trazer novas expectativas aos animais portadores dessa neoplasia

Repair of lamellar scleral lesions in dogs with preserved equine renal capsule: short report Reparação de lesões lamelares esclerais em cães com cápsula renal de eqüino preservada: relato curto

PURPOSE: To evaluate the u se of equine renal capsule preserved in glycerin to repair lamellar scleral lesions in dogs. METHODS: Twelve healthy mongrel dogs, male and female, weighing 12 kg were used. The study was both clinical and morphological, and performed on the first, third, seventh, 15th, 30th and 60th day after surgery. Temporal canthotomy was performed after standard preoperative and general anesthesia. Conjunctival and scleral square incisions of 0.5x0.5 cm were carried out in a one o'clock position, near the limbus. A fragment of hydrated biological prosthesis, of the same shape, was sutured with 7-0 vicryl® in an interrupted suture. RESULTS: The clinical evaluation showed blepharospasm/photophobia until the 7th day after surgery. Conjunctival edema appeared up to the 5th day after surgery. Mucoid ocular discharge was sustained until the 10th day after surgery. Hyperemia was observed until the end of the evaluation period. There were no signs of graft extrusion in all animals. The anterior and posterior segments did not show clinical signs of inflammation. The optical microscopy morphological evaluation showed an inflammatory exudation with acute aspects in the early and intermediate periods, and inflammatory exudation with chronic aspects in the late periods. There was incorporation of the implant by the recipient's sclera. CONCLUSION: These results suggest that the equine renal preserved capsule could be a useful alternative tissue to repair lamellar corneal lesions in dogs and humans.<br>OBJETIVO: Avaliar o uso da cápsula renal de eqüino preservada em glicerina 98% no reparo de lesões lamelares esclerais em cães. MÉTODOS: Foram utilizados 12 cães, machos e fêmeas, com peso médio de 12kg. Foram realizadas avaliações clínica e morfológica aos 1, 3, 7, 15, 30 e 60 dias de pós-operatório. Após anestesia geral e procedimentos padrões de preparo do campo operatório, foi realizada cantotomia temporal, seguida de incisão conjutival e escleral com área de 0,5x0,5 cm na posição de 1hora, próxima ao limbo. Em seguida, um fragmento de mesma dimensão de cápsula renal de eqüino preservada em glicerina, previamente hidratado em solução salina, foi aplicado ao defeito escleral criado sendo fixado com pontos simples isolados com vicryl 7-0®. RESULTADOS: A avaliação clínica revelou blefaroespasmo/fotofobia até o sétimo dia de pós-operatório. Foi observado edema conjuntival até o quinto dia, acompanhado de secreção ocular mucóide, que persistiu até o décimo dia de pós-operatório. Não foram observados sinais clínicos de rejeição do enxerto em todos os animais, em todos os períodos avaliados. Os segmentos anterior e posterior do bulbo ocular não apresentaram sinais de inflamação. A análise morfológica revelou exsudação inflamatória aguda nos períodos precoces e intermediários da avaliação e inflamação crônica nos períodos tardios da observação. Houve incorporação do enxerto ao leito receptor. CONCLUSÃO: Os resultados sugerem que a cápsula renal de eqüino preservada pode ser mais uma alternativa de membrana biológica para o reparo de lesões esclerais lamelares em cães e no homem

Apoptosis in T lymphocytes from spleen tissue and peripheral blood of L. (L.) chagasi naturally infected dogs

Dogs are the main domestic reservoirs of L. (L) chagasi. Once in the vertebrate host, the parasite may cause visceral leishmaniasis, which can also be transmitted to humans. Infected symptomatic dogs show disorganization in the white pulp in spleen tissue and a reduction in T lymphocytes in peripheral blood. To investigate whether apoptosis is involved in white pulp disorganization and diminished T cell counts in peripheral blood, apoptotic T cells from the spleen and peripheral blood of dogs naturally infected with L. (L) chagasi and presenting clinical manifestations were quantified and compared with healthy dogs. Thirteen symptomatic adult dogs infected by L. (L) chagasi and six healthy dogs from a nonendemic area (controls) were included in the study. Samples from spleen and peripheral blood were used to quantify apoptosis in CD3 lymphocytes by flow cytometry using Anexin V and Multicas-pase kits; the results were compared using the Mann Whitney test. The percentage of total T cells was lower in Leishmania infected dogs compared to healthy controls (P<0.05). Apoptosis levels in T cells from PBMC and spleen were higher in infected dogs than in controls (P < 0.05). The least squares method test was used to determine the effect between the degree of structural organization of spleen white pulp and the percentage of apoptosis in the spleen. A significant effect on the level of white pulp morphological disorganization and percentage of apoptosis in spleen T cells was observed (F = 20.45: P = 0.0014). These data suggest that apoptosis is an important for the immunopathogenesis of canine visceral leishmaniasis. (C) 2011 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Concentration of TNF-α and IL-1β during CanL.

TNF-α (A) and IL-1β (B) were quantified, by capture ELISA, in SL supernatants cultures from control (dogs without L. infantum infection) and CanL (dogs naturally infected by L. infantum) groups. Splenic leukocytes were maintained in culture for 48 h at 37°C and 5% CO2, without treatment. Bars represent concentration median values of TNF-α (2.51 and 10.81) and IL-1β (0.0 and 15.70), 25th percentile interquartile range of TNF-α (1.18 and 6.49) and IL-1β (0.0 and 6.238) and 75th percentile interquartile range of TNF-α (12.89 and 24.80) and IL-1β (0.0 and 15.70) from control and CanL groups, respectively. Symbols represent data from individual animals. Asterisks represent significant differences (Mann-Whitney test, p = 0.04 (A and B).</p

Table6_MicroRNA-21 and microRNA-148a affects PTEN, NO and ROS in canine leishmaniasis.DOCX

Canine Visceral leishmaniasis (CanL) poses a severe public health threat in several countries. Disease progression depends on the degree of immune response suppression. MicroRNAs (miRs) modulate mRNA translation into proteins and regulate various cellular functions and pathways associated with immune responses. MiR-21 and miR-148a can alter the parasite load and M1 macrophages are the principal cells in dogs’ leishmanicidal activity. A previous study found increased miR-21 and miR-148a in splenic leukocytes (SL) of dogs with CanL using microarray analysis and in silico analysis identified PTEN pathway targets. PTEN is involved in the immune regulation of macrophages. We measured PTEN and the production of reactive oxygen species (ROS) and nitric oxide (NO) before and after transfection SLs of dogs with CanL with mimic and inhibition of miR-21 and miR-148a. PTEN levels increased, NO and ROS decreased in SLs from dogs with CanL. Inhibition of miRNA-21 resulted in PTEN increase; in contrast, PTEN decreased after miR-148a inhibition. Nitrite (NO2) levels increased after transfection with miR-21 inhibitor but were decreased with miR-148a inhibitor. The increase in miR-21 promoted a reduction in ROS and NO levels, but miR-148a inhibition increased NO and reduced ROS. These findings suggest that miR-21 and miR-148a can participate in immune response in CanL, affecting PTEN, NO, and ROS levels.</p