Replication of Rocio virus in primary cultures of mouse neural cells

ABSTRACT Objective This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. Methods Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. Results Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. Conclusion The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.

Digital radiography as an alternative method in the evaluation of bone density in uremic rats / Radiografia digital como método alternativo na avaliação da densidade óssea em ratos urêmicos

ABSTRACT Introduction: Digital radiography (DRx) may provide a suitable alternative to investigate mineral and bone disorder (MBD) and loss of bone density (BD) in rodent models of chronic kidney disease (CKD). The objective of this study was to use DRx to evaluate BD in CKD rats, and to evaluate the correlation between DRx findings and serum MBD markers and bone histomorphometry. Methods: Uremia was induced by feeding Wistar rats an adenine-enriched diet (0.75% for 4 weeks/0.10% for 3 weeks); outcomes were compared to a control group at experimental weeks 3, 4, and 7. The following biochemical markers were measured: creatinine clearance (CrC), phosphate (P), calcium (Ca), fractional excretion of P (FeP), alkaline phosphatase (ALP), fibroblast growth factor-23 (FGF-23), and parathyroid hormone (PTH). DRx imaging was performed and histomorphometry analysis was conducted using the left femur. Results: As expected, at week 7, uremic rats presented with reduced CrC and higher levels of P, FeP, and ALP compared to controls. DRx confirmed the lower BD in uremic animals (0.57±0.07 vs. 0.68 ± 0.06 a.u.; p = 0.016) compared to controls at the end of week 7, when MBD was more prominent. A severe form of high-turnover bone disease accompanied these biochemical changes. BD measured on DRx correlated to P (r=-0.81; p = 0.002), ALP (r = -0.69, p = 0.01), PTH (r = -0.83, p = 0.01), OS/BS (r = -0.70; p = 0.02), and ObS/BS (r = -0.70; p = 0.02). Conclusion: BD quantified by DRx was associated with the typical complications of MBD in CKD and showed to be viable in the evaluation of bone alterations in CKD.

RESUMO Introdução: A radiografia digital (RxD) pode representar uma alternativa adequada para investigar o distúrbio mineral e ósseo (DMO) e a perda de densidade óssea (DO) em modelos de roedores da doença renal crônica (DRC). O objetivo deste estudo foi utilizar a RxD para avaliar a DO em ratos com DRC, e avaliar a correlação entre os achados da RxD e marcadores séricos de DMO e histomorfometria óssea. Métodos: A uremia foi induzida pela alimentação de ratos Wistar com dieta enriquecida com adenina (0,75% por 4 semanas/0,10% por 3 semanas); os resultados foram comparados com um grupo controle nas semanas experimentais 3, 4 e 7. Os seguintes marcadores bioquímicos foram medidos: clearance de creatinina (CCr), fosfato (P), cálcio (Ca), fração excretada de P (FeP), fosfatase alcalina (ALP), fator de crescimento de fibroblastos-23 (FGF-23) e paratormônio (PTH). A imagem da RxD foi obtida e a análise histomorfométrica foi realizada com o fêmur esquerdo. Resultados: como esperado, na semana 7, os ratos urêmicos apresentaram redução do CCr e níveis mais altos de P, FeP e ALP em comparação aos controles. A RxD confirmou a menor DO em animais urêmicos (0,57 ± 0,07 vs. 0,68 ± 0,06 u.a.; p = 0,016) em comparação aos controles no final da semana 7, quando a DMO foi mais proeminente. Uma forma grave de doença óssea de alta renovação celular acompanhou essas mudanças bioquímicas. A DO, medida na RxD foi correlacionada a P (r = -0,81; p = 0,002), ALP (r = -0,69, p = 0,01), PTH (r = -0,83, p = 0,01), OS/BS (r = -0,70 p = 0,02) e Ob.S/BS (r = -0,70; p = 0,02). Conclusão: A DO quantificada por RxD esteve associada às complicações típicas da DMO na DRC e mostrou-se viável na avaliação de alterações ósseas na DRC.

Delayed alveolar bone repair and osteonecrosis associated with Zoledronic Acid therapy in rats: macroscopic, microscopic and molecular analysis

Abstract Objective This study aims to evaluate bone repair and the development of the medication related osteonecrosis of the jaw (MRONJ) associated with the use of zoledronic acid in Wistar rats. Methodology 48 male Wistar rats were divided into four groups: ZA, treated with intraperitoneal zoledronic acid, 0.6 mg/kg every 28 days, totaling five doses; control (C), treated with 0.9% sodium chloride; ZA-surgical (SZA) and C-surgical (SC), submitted to extraction of the right upper molars 45 days after the first application. Alveolar bone repair was evaluated by macroscopic and histological analysis. Protein expression evaluations were performed by qPCR. Results Macroscopic evaluation showed that 91.66% (11) of the animals in the SZA group and 41.66% (5) from the SC group presented solution of epithelium continuity (P<0.05). All animals in the SZA group and none in the SC group had bone sequestration. The area of osteonecrosis was higher in the SZA group than in the SC group (P<0.05). In molecular evaluation, the SZA group presented changes in the expression of markers for osteoclasts, with increased RANK and RANKL, and a decrease in OPG. Conclusion The results highlighted strong and evident interference of zoledronic acid in bone repair of the socket, causing osteonecrosis and delayed bone remodeling.

Inflammatory lung injury in rabbits: effects of high-frequency oscillatory ventilation in the prone position / Lesão inflamatória pulmonar em coelhos: efeitos da ventilação oscilatória de alta frequência em posição prona

ABSTRACT Objective: To compare the effects that prone and supine positioning during high-frequency oscillatory ventilation (HFOV) have on oxygenation and lung inflammation, histological injury, and oxidative stress in a rabbit model of acute lung injury (ALI). Methods: Thirty male Norfolk white rabbits were induced to ALI by tracheal saline lavage (30 mL/kg, 38°C). The injury was induced during conventional mechanical ventilation, and ALI was considered confirmed when a PaO2/FiO2 ratio < 100 mmHg was reached. Rabbits were randomly divided into two groups: HFOV in the supine position (SP group, n = 15); and HFOV with prone positioning (PP group, n = 15). For HFOV, the mean airway pressure was initially set at 16 cmH2O. At 30, 60, and 90 min after the start of the HFOV protocol, the mean airway pressure was reduced to 14, 12, and 10 cmH2O, respectively. At 120 min, the animals were returned to or remained in the supine position for an extra 30 min. We evaluated oxygenation indices and histological lung injury scores, as well as TNF-α levels in BAL fluid and lung tissue. Results: After ALI induction, all of the animals showed significant hypoxemia, decreased respiratory system compliance, decreased oxygenation, and increased mean airway pressure in comparison with the baseline values. There were no statistically significant differences between the two groups, at any of the time points evaluated, in terms of the PaO2 or oxygenation index. However, TNF-α levels in BAL fluid were significantly lower in the PP group than in the SP group, as were histological lung injury scores. Conclusions: Prone positioning appears to attenuate inflammatory and histological lung injury during HFOV in rabbits with ALI.

RESUMO Objetivo: Comparar os efeitos das posições prona e supina durante ventilação oscilatória de alta frequência (VOAF) sobre oxigenação e inflamação pulmonar, lesão histológica e estresse oxidativo em um modelo de lesão pulmonar aguda (LPA) em coelhos. Métodos: Trinta coelhos Norfolk machos brancos foram submetidos à LPA por meio de lavagem traqueal com salina (30 ml/kg, 38°C). A lesão foi induzida durante a ventilação mecânica convencional, e a LPA foi considerada confirmada na presença de relação PaO2/FiO2 < 100 mmHg. Os coelhos foram aleatoriamente divididos em dois grupos: VOAF em posição supina (grupo PS, n = 15); e VOAF em posição prona (grupo PP, n = 15). Para a VOAF, a pressão média das vias aéreas foi inicialmente estabelecida em 16 cmH2O. No 30º, 60º e 90º min após o início do protocolo de VOAF, a pressão média das vias aéreas foi reduzida para 14, 12 e 10 cmH2O, respectivamente. No 120º min, os animais foram recolocados ou permaneceram na posição supina por mais 30 min. Foram avaliados os índices de oxigenação e escores histológicos de lesão pulmonar, bem como os níveis de TNF-α em lavado broncoalveolar e tecido pulmonar. Resultados: Após a indução da LPA, todos os animais apresentaram hipoxemia significativa, diminuição da complacência do sistema respiratório, diminuição da oxigenação e aumento da pressão média das vias aéreas em comparação aos valores basais. Não houve diferenças estatisticamente significativas entre os dois grupos, em nenhum dos momentos avaliados, quanto a PaO2 e índice de oxigenação. Entretanto, os níveis de TNF-α no lavado broncoalveolar foram significativamente menores no grupo PP que no grupo PS, assim como os escores histológicos de lesão pulmonar. Conclusões: A posição prona parece atenuar a lesão pulmonar inflamatória e histológica durante a VOAF em coelhos com LPA.

Anti-inflammatory effect of lycopene on endotoxin-induced uveitis in rats / Efeito anti-inflamatório do licopeno na uveíte induzida por endotoxina em ratos

ABSTRACT Purpose: We evaluated the efficacy of lycopene, a dietary carotenoid and potent antioxidant, against ocular inflammation and oxidative stress in an experimental uveitis model. Methods: Endotoxin-induced uveitis (EIU) was induced in Sprague-Dawley rats by a single subcutaneous injection of 200 μg lipopolysaccharide (LPS). Induction of EIU was preceded by daily intraperitoneal injection of 10 mg/kg lycopene for three consecutive days (Lycopene + LPS group) or equivolume vehicle (Vehicle + LPS group). A positive control group received 1 mg/kg dexamethasone pretreatment (DEX + LPS), and a negative control group received daily vehicle injection but no LPS (Vehicle Control). Twenty-four hours after LPS or final vehicle administration, eyes were enucleated, and aqueous humor was collected for measurement of the number of infiltrating cells, total protein concentration, and levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and oxidative stress markers. Inflammatory response severity was compared among groups clinically and histopathologically. Results: Infiltrating cell number, total protein concentration, and NO, TNF-α, and IL-6 levels were significantly elevated in the aqueous humor of Vehicle + LPS group rats compared to Vehicle Controls. Compared to the Vehicle + LPS group, lycopene pretreatment significantly reduced aqueous humor concentrations of oxidative stress markers, NO (0.29 ± 0.1 μM vs. 0.19 ± 0.1 μM, p=0.003), TNF-α (71.0 ± 22.3 ng/ml vs. 50.1 ± 2.1 ng/ml, p=0.043), and IL-6 (121.6 ± 3.0 pg/ml vs. 111.1 ± 5.6 pg/ml, p=0.008). Inflammatory score was also reduced (2.0 ± 0.0 vs. 0.4 ± 0.5, p=0.001). Lycopene reduced the infiltrating cell count and protein concentration, but differences did not reach significance. Most lycopene effects were equivalent to dexamethasone. Conclusions: Lycopene may aid in the clinical management of uveitis by suppressing inflammation and oxidative stress.

RESUMO Objetivo: Avaliamos o efeito do licopeno, um carotenóide dietético e um potente anti-oxidante, sobre a inflamação ocular e estresse oxidativo em modelo de uveíte experimental. Métodos: Uveíte foi induzida por endotoxina (EIU) em ratos Sprague-Dawley por uma única injeção subcutânea de 200 ug de lipopolissacárido (LPS). A indução de EIU foi precedida por injeção intraperitoneal de licopeno em uma dose de 10 mg/kg (grupo LPS + Licopeno) ou veículo de mesmo volume (grupo LPS + Veículo), durante 3 dias consecutivos. O grupo controle positivo recebeu uma dose de 1 mg/kg de Dexametasona (grupo DEX + LPS) e o grupo controle negativo recebeu doses diárias de veículo mas sem LPS (grupo Controle Veículo). Vinte e quatro horas após a administração do LPS, os olhos foram enucleados, humor aquoso foi recolhido, e o número de células infiltrativas, a concentração de proteína, assim como os níveis de óxido nítrico (NO), fator de necrose tumoral α (TNF-α), interleucina-6 e marcadores de estresse oxidativo foram determinados no humor aquoso. Além disso, a resposta inflamatória foi avaliada clinicamente e histologicamente. Resultados: As células infiltrativas, concentração de proteína, o NO, TNF-α, interleucina-6 foram significativamente elevados no humor aquoso de ratos do grupo Grupo LPS + Veículo quando comparados ao Grupo Controle Veículo. O tratamento com licopeno diminuiu significativamente estes aumentos. Comparado ao Grupo LPS + Veículo, o licopeno reduziu significativamente as concentrações no humor aquoso dos marcadores de estresse oxidativo e NO (de 0,29 ± 0,1 μM para 0,19 ± 0,1 μM, p=0,003), o TNF-α (de 71,0 ± 22,3 ng/ml para 50,1 ± 2,1 ng/ml, p=0,043), interleucina-6 (de 121,6 ± 3,0 pg/ml para 111,1 ± 5,6 pg/ml, p=0,008). Do mesmo modo, o aumento do número de células infiltrativas no tecido uveal em seções histológicas foi significativamente inibido pelo licopeno, a pontuação inflamatória diminuiu de 2,0 ± 0,0 para 0,4 ± 0,5, p=0,001. Embora, não tenha sido estatisticamente significativo, o licopeno reduziu a contagem de células infiltrativas e a concentração de proteínas no humor aquoso. Conclusões: Estes resultados sugerem que o licopeno pode ter efeitos benéficos no tratamento da inflamação ocular, através dos seus efeitos anti-inflamatórios e antioxidantes.

Reposição de vitamina B12 reduz comportamento depressivo induzido em ratos jovens / Vitamin B12 replacement therapy reduces induced depressive behavior in young rats

OBJETIVOS: Avaliar, em modelo animal, se a depleção suave de vitamina B12, anterior ao desenvolvimento de anemia, induz à depressão; e se a suplementação com vitamina B12 em animais jovens pode atuar como medida preventiva da depressão. MÉTODOS: Foram utilizados ratos Wistar divididos em grupo controle (n=11) e grupo B12 (n=10). O grupo B12 recebeu suplementação de vitamina B12 na água de beber, ao longo de todo o estudo. Na fase 1, os animais dos dois grupos receberam por seis semanas dieta adicionada de pectina (50g/kg da ração), para induzir à depleção de vitamina B12. Após esse período, foi aplicado o Teste de Porsolt para indução e avaliação do estado depressivo. Foi realizado também um hemograma para pesquisa de anemia. Na fase 2 (com duração de quatro semanas), a pectina foi removida da ração e os mesmos testes foram aplicados novamente no final do período. RESULTADOS: Durante as duas fases do estudo o número de hemácias, o hematócrito e a concentração de hemoglobina mantiveram-se normais, ou seja, os ratos não desenvolveram anemia. Os resultados do Teste de Nado Forçado ao final da fase 1 mostram que, em relação ao grupo controle, o grupo suplementado apresentou tempo de desistência menor (0,44±0,32 vs. 0,75±0,18 minutos, p=0,024) e tempo de natação maior (4,64±0,27 vs. 4,32±0,28 minutos, p=0,013), indicando redução do estado depressivo com a reposição de vitamina B12. Na comparação entre grupos no final da fase 2 não houve diferença significativa em nenhum dos componentes do Teste de Nado Forçado. CONCLUSÕES: A depleção suave de vitamina B12 na dieta, em nível não indutor de anemia, favoreceu o estado depressivo em ratos jovens, enquanto a sua suplementação na situação de depleção reverteu esse quadro. Em condições de nutrição adequada, entretanto, a suplementação dessa vitamina não exerceu efeito sobre o estado depressivo. Estes resultados estimulam a realização de mais estudos que aprofundem a avaliação das relações entre vitamina B12 e depressão em jovens. Além disso, este estudo também abre perspectivas para um novo modelo experimental de depressão, induzida por depleção de vitamina B12.

AIMS: To assess whether mild vitamin B12 deficiency induces depression prior to the development of anemia, and whether vitamin B12 supplementation can act as a preventive measure against depression in young rats. METHODS: Wistar rats were divided into control group (n=11) and B12 group (n=10). The B12 group received vitamin B12 supplementation in drinking water throughout the study. In Phase 1, all animals received a pectin-supplemented diet (50g/kg) for six weeks to induce vitamin B12 depletion. After that, the Porsolt test was applied for induction and evaluation of depressive state and blood was collected for a complete blood count. In Phase 2, which lasted two weeks, pectin was removed from the diet and the same tests were applied again at the end. RESULTS: In both phases, erythrocyte count, hematocrit level, and hemoglobin concentration were normal, i.e., the rats did not develop anemia. The forced swim test results at the end of Phase 1 show that the B12 group exhibited shorter immobility time than the control group (0.44±0.32 vs. 0.75±0.18 minutes, p=0.024) and longer swimming time (4.64±0.27 vs. 4.32±0.28 minutes, p=0.013), indicating reduction of depressive state with vitamin B12 replacement therapy. When the groups were compared at the end of Phase 2, there was no significant difference in any of the forced swim test components. CONCLUSIONS: Mild vitamin B12 deficiency, at a level that did not induce anemia, led to depressive state in young rats where vitamin B12 supplementation reversed the effects of vitamin depletion. Under normal nutritional circumstances, however, vitamin B12 supplementation did not have any effect on depressive state. These findings encourage further studies to investigate the associations between vitamin B12 and depression in young individuals. Moreover, this study also presents perspectives for a new experimental model of depression induced by vitamin B12 depletion.

Caso de hemianopsia reversível no pós-parto decorrente de síndrome da encefalopatia reversível posterior (PRES) em gestante com eclâmpsia tardia / Reversible hemianopsia in postpartum due to posterior reversible encephalopathy syndrome in pregnant with late eclampsia

Resumo Objetivos: Descrever um caso de Síndrome da Encefalopatia Reversível Posterior em gestante diagnosticada com eclâmpsia tardia, bem como seu manejo clínico. Descrição do caso: Paciente feminina, 34 anos, em sua terceira gestação, iniciou com aumento dos níveis tensionais durante o trabalho de parto e, após onze dias de puerpério, apresentou quadro de diminuição da acuidade visual à direita, seguida de crise convulsiva e subsequente perda parcial da visão do olho direito. Após a realização de exames de imagem e descartada a possibilidade de acidente vascular encefálico, a paciente foi diagnosticada com Síndrome da Encefalopatia Reversível Posterior (PRES). Instituído o manejo clínico das crises convulsivas e hipertensivas, houve remissão completa dos sintomas e reversão do quadro clínico inicial. Conclusões: Uma vez adequadamente diagnosticada e tratada, a Síndrome da Encefalopatia Reversível Posterior pode apresentar evolução satisfatória, especialmente quando associada a um fator desencadeado agudamente, como a eclâmpsia.

Abstract Objectives: To describe a case of Posterior Reversible Encephalopathy Syndrome diagnosed in pregnant women with late-eclampsia, as well as its clinical management. Case description: A 34 years old patient in her third pregnancy had started with high blood pressure levels during labor; after eleven days postpartum, she presented a decreased right visual acuity; subsequently one episode of seizure followed by partial loss of vision in the right eye. After conducting tests and ruled out stroke, the patient was diagnosed as Posterior Reversible Encephalopathy Syndrome (PRES). Established the clinical management of seizures and hypertensive crisis, there was complete remission of symptoms and reversal of the initial clinical picture. Conclusion: Once properly diagnosed and treated, the Posterior Reversible Encephalopathy Syndrome can present satisfactory progress, especially when associated with an acutely triggered factor, as eclampsia.

Chronic intermittent hypoxia increases encoding pigment epithelium-derived factor gene expression, although not that of the protein itself, in the temporal cortex of rats.

OBJECTIVE: Obstructive sleep apnea syndrome is mainly characterized by intermittent hypoxia (IH) during sleep, being associated with several complications. Exposure to IH is the most widely used animal model of sleep apnea, short-term IH exposure resulting in cognitive and neuronal impairment. Pigment epithelium-derived factor (PEDF) is a hypoxia-sensitive factor acting as a neurotrophic, neuroprotective, and antiangiogenic agent. Our study analyzed performance on learning and cognitive tasks, as well as PEDF gene expression and PEDF protein expression in specific brain structures, in rats exposed to long-term IH. METHODS: Male Wistar rats were exposed to IH (oxygen concentrations of 21-5%) for 6 weeks-the chronic IH (CIH) group-or normoxia for 6 weeks-the control group. After CIH exposure, a group of rats were allowed to recover under normoxic conditions for 2 weeks (the CIH+N group). All rats underwent the Morris water maze test for learning and memory, PEDF gene expression and PEDF protein expression in the hippocampus, frontal cortex, and temporal cortex being subsequently assessed. RESULTS: The CIH and CIH+N groups showed increased PEDF gene expression in the temporal cortex, PEDF protein expression remaining unaltered. PEDF gene expression and PEDF protein expression remained unaltered in the frontal cortex and hippocampus. Long-term exposure to IH did not affect cognitive function. CONCLUSIONS: Long-term exposure to IH selectively increases PEDF gene expression at the transcriptional level, although only in the temporal cortex. This increase is probably a protective mechanism against IH-induced injury.

OBJETIVO: A síndrome da apneia obstrutiva do sono caracteriza-se principalmente por episódios de hipóxia intermitente (HI) durante o sono e associa-se a diversas complicações. A exposição à HI é o mais usado modelo animal de apneia do sono, e protocolos de curta duração causam diversos prejuízos cognitivos e neuronais. Pigment epithelium-derived factor (PEDF, fator derivado do epitélio pigmentado) é um fator neurotrófico, neuroprotetor e antiangiogênico sensível à hipóxia celular. Nosso estudo analisou o desempenho em tarefas cognitivas e de aprendizagem, bem como a expressão do gene PEDF e da proteína PEDF em estruturas cerebrais específicas em ratos expostos a HI de longa duração. MÉTODOS: Ratos Wistar foram expostos a HI (21-5% de oxigênio) durante 6 semanas - o grupo HI crônica (HIC) - ou a normóxia durante 6 semanas - o grupo controle. Após a exposição à HIC, um grupo de ratos foi exposto a normóxia durante 2 semanas (o grupo HIC+N). Todos os animais foram submetidos ao labirinto aquático de Morris para avaliação de memória e aprendizado; avaliou-se também a expressão do gene PEDF e da proteína PEDF no hipocampo e nos córtices frontal e temporal. RESULTADOS: Os grupos HIC e HIC+N apresentaram um aumento de expressão do gene PEDF no córtex temporal, porém sem aumento dos níveis proteicos. A expressão do gene PEDF e da proteína PEDF manteve-se inalterada nas demais estruturas. A exposição de longa duração à HI não afetou a função cognitiva. CONCLUSÕES: A exposição de longa duração à HI aumenta seletivamente a expressão do gene PEDF ao nível transcricional, embora apenas no córtex temporal. Esse aumento é provavelmente um mecanismo de proteção contra a HI.

Which is the best route of administration for cell therapy in experimental model of small-for size syndrome in rats?

PURPOSE: To evaluate which is the best route of administration for cell therapy in experimental rat model of small-for size syndrome. METHODS: A total of 40 rats underwent partial hepatectomy (70%) that induces the small-for-size syndrome and were divided into four groups of route administration: intravenous, intraperitoneal, enteral and tracheal. The small-for-size syndrome model was designed with extended partial hepatectomy (70%). The animals were divided into four groups of routes administration: intravenous (n=10) - intravenously through the dorsal vein of the penis; intraperitoneal (n=10) - intraperitoneally in the abdominal cavity; enteral (n=10) - oroenteral with the placement of a number 4 urethral probe and maintained at third duodenal portion; tracheal (n=10) - after tracheal intubation. We track the animals and monitor them for 21 days; during this follow-up we evaluated the result of cell therapy application tracking animals using ultrasound, radiography and PET-scan. Statistical analysis was performed using GraphPad Prism Software(r). Differences were considered significant with the p<0.05. Data are presented as the median and variation for continuous variables. Comparisons between groups were made using analysis of the imaging test by the researchers. RESULTS: All four groups underwent partial hepatectomy of 70% liver tissue targeting the same weight of resected liver. Initially the PET-scan tests showed similarity in administered cells by different routes. However, in few days the route of intravenous administration showed to be the most appropriated to lead cells to the liver followed by enteral. The tracheal and peritoneal routes were not as much successful for this goal. CONCLUSION: The intravenous route is the best one to cell therapy in experimental rat model of small-for size-syndrome. .

Chronic intermittent hypoxia increases encoding pigment epithelium-derived factor gene expression, although not that of the protein itself, in the temporal cortex of rats, / Hipóxia intermitente crônica aumenta a expressão gênica, mas não proteica, de pigment epithelium-derived factor no córtex temporal de ratos

Objective: Obstructive sleep apnea syndrome is mainly characterized by intermittent hypoxia (IH) during sleep, being associated with several complications. Exposure to IH is the most widely used animal model of sleep apnea, short-term IH exposure resulting in cognitive and neuronal impairment. Pigment epithelium-derived factor (PEDF) is a hypoxia-sensitive factor acting as a neurotrophic, neuroprotective, and antiangiogenic agent. Our study analyzed performance on learning and cognitive tasks, as well as PEDF gene expression and PEDF protein expression in specific brain structures, in rats exposed to long-term IH. Methods: Male Wistar rats were exposed to IH (oxygen concentrations of 21-5%) for 6 weeks-the chronic IH (CIH) group-or normoxia for 6 weeks-the control group. After CIH exposure, a group of rats were allowed to recover under normoxic conditions for 2 weeks (the CIH+N group). All rats underwent the Morris water maze test for learning and memory, PEDF gene expression and PEDF protein expression in the hippocampus, frontal cortex, and temporal cortex being subsequently assessed. Results: The CIH and CIH+N groups showed increased PEDF gene expression in the temporal cortex, PEDF protein expression remaining unaltered. PEDF gene expression and PEDF protein expression remained unaltered in the frontal cortex and hippocampus. Long-term exposure to IH did not affect cognitive function. Conclusions: Long-term exposure to IH selectively increases PEDF gene expression at the transcriptional level, although only in the temporal cortex. This increase is probably a protective mechanism against IH-induced injury. .

Objetivo: A síndrome da apneia obstrutiva do sono caracteriza-se principalmente por episódios de hipóxia intermitente (HI) durante o sono e associa-se a diversas complicações. A exposição à HI é o mais usado modelo animal de apneia do sono, e protocolos de curta duração causam diversos prejuízos cognitivos e neuronais. Pigment epithelium-derived factor (PEDF, fator derivado do epitélio pigmentado) é um fator neurotrófico, neuroprotetor e antiangiogênico sensível à hipóxia celular. Nosso estudo analisou o desempenho em tarefas cognitivas e de aprendizagem, bem como a expressão do gene PEDF e da proteína PEDF em estruturas cerebrais específicas em ratos expostos a HI de longa duração. Métodos: Ratos Wistar foram expostos a HI (21-5% de oxigênio) durante 6 semanas - o grupo HI crônica (HIC) - ou a normóxia durante 6 semanas - o grupo controle. Após a exposição à HIC, um grupo de ratos foi exposto a normóxia durante 2 semanas (o grupo HIC+N). Todos os animais foram submetidos ao labirinto aquático de Morris para avaliação de memória e aprendizado; avaliou-se também a expressão do gene PEDF e da proteína PEDF no hipocampo e nos córtices frontal e temporal. Resultados: Os grupos HIC e HIC+N apresentaram um aumento de expressão do gene PEDF no córtex temporal, porém sem aumento dos níveis proteicos. A expressão do gene PEDF e da proteína PEDF manteve-se inalterada nas demais estruturas. A exposição de longa duração à HI não afetou a função cognitiva. Conclusões: A exposição de longa duração à HI aumenta seletivamente a expressão do gene PEDF ao nível transcricional, embora apenas no córtex temporal. Esse aumento é provavelmente um mecanismo de proteção contra a HI. .

Development of a subcutaneous endometriosis rat model

PURPOSE: To present a rat model of subcutaneous endometriosis for the study of pathophysiology and the effects of drugs. METHODS: Fifty three-month-old female Wistar rats (Rattus norvergicus) were distributed into one control group and four treatment groups: estradiol (2.5; 5; 10mg/kg sc), medroxyprogesterone acetate (0.5; 2; 5mg/kg sc), triptorelin pamoate (0.18; 0.56mg/kg sc) and acetylsalicylic acid (3mg/kg per os). The animals were autoimplanted subcutaneously with 4x4-mm uterine fragments to induce endometriosis. The endometriomas were measured on days 1, 7, 14 and 21. The relative dry and wet weights of the endometrioma were used to evaluate response to the drug. Endometrial -like tissue was confirmed by histology. The greatest weight gain was observed on day 14 (relative wet weight: 29.1 ± 6.7mg%, relative dry weight: 5.3 ± 0.9mg %). Treatments were administered between day 5 and day 14. RESULTS: The relative wet weight of the hemiuterus in the 10mg/kg estradiol group differed significantly from control and the other two estradiol groups (p=0.0001). In the medroxyprogesterone acetate group the weight decreased significantly but this decrease was not dose-dependent. Weight reduction was also significant in the triptorelin pamoate and the acetylsalicylic acid groups. CONCLUSION: The model of subcutaneous endometriosis is reproducible, low-cost and easy to perform, and suitable for the study of pathophysiology and the effects of drugs. .

Which is the best route of administration for cell therapy in experimental model of small-for size syndrome in rats?

PURPOSE: To evaluate which is the best route of administration for cell therapy in experimental rat model of small-for size syndrome. METHODS: A total of 40 rats underwent partial hepatectomy (70%) that induces the small-for-size syndrome and were divided into four groups of route administration: intravenous, intraperitoneal, enteral and tracheal. The small-for-size syndrome model was designed with extended partial hepatectomy (70%). The animals were divided into four groups of routes administration: intravenous (n=10) - intravenously through the dorsal vein of the penis; intraperitoneal (n=10) - intraperitoneally in the abdominal cavity; enteral (n=10) - oroenteral with the placement of a number 4 urethral probe and maintained at third duodenal portion; tracheal (n=10) - after tracheal intubation. We track the animals and monitor them for 21 days; during this follow-up we evaluated the result of cell therapy application tracking animals using ultrasound, radiography and PET-scan. Statistical analysis was performed using GraphPad Prism Software®. Differences were considered significant with the p<0.05. Data are presented as the median and variation for continuous variables. Comparisons between groups were made using analysis of the imaging test by the researchers. RESULTS: All four groups underwent partial hepatectomy of 70% liver tissue targeting the same weight of resected liver. Initially the PET-scan tests showed similarity in administered cells by different routes. However, in few days the route of intravenous administration showed to be the most appropriated to lead cells to the liver followed by enteral. The tracheal and peritoneal routes were not as much successful for this goal.(AU)

Comparison of the effects of subconjunctival and topical anti-VEGF therapy (bevacizumab) on experimental corneal neovascularization / Comparação do efeito da terapia anti-VEGF (bevacizumab) subconjuntival e tópica na neovascularização corneana experimental

Purpose: To compare the effects of bevacizumab applied either subconjunctivally or topically, in a rat model of corneal neovascularization induced by alkali burn. Methods: The right corneas of 24 Wistar-Albino rats were cauterized with silver nitrate sticks. The rats were divided randomly and equally into three groups: no treatment control (n=8), subconjunctival bevacizumab treatment (n=8), and topical bevacizumab treatment (n=8). Immediately following cauterization, the subconjunctival group was treated with a 0.05 ml (1.25 mg) bevacizumab subconjunctival injection. The topical group was treated with 10 mg/ml bevacizumab twice daily, and the control group received subconjunctival saline injections twice daily. The burn stimulus and neovascularization scores were evaluated using a technique previously described by Mahoney and Waterbury. Digital photographs were obtained before the eyes were enucleated and corneal sections were then analyzed by histopathology. Results: The mean burn stimulus score was 1.86 ± 0.6 and there was no statistical difference between the groups (p=0.730). The mean neovascularization scores in the subconjunctival and topical bevacizumab groups were statistically lower than the control group (p<0.05). The mean percentage area of corneal neovascularization was 82.5 ± 22.1 in the control group, 42.7 ± 15.0 in the subconjunctival group, and 55.8 ± 18.2 in the topical group. The differences between the control and treatment groups were statistically significant (p<0.05). Histopathology showed that the treatment groups presented less neovascularization, inflammation, and fibroblast activity than the control group (p<0.05). Conclusions: This study demonstrates that both subconjunctival and topical administrations of bevacizumab inhibit corneal neovascularization and decrease inflammation and fibroblast activity in a rat model of corneal neovascularization induced by alkali burn. .

Objetivo: Comparar o efeito de bevacizumab aplicado subconjuntival e topicamente em um modelo de neovascularização de córnea de ratos induzida por queimadura alcalina. Métodos: Córneas direitas de 24 ratos Wistar-Albino foram cauterizados por nitrato de prata. Os indivíduos foram divididos aleatoriamente e igualmente em três grupos: controle (n=8), o bevacizumab subconjuntival (n=8), o bevacizumab tópico (n=8). Imediatamente após a cauterização, 0,05 ml (1,25 mg) de bevacizumab foi injetado no grupo subconjuntival. Grupo tópico foi inculcado com 10 mg/ml de bevacizumab duas vezes por dia. O grupo controle recebeu solução salina normal, topicamente, duas vezes ao dia. A graduação do estímulo da queimadura e a graduação da neovascularização foram avaliados utilizando a técnica descrita por Mahoney e Waterbury. Fotografias digitais foram obtidas dos olhos serem enucleados. Seções da córnea foram analisadas por histopatologia. Resultados: A média da graduação do estímulo da queimadura foi de 1,86 ± 0,6 e não houve diferença estatisticamente entre os grupos (p=0,730). As médias das graduações da neovascularização no grupo bevacizumab subconjuntival e no grupo bevacizumab tópico foram estatisticamente menores do que o grupo controle (p<0,05). A percentagem média de área de neovascularização da córnea foi de 82,5 ± 22,1 no grupo controle, 42,7 ± 15,0 no grupo subconjuntival e 55,8 ± 18,2 no grupo tópico. As diferenças entre os grupos de tratamento e grupo de controlo foram estatisticamente significativos (p<0,05). A histopatologia mostrou que os grupos de tratamento apresentavam menos neovascularização, inflamação e atividade de fibroblastos do que o grupo controle (p<0,05). ...

An experimental model of chronic rhinosinusitis in rabbits without bacterial inoculation

Evaluate and compare two different experimental techniques of maxillary sinus ostium occlusion using N-butyl cyanoacrylate in developing chronic histological findings without the inoculation of pathogenic bacteria among rabbits. In a randomized study, sixteen New Zealand rabbits were assigned for occlusion of the right maxillary sinus through a transmaxillary approach or through the roof of the nasal cavity. The contralateral sinus served as a control. After 12 weeks, the animals were sacrificed for blinded histopathological analysis of the maxillary sinus mucosa. Histopathological changes consistent with CRS were found in eight (100%) of the maxillary sinuses approached transmaxillary and three of those through the roof of the nasal cavity (37.5%), p 0.008 and 0.250, respectively, comparing with the control side. Chronic mucosal changes were significantly better induced using the transmaxillary approach (p 0.026). It is possible to induce a model of chronic sinusitis among rabbits with transmaxillary sinus occlusion without bacterial inoculation. This model can be replicated for future cellular studies.(AU)

An experimental model of chronic rhinosinusitis in rabbits without bacterial inoculation

PURPOSE: Evaluate and compare two different experimental techniques of maxillary sinus ostium occlusion using N-butyl cyanoacrylate in developing chronic histological findings without the inoculation of pathogenic bacteria among rabbits. METHODS: In a randomized study, sixteen New Zealand rabbits were assigned for occlusion of the right maxillary sinus through a transmaxillary approach or through the roof of the nasal cavity. The contralateral sinus served as a control. After 12 weeks, the animals were sacrificed for blinded histopathological analysis of the maxillary sinus mucosa. RESULTS: Histopathological changes consistent with CRS were found in eight (100%) of the maxillary sinuses approached transmaxillary and three of those through the roof of the nasal cavity (37.5%), p 0.008 and 0.250, respectively, comparing with the control side. Chronic mucosal changes were significantly better induced using the transmaxillary approach (p 0.026). CONCLUSION: It is possible to induce a model of chronic sinusitis among rabbits with transmaxillary sinus occlusion without bacterial inoculation. This model can be replicated for future cellular studies. .

Median lethal needle caliber in two models of experimental sepsis

To estimate the median lethal needle caliber (LC50) of a new experimental sepsis model and compare it to the LC50 of the cecal ligation and puncture (CLP) sepsis model. METHODS: Male albino Wistar rats were studied (n=22). Animals were allocated into two study groups. In Group I, experimental sepsis was induced by cecal ligation and puncture. In Group II, experimental sepsis was induced by ascending colon ligation and cecal puncture. Up-and-down method was used to determinate the LC50. RESULTS: LC50 in Group I was 19 Gauge (Confidence Interval 17 to 22 Gauge). Determination of LC50 was not possible in Group II due to the death of all animals. CONCLUSION: LC50 in cecal ligation and puncture is 19 Gauge. The lethality of the new model tested in this trial is very high.(AU)

Median lethal needle caliber in two models of experimental sepsis

To estimate the median lethal needle caliber (LC50) of a new experimental sepsis model and compare it to the LC50 of the cecal ligation and puncture (CLP) sepsis model. METHODS: Male albino Wistar rats were studied (n=22). Animals were allocated into two study groups. In Group I, experimental sepsis was induced by cecal ligation and puncture. In Group II, experimental sepsis was induced by ascending colon ligation and cecal puncture. Up-and-down method was used to determinate the LC50. RESULTS: LC50 in Group I was 19 Gauge (Confidence Interval 17 to 22 Gauge). Determination of LC50 was not possible in Group II due to the death of all animals. CONCLUSION: LC50 in cecal ligation and puncture is 19 Gauge. The lethality of the new model tested in this trial is very high.

A new experimental model of cigarette smoke-induced emphysema in Wistar rats / Um novo modelo experimental murino de enfisema: enfisema induzido pela fumaça do cigarro em ratos Wistar

OBJECTIVE: To describe a new murine model of cigarette smoke-induced emphysema. METHODS: Twenty-four male Wistar rats were divided into two groups: the cigarette smoke group, comprising 12 rats exposed to smoke from 12 commercial filter cigarettes three times a day (a total of 36 cigarettes per day) every day for 30 weeks; and the control group, comprising 12 rats exposed to room air three times a day every day for 30 weeks. Lung function was assessed by mechanical ventilation, and emphysema was morphometrically assessed by measurement of the mean linear intercept (Lm). RESULTS: The mean weight gain was significantly (approximately ten times) lower in the cigarette smoke group than in the control group. The Lm was 25.0% higher in the cigarette smoke group. There was a trend toward worsening of lung function parameters in the cigarette smoke group. CONCLUSIONS: The new murine model of cigarette smoke-induced emphysema and the methodology employed in the present study are effective and reproducible, representing a promising and economically viable option for use in studies investigating the pathophysiology of and therapeutic approaches to COPD. .

OBJETIVO: Descrever um novo modelo murino de enfisema induzido pela fumaça de cigarro. MÉTODOS: Vinte e quatro ratos Wistar foram divididos em dois grupos: o grupo fumaça de cigarro, com 12 ratos expostos à fumaça de 12 cigarros comerciais com filtro três vezes ao dia (um total de 36 cigarros por dia), sete dias por semana, durante 30 semanas e o grupo controle, com 12 animais expostos ao ar ambiente três vezes ao dia, sete dias por semana, durante 30 semanas. A função pulmonar foi avaliada por meio de ventilação mecânica, e o enfisema foi morfometricamente avaliado por meio do diâmetro alveolar médio (Lm). RESULTADOS: A média de ganho de peso foi significativamente menor (aproximadamente dez vezes menor) no grupo fumaça de cigarro do que no grupo controle. O Lm foi 25.0% maior no grupo fumaça de cigarro. Os parâmetros de função pulmonar tenderam a ser piores no grupo fumaça de cigarro. CONCLUSÕES: O novo modelo murino de enfisema induzido pela fumaça de cigarro e a metodologia empregada neste estudo são eficazes e reproduzíveis; são, portanto, uma opção promissora e economicamente viável para estudos sobre a fisiopatologia e o tratamento da DPOC. .

A new experimental model for inducing interstitial cystitis by oxidative stress using intravesical instillation of a nitric oxide donor gel / Novo modelo experimental de indução de cistite intersticial por estresse oxidativo utilizando instilação intravesical de gel doador de óxido nítrico

Objective: The aim of this study was to develop an experimental model of inducing interstitial cystitis through intravesical instillation of a polymeric solution containing the NO donor S-nitrousglutathione (GSNO) and to compare it to the experimental interstitial cystitis induced by vesical instillation of protamine and potassium chloride. Methods: A total of 40 female Wistar rats were used and divided into four groups: 1 - ten rats treated with saline solution + GSNO; 2 - ten rats treated with saline solution + polymeric solution (without GNSO); 3 - ten rats treated with protamine sulphate + KCl; 4 - ten rats treated with protamine sulphate + GSNO. The rats received one application (five animals in each group) or three applications (five animals in each group) of the corresponding substance through intravesical instillation, and after six days (five animals in each group) or nine days (five animals in each group) they were euthanized and their bladders were removed for macroscopic evaluation and histological study. Results: In the macroscopic evaluation edema and hyperemia of the mucosa were observed in 2 (22%) animals in Group 1, in no (0%) animal in Group 2, in 10 (100%) animals in Group 3, and in 5 (50%) animals in Group 4. In the protamine + KCl group and in saline + GSNO, similar effects were observed in the bladder wall. The animals in Group 2 (saline + polymeric solution) showed significantly less vascular congestion compared to the other groups after 9 days of the instillation (p = 0.0035). Significant fibrosis was observed in Groups 3 and 4, 6 days (p = 0.3781) and 9 days (p = 0.0459) after instillations, when compared to controls (Group 2). All groups presented neutrophilic infiltrate of variable intensity, 6 days after instillations (p = 0.7277). After 9 days, there was a regression of the infiltrate, with no evidence of accentuated neutrophilic reaction in all the groups (p = 0.2301). Conclusions: The inflammatory response to bladder instillation with an aqueous solution of S-nitrousglutathione was very similar to that induced by bladder instillation of protamine and KCl. Instillation of an aqueous solution of S-nitrousglutathione can be considered a new model for experimental induction of interstitial cystitis.

Objetivo: O objetivo deste estudo foi o desenvolvimento de um modelo experimental para a indução de cistite intersticial, por meio da instilação vesical de uma solução polimérica de gel doador de óxido nítrico S-nitrosoglutationa (GSNO), e compará-lo ao modelo experimental para a indução da cistite intersticial por instilação vesical de protamina e cloreto de potássio. Métodos: Foram utilizadas 40 ratas Wistar, divididas em quatro grupos: 1 - dez ratas tratadas com solução salina + GSNO; 2 - dez ratas tratadas com solução salina + solução de polímeros (sem GNSO); 3 - dez ratas tratadas com sulfato de protamina + KCl; 4 - dez ratas tratadas com sulfato de protamina + GSNO. As ratas receberam uma aplicação (cinco animais em cada grupo) ou três aplicações (cinco animais em cada grupo) da substância correspondente através de instilação vesical, e após seis dias (cinco animais em cada grupo) ou nove dias (cinco animais em cada grupo) foram sacrificadas, e a bexiga foi removida para exame macroscópico e estudo histológico. Resultados: Na avaliação macroscópica observou-se edema e hiperemia da mucosa em 2 animais (22%) do Grupo 1, em nenhum animal (0%) do Grupo 2, em 10 animais (100%) do Grupo 3, e em 5 animais (50%) do Grupo 4. No grupo protamina + KCl e no grupo solução salina + GSNO, observamos efeitos semelhantes sobre a parede da bexiga. Os animais do Grupo 2 (salina + polímeros) apresentaram significantemente menos congestão vascular que os dos outros grupos após 9 dias de instilação (p = 0,0035). Observou-se fibrose significante nos Grupos 3 e 4, 6 dias (p = 0,3781) e 9 dias (p = 0,0459) após as instilações, quando comparados com o grupo controle (Grupo 2). Todos os grupos apresentaram infiltrados neutrofílicos de intensidade variável, 6 dias após as instilações (p = 0,7277). Após 9 dias, observou-se regressão do infiltrado, sem evidência de acentuada reação neutrofílica em todos os grupos (p = 0,2301). Conclusão: A resposta inflamatória à instilação da bexiga com uma solução aquosa de S-nitroglutationa foi muito semelhante àquela induzida pela instilação de protamina e KCl. A instilação de uma solução aquosa de S-nitroglutationa pode ser considerada um novo modelo experimental para a indução da cistite intersticial.

Modelo experimental para restrição do crescimento fetal em ratos: efeito sobre o glicogênio hepático e morfometria intestinal e renal / Experimental rat model for fetal growth restriction: effects on liver glycogen and intestinal and renal morphometry

OBJETIVO: avaliar a eficácia do modelo de RCIU por ligadura da artéria uterina simulando insuficiência placentária em ratos. MÉTODOS: fetos de ratas prenhes Sprague-Dawley foram divididos em três grupos: RCIU (restrição de crescimento intrauterino), com fetos submetidos à ligadura da artéria uterina com 18,5 dias de gestação (termo = 22 dias), C-RCIU (controle da restrição), com fetos do corno contralateral à ligadura, CE (Controle Externo), com fetos de ratas sem manipulação. Com 21,5 dias de gestação, foi realizada cesárea, os fetos foram pesados e dissecados para análise morfométrica e histológica do fígado, intestino e rins. RESULTADOS: os dados morfométricos avaliados mostraram o peso corpóreo (PC), hepático (PH) e intestinal (PI) dos fetos com RCIU menor que C-RCIU e CE (p<0,001). O peso placentário (PP), renal (PR) e as relações PH/PC, PI/PC e PR/PC não se alteraram. A espessura renal foi menor nos fetos com RCIU (p<0,001) e houve diminuição da camada mucosa e submucosa intestinal (p<0,05). A avaliação histológica mostrou diminuição do glicogênio hepático nos fetos com RCIU em relação aos grupos C-RCIU e CE. CONCLUSÕES: o modelo descrito foi eficiente e causou RCIU fetal simétrica com diminuição da maioria dos órgãos, especialmente do peso hepático, e alteração nos depósitos de glicogênio.

PURPOSE: to evaluate the effectiveness of the IUGR model by uterine artery ligation mimicking placental insufficiency in rats. METHODS: sprague-Dawley rat fetuses were divided into three groups: IUGR (intrauterine growth restriction), with fetuses in the right horn of pregnant rats subjected to right uterine artery ligation at 18.5 days of gestation (term = 22 days); C-IUGR (control of restriction), with control fetuses in the left horn, and EC (external control), with fetuses of intact rats. Animals were harvested by cesarean section at day 21.5 days of gestation. Fetuses were weighed and then sacrificed. The intestine, liver, kidney and placenta were weighed and dissected for morphometric and histological analysis. RESULTS: the morphometric data showed decreased body weight (BW), liver weight (LW) and intestinal weight (IW) of fetuses with IUGR compared to C-IUGR and EC (p<0.001). The placental weight (PW), renal weight (RW) and LW/BW, IW/BW, and RW/BW ratios did not change. IUGR fetuses had decreased kidney thickness (p<0.001) and decreased thickness of the intestinal mucosa and submucosa (p<0.05). Histological evaluation showed reduction of liver glycogen storage in fetuses with IUGR compared to C-IUGR and CE. CONCLUSIONS: the model described was efficient and caused symmetric fetal IUGR with decreased size of most organs, especially the liver, and changes in glycogen stores.