Reduction of oxidative stress improves insulin signaling in cardiac tissue of obese mice / Redução do estresse oxidativo melhora a sinalização da insulina em tecido cardíaco de camundongos obesos

ABSTRACT Objective To evaluate the effects of oxidative stress on insulin signaling in cardiac tissue of obese mice. Methods Thirty Swiss mice were equally divided (n=10) into three groups: Control Group, Obese Group, and Obese Group Treated with N-acetylcysteine. After obesity and insulin resistance were established, the obese mice were treated with N-acetylcysteine at a dose of 50mg/kg daily for 15 days via oral gavage. Results Higher blood glucose levels and nitrite and carbonyl contents, and lower protein levels of glutathione peroxidase and phosphorylated protein kinase B were observed in the obese group when compared with their respective control. On the other hand, treatment with N-acetylcysteine was effective in reducing blood glucose levels and nitrite and carbonyl contents, and significantly increased protein levels of glutathione peroxidase and phosphorylated protein kinase B compared to the Obese Group. Conclusion Obesity and/or a high-lipid diet may result in oxidative stress and insulin resistance in the heart tissue of obese mice, and the use of N-acetylcysteine as a methodological and therapeutic strategy suggested there is a relation between them.

RESUMO Objetivo Avaliar os efeitos do estresse oxidativo sobre a sinalização da insulina em tecido cardíaco de camundongos obesos. Métodos Utilizaram-se 30 camundongos Swiss subdivididos igualmente (n=10) em três grupos: Grupo Controle, Grupo Obeso e Grupo Obeso Tratado com N-acetilcisteína. Após estabelecidas a obesidade e a resistência à insulina, os camundongos obesos foram tratados diariamente, durante 15 dias, via gavagem oral, com N-acetilcisteína na dose de 50mg/kg. Resultados Observaram-se maiores níveis de glicose sanguínea, conteúdos de nitrito e carbonil, e menores níveis proteicos de glutationa peroxidase e proteína quinase B fosforilada no Grupo Obeso quando comparado a seu respectivo controle. Por outro lado, o tratamento com N-acetilcisteína se mostrou eficiente em diminuir os níveis glicêmicos, os conteúdos de nitrito e carbonil, e aumentar significativamente os níveis proteicos de glutationa peroxidase e proteína quinase B fosforilada, quando comparados ao Grupo Obeso. Conclusão Obesidade e/ou dieta hiperlipídica levam a estresse oxidativo e à resistência à insulina no tecido cardíaco de camundongos obesos, e o uso da N-acetilcisteína como estratégia metodológica e terapêutica sugeriu haver relação entre ambos.

Losartan improves alveolar bone dynamics in normotensive rats but not in hypertensive rats

Abstract Hypertension is one of the main causes of premature death in the world; also, it is associated with several bone alterations. Preclinical studies have demonstrated delayed alveolar bone healing in hypertensive rats. However, losartan has been favorable for consolidation of bone grafts and reduction in active periodontitis. Therefore, losartan is suggested to be effective in bone formation stages, as well as in the synthesis of matrix proteins and mineralization. Objectives: To evaluate the alveolar bone dynamics in hypertensive rats treated with losartan by laser confocal microscopy and histological analysis. Methodology: Thirty-two rats, 16 spontaneously hypertensive rats (SHR) and 16 Wistar albinus rats, treated or not with losartan (30 mg/kg/day) were used. Calcein fluorochrome at 21 days and alizarin red fluorochrome at 49 days were injected in rats (both 20 mg/kg). The animals were submitted to euthanasia 67 days after treatment, and then the right maxilla was removed for laser confocal microscopy analysis and the left maxilla for histological analysis. Results: This study showed a greater calcium marking in normotensive animals treated with losartan in relation to the other groups. Laser confocal microscopy parameters showed higher values of bone volume formed, mineralized surface, active surface of mineralization and bone formation rate in normotensive animals treated with losartan. However, a smaller mineralized surface was observed in all hypertensive animals. Conclusion: Losartan can improve bone mineralization parameters under normal physiological conditions, but the same anabolic effect does not occur under hypertension.

Cryotherapy: biochemical alterations involved in reduction of damage induced by exhaustive exercise

When exercises are done in intense or exhaustive modes, several acute biochemical mechanisms are triggered. The use of cryotherapy as cold-water immersion is largely used to accelerate the process of muscular recovery based on its anti-inflammatory and analgesic properties. The present study aimed to study the biochemical effects of cold-water immersion treatment in mice submitted to exercise-induced exhaustion. Swiss albino mice were divided into 4 treatment groups: control, cold-water immersion (CWI), swimming exhaustive protocol (SEP), and SEP+CWI. Treatment groups were subdivided into times of analysis: 0, 1, 3, and 5 days. Exhaustion groups were submitted to one SEP session, and the CWI groups submitted to one immersion session (12 min at 12°C) every 24 h. Reactive species production, inflammatory, cell viability, and antioxidant status were assessed. The SEP+CWI group showed a decrease in inflammatory damage biomarkers, and reactive species production, and presented increased cell viability compared to the SEP group. Furthermore, CWI increased acetylcholinesterase activity in the first two sessions. The present study showed that CWI was an effective treatment after exercise-induced muscle damage. It enhanced anti-inflammatory response, decreased reactive species production, increased cell viability, and promoted redox balance, which could decrease the time for the recovery process.

Endoscopic repair of cerebrospinal fluid rhinorrhea / Reparo endoscópico de fístulas liquóricas nasais

Abstract Introduction: Nasal liquorrhea indicates a cerebrospinal fluid fistula, an open communication between the intracranial cerebrospinal fluid and the nasal cavity. It can be traumatic and spontaneous. Objective: The aim of this study was to assess the outcome of endoscopic repair of cerebrospinal fluid fistula using fluorescein. Methods: This retrospective study included 30 patients of both sexes, with a mean age of 48.7 years, treated in the period from 2007 to 2015. All patients underwent lumbar administration of 5% sodium fluorescein solution preoperatively. Fistula was closed using three-layer graft and fibrin glue. Results: Cerebrospinal fluid fistulas were commonly located in the ethmoid (37%) and sphenoid sinus (33%). Most patients presented with traumatic cerebrospinal fluid fistulas (2/3 of patients). The reported success rate for the first repair attempt was 97%. Complications occurred in three patients: one patient presented with acute hydrocephalus, one with reversible encephalopathy syndrome on the fifth postoperative day with bilateral loss of vision, and one patient was diagnosed with hydrocephalus two years after the repair of cerebrospinal fluid fistula. Conclusion: Endoscopic diagnosis and repair of cerebrospinal fluid fistulas using fluorescein intrathecally has high success rate and low complication rate.

Resumo Introdução: A liquorreia nasal indica uma fístula liquórica (FL), uma comunicação aberta entre o líquido cerebrospinal intracraniano e a cavidade nasal. Pode ser traumática e espontânea. Objetivo: Avaliar o desfecho do reparo endoscópico da fístula liquórica nasal com o uso de fluoresceína. Método: Este estudo retrospectivo incluiu 30 pacientes de ambos os sexos, com idade média de 48,7 anos, tratados de 2007 a 2015. Todos os pacientes foram submetidos à administração lombar de solução de fluoresceína de sódio a 5% no pré-operatório. A fístula foi fechada com enxerto de três camadas e cola de fibrina. Resultados: As fístulas de líquido cerebrospinal foram comumente localizadas no seio etmoidal (37%) e esfenoidal (33%). A maioria dos pacientes apresentou fístulas liquóricas traumáticas (2/3). A taxa de sucesso relatada para a primeira tentativa de reparo foi de 97%. Complicações ocorreram em três pacientes: um apresentou hidrocefalia aguda; um, síndrome reversível de encefalopatia no quinto dia de pós-operatório com perda bilateral da visão; e um foi diagnosticado com hidrocefalia dois anos após o reparo de fístula liquórica. Conclusão: O diagnóstico endoscópico e o reparo de fístulas liquóricas nasais com uso de fluoresceína intratecal têm alta taxa de sucesso e baixo índice de complicações.

Obstrução congênita de vias lacrimais: série de casos de sondagem e intubação / Congenital lacrimal obstruction: case series of probing and intubation

RESUMO Objetivo: Avaliar a relação entre a idade da intervenção cirúrgica em pacientes portadores de obstrução congênita das vias lacrimais e o sucesso obtido pelos mesmos. Métodos: Foram estudados 94 prontuários de pacientes com diagnóstico de obstrução congênita de vias lacrimais, assistidos no setor de oculoplástica do Hospital de Olhos do Paraná, no período de janeiro de 2006 a dezembro de 2012. Para diagnóstico e inclusão na pesquisa foram realizados a anamnese, exames oftalmológico geral e específico utilizando o Teste de Milder e o Teste de Obtenção de Fluoresceína na Orofaringe. Para determinar o tipo de intervenção (sondagem ou intubação) foram utilizados os critérios do serviço. Na análise estatística foram utilizados o teste de qui-quadrado e o teste t de Student e adotado o nível de significância de 5%. Resultados: Quando comparados os resultados por idade, independente do tratamento, não houve associação significativa (p=0,223) entre a taxa de melhora e a idade. Conclusão: No presente estudo não houve diferença significativa entre a intervenção cirúrgica e as faixas etárias abordadas.

ABSTRACT Objective: To evaluate the relationship between age at surgery in patients with congenital lacrimal obstruction and success achieved by them. Methods: We studied 94 medical records of patients diagnosed with congenital lacrimal obstructions, assisted at the oculoplastic sector of the Hospital de Olhos do Paraná, in the period january 2006 to december 2012. For diagnosis and inclusion in the study, the clinical history were made, overall and specific ophthalmologic examination, with the tests: Milder and Observation Test of Fluorescein in the Oropharynx. To determine the type of intervention (probing or intubation) service criteria were used. In the statistical analysis, the chi-square and Student’s t test were used. Significance level of 5 % was adopted. Results: When comparing the results by age, regardless of treatment, there was no significant association (p = 0.223) between the rate of improvement and age. Conclusions: In the present study shows, there is no significant difference between the surgical interventions within the age groups addressed.

Chemical and cytological analysis of cerebral spinal fluid after intrathecal injection of hypodense fluorescein / Análise química e citológica do líquor após injeção intratecal de solução hipodensa de fluoresceína

ABSTRACT INTRODUCTION: Intrathecal fluorescein has been effective for topographic diagnosis of rhinoliquorrhea. Nonetheless, there are no reports on the study of cerebral spinal fluid (CSF) after use of intrathecal fluorescein. OBJECTIVE: A prospective study attempting to evaluate CSF through chemical and cytological analysis, after injection of fluorescein. METHODS: Prospective analysis of 24 samples of CSF after intrathecal injection of fluorescein for topographic diagnosis of CSF fistulae, collected at the time of puncture and after 24 and 48 h, divided by cellularity: Group 1, up to five cells, and Group 2, with more than five cells. RESULTS: The yellow-greenish color of CSF remained after 48 h in 36%, evidencing permanence of fluorescein. No changes in protein and glucose levels were observed between 0-24 h and 0-48 h. In group 2, an increase in cell count was observed between 24 h and 48 h (p = 0.019). In both groups, there was an increase of neutrophils between 0 and 48 h (p = 0.048) and a decrease between 24 and 48 h (p = 0.05). CONCLUSION: Intrathecal fluorescein provoked discreet meningeal reactions, such as an increase of cells between 24 and 48 h and an increase of neutrophils at 24 h, with a subsequent decrease at 48 h with no correlation with symptomatology.

RESUMO Introdução: A fluoresceína intratecal tem sido efetiva no diagnóstico topográfico da rinoliquorréia. Entretanto, não há estudos no líquor após o uso de fluoresceína intratecal. Objetivo: Estudo prospectivo visando avaliar o líquor, através de análise química e citológica, após injeção de fluoresceína. Método: Análise prospectiva de 24 punções após injeção intratecal de fluoresceína para diagnóstico topográfico de fístula liquórica, coletado no momento da punção, 24 e 48 horas, divididos pela celularidade: grupo 1, com até 5 células e grupo 2 com mais de 5 células. Resultado: A coloração amarelo-esverdeada do líquor permaneceu após 48 horas em 36%, evidenciando permanência de fluoresceína. Observou-se ausência de mudanças no nível de proteína e glicose entre 0-24 horas e 0-48 horas. No grupo 2, um aumento na contagem celular foi observado entre 24 e 48 horas (p = 0,019). No dois grupos juntos, observou-se um aumento de neutrófilos entre 0 e 48 horas (p = 0,048) e uma diminuição entre 24 e 28 horas (p = 0,05). Conclusão: Fluoresceína intratecal provocou discretas reações meníngeas, como o aumento de células entre 24 e 48 horas e aumento dos dos neutrófilos em 24 horas com uma subsequente dimi nuição em 48 horas sem correlação com sintomas.

Resección microquirúrgica de glioblastoma guiada con fluoresceína intraoperatoria: evaluación retrospectiva / Microsurgical resection of glioblastoma guided with intraoperative fluorescein: a retrospective evaluation

Evaluar la influencia del uso de fluoresceína sódica (FLS-Na) en la cirugía del glioblastoma (GB) sobre el grado de resección tumoral y la supervivencia en pacientes atendidos en el Instituto Nacional de Enfermedades Neoplásicas. Materiales y métodos. Se revisó un total de 238 casos de GB atendidos entre los años 2008 y 2013 y se seleccionó 150 casos de GB sometidos a resección quirúrgica, con información clínico-patológica y seguimiento adecuado. Resultados. La media de edad fue 51 años, el 58,7% de casos presento Karnofsky de al menos 90. Se administró FLS-Na en 80 casos (53,3%) y se obtuvo una resección subtotal y total en 69 (46%) y 81 (54%) de los casos, respectivamente. El grupo que recibió FLS-Na obtuvo mayores tasas de resección total que el grupo operado solo con luz blanca (77,5 vs 27,1%, p<0,001). La mediana de sobrevida global (SG) fue mayor en el grupo sometido a resección total que a subtotal (17 vs 7 meses, p<0,001). La mediana de SG en los que recibieron FLS-Na fue mayor que en los que no la recibieron (15,0 vs 8 meses, p=0,003). Otros factores que afectaron la SG fueron la edad (p=0,002), el Karnofsky (p=0,052) y la administración de radioterapia (p=0,016) y quimioterapia (p=0,011). Conclusiones. La técnica microquirúrgica con administración de FLS-Na se asoció con un aumento en la tasa de resecciones totales y de supervivencia...

To evaluate the influence of the use of sodium fluorescein (FLS-Na) in surgery of glioblastoma (GB) on the degree of tumor resection and survival in patients treated at the National Institute of Neoplastic Diseases. Materials and methods. A total of 238 cases of GB treated between 2008 and 2013 were reviewed and 150 cases of GB who underwent surgical resection with clinicopathological information and adequate follow-up were selected. Results. The mean age was 51 years, 58.7% of the cases presented a Karnofsky score of at least 90. FLS-Na was administered in 80 cases (53.3%) and a subtotal and total resection was obtained in 69 (46%) and 81 (54%) cases, respectively. The group that received FLS-Na obtained higher rates of total resection than the group operated with white light alone (77.5 vs 27.1%, p<0.001). The median overall survival (OS) was higher in the group subject to total compared to subtotal resection (17 vs 7 months, p<0.001). The median OS in those who received FLS-Na was higher than in those who did not (15.0 vs 8 months, p=0.003). Other factors affecting OS were age (p=0.002), the Karnofsky score (p=0.052) and radiation therapy (p=0.016) and chemotherapy (p=0.011). Conclusions. The microsurgical technique with administration of FLS-Na was associated with an increase in the rate of total resection and survival...

Síndrome de Fraser: relato de caso nas vias lacrimais / Fraser Syndrome: case report in lacrimal system

A síndrome de Fraser é uma condição sistêmica caracterizada por criptoftalmo, sindactilia e anomalia da genitália, podendo se associar com alterações dos rins, do ouvido, do nariz, da laringe e do esqueleto. O criptoftalmo pode representar um achado isolado, representado por herança autossômica dominante, associado a outras anomalias congênitas, relatado como herança autossômica recessiva. Criança do sexo feminino, 9 meses, avaliada no ambulatório de vias lacrimais da Universidade Federal de São Paulo. Filha de pais consanguíneos. Ao exame, foram observados criptoftalmo total à esquerda, epífora em olho direito associada à secreção mucopurulenta, nariz em sela, implantação baixa das orelhas, malformação de conduto auditivo, aumento de grandes lábios e sindactilia de mãos e pés. A tomografia de crânio evidenciou braquicefalia ausência de septo pelúcido, proeminência dos ventrículos laterais, importante falha óssea na calota craniana, presença de afilamento do manto tecidual cerebral, fossa posterior pequena, desorganização do segmento anterior, afacia e descolamento total da retina.

Fraser syndrome is a systemic condition characterized by cryptophthalmos, syndactyly and abnormal genitalia, which may be associated with urinary tract, ear, nose, larynx and skeletal abnormalities. Cryptophthalmos can be an isolated finding (that has been reported as an autosomal dominant trait) or associated with other congenital anomalies (reported as an autosomal recessive disorder). Child, female, nine month of life, evaluated in the lacrimal setor of Federal University of São Paulo. Child of consanguineous parents. Her physical examination showed total unilateral cryptophthalmos (left side), epiphora (right side) with mucopurulent discharge, depressed nasal bridge, low set ears, atresia of the external auditory canal, prominent labia majora and syndactyly of the fingers and toes. Ocular ultrasonography showed brachycephaly, absence of septu pellucidum prominence of the lateral ventricles, a major bone defect in the skull, the presence of thinning of the mantle tissue of the brain,a reduced anterior-posterior ocular diameter, anterior segment disorganization, absence of the lens and total retinal detachment in the left eye.

Análisis comparativo de marcadores de lesión en modelos de isquemia cerebral focal y global en ratas / Injury markers in two models of cerebral ischemia

Introducción. Los indicadores espacio-temporales de lesión son esenciales en el estudio neuropatológico y terapéutico de la isquemia cerebral. Objetivo. Optimizar la técnica de dos modelos de isquemia cerebral (focal y global) y hacer un análisis comparativo de la progresión del daño cerebral, mediante marcadores de neurodegeneración. Materiales y métodos. Se sometieron ratas Wistar a oclusión temporal de la arteria cerebral media o a oclusión de cuatro vasos, y se evaluaron comparativamente el tiempo quirúrgico, la tasa de supervivencia y la recuperación neurológica. Se utilizó trifenilo de tetrazolio para establecer la distribución del infarto y tinción con Fluoro - Jade B ® como marcador de neurodegeneración. La inmunorreacción de la astroglía se evaluó con el anticuerpo contra la proteína acídica fibrilar de la glía ( Glial Fibrillary Acidic Protein, GFAP) y el anticuerpo AT-8 contra la proteína tau hiperfosforilada, 24, 48 y 72 horas después de la isquemia. Resultados. Los modelos de isquemia utilizados requirieron menor tiempo quirúrgico y hubo menor riesgo de muerte, respecto a estudios previos. En el modelo focal, las células positivas con Fluoro - Jade B ® y los astrocitos reactivos, se evidenciaron en corteza e hipocampo a las 24 horas después de la isquemia. En el modelo global, se observó tinción Fluoro - Jade B ® positiva a las 24 horas, aumentando significativamente la reacción de la GFAP a las 72 horas en corteza y a las 48 horas en el hipocampo. La reacción contra la proteína tau hiperfosforilada aumentó progresivamente y fue máxima a las 72 horas en ambos modelos. Conclusiones. Los dos modelos de isquemia cerebral, oclusión temporal de la arteria cerebral media y oclusión de cuatro vasos, fueron optimizados. En estos modelos, los marcadores la tinción Fluoro - Jade B ® y la GFAP permitieron detectar procesos de neurodegeneración 24 horas después de la isquemia, en tanto el marcador de proteína tau hiperfosforilada (AT-8) incrementó progresivamente su reacción hasta las 72 horas, lo cual sugiere la propagación de la excitotoxicidad y la alteración de enzimas implicadas en la fosforilación de proteínas del citoesqueleto.

Introduction: Spatio-temporal indicators of injury are essential for the study of neuropathological processes and for developing therapeutic approaches for stroke. Objective: This study sought to optimize the techniques of two cerebral ischemia models (focal and global) and to comparatively evaluate the progression of brain damage by analyzing markers of neurodegeneration. Materials and methods: Wistar rats were subjected to temporary occlusion of the middle cerebral artery (t-MCAO) or four-vessel occlusion (4-VO), and surgical time, survival rate and neurological recovery were comparatively evaluated. Triphenyl tetrazolium was used to determine the distribution of the infarction, and Fluoro-Jade B was used as a marker of neurodegeneration. Astroglial immunoreactivity was assessed with an anti-glial fibrillary acidic protein (GFAP) antibody, and an anti-AT-8 antibody was used to detect hyperphosphorylated tau protein at 24, 48 and 72 hours post-ischemia. Results: The cerebral ischemia models employed (t-MCAO and 4-VO) required less surgical time and presented less of a death risk compared to those in previous studies. In the focal model, Fluoro-Jadepositive cells and reactive astrocytes were observed in the cerebral cortex and the hippocampus at 24 hours post-ischemia. In the global model, we observed Fluoro-Jade-positive cells at 24 hours, and a significant increase in the reactivity of GFAP was observed at 72 hours in the cortex and at 48 hours in the hippocampus. The immunoreactivity of hyperphosphorylated tau protein increased progressively, reaching a maximum at 72 hours post-ischemia in both models. Conclusions: These results suggest that in the t-MCAO and 4-VO ischemia models, the expression of Fluoro-Jade and GFAP indicates early neurodegeneration at 24 hours post-insult. In contrast, the immunoreactivity of the hyperphosphorylated tau protein marker (AT-8) progressively increases until 72 hours post-insult, which suggests that the progression of excitotoxicity and alteration of enzymes involves the phosphorylation of cytoskeletal proteins.

Anafilaxia com convulsões após angiografia com fluoresceína em paciente ambulatorial / Anaphylaxis with convulsions following intravenous fluorescein angiography at an outpatient clinic

A fluoresceína endovenosa é associada a pequenos efeitos adversos durante a angiografia da retina, a maioria deles sem gravidade e não relacionada a mecanismos imunológicos. Entretanto, em raras ocasiões, pode ocorrer reação anafilactoide; fatalidades envolvendo o uso de fluoresceína têm sido descritas. Complicações graves após injeção endovenosa de fluoresceína requerem ação imediata da equipe envolvida, assim como treinamento regular para padronizar o tratamento.

Fluorescein is associated with minor adverse effects during retinal angiography, and most of these effects are not severe and not related to immunological mechanism. However, on rare occasions, anaphylatoid reaction can occur, and fatalities involving fluorescein have been described. Life threatening complications after intravenous injection of sodium fluorescein require immediate intervention. Trainings for professionals are needed to standardize treatment for this event.

Bone regeneration in surgically created defects filled with autogenous bone: an epifluorescence microscopy analysis in rats

Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. OBJECTIVES: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. MATERIAL AND METHODS: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. RESULTS: The mineralization process was more intense in the graft group (32.09 percent) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66 percent). CONCLUSIONS: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

Niveles séricos de tetrayodotironina libre (T4L), mediante el método de electroquimioluminiscencia en caninos / Free tetraiodothyronine (FT4) serum levels by electrochemiluminescence method in canines

El presente estudio establece valores de referencia para niveles séricos de tetrayodotironina libre (T4L) en caninos mediante el método de electroquimioluminiscencia. Se utilizaron 180 caninos que fueron divididos en grupos según la edad y el sexo. Se encontraron diferencias altamente significativas (P<0,0001) relacionadas con la edad, sin encontrarse diferencias significativas con respecto al sexo para dicha hormona. Los resultados de este estudio sugieren que las concentraciones séricas de tetrayodotironina libre (ng/L) en caninos menores de 1año, de 1 a 7 años y mayores de 7 años, oscilan entre 9,90-11,74 ng/L, 8,51-11,74 ng/L y 7,48-8,64 ng/L, respectivamente. La determinación de T4L mediante electroquimioluminiscencia, puede considerarse útil como ayuda diagnóstica de posibles alteraciones tiroideas.

The present study establishes references values for free Tetraiodotironine (FT4) in canines using eletrochemiluminescence method. Blood samples from 180 canines divided in six groups of age (males and females), 30 animals for each group were used. Significant differences (P<0.0001) was found between age groups but not between sex groups. The canine average values for FT4 using this technique were as follow: younger than 1 year of age , 9.9 - 11.7 ng/L; from 1 to 7 years of age, 8.1 - 11.7 ng/L; older than 7 years of age 7.4- 8.6 ng/L. The electrochemiluminiscence method for measuring FT4 is valuable diagnostic tool in canine medicine.

Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection

Evaluation of apoptosis by flow cytometry is generally accomplished by methods that use annexin V-FITC as vital dye, which access phosphatidylserine exposed on the external membrane at the beginning of this process. In addition, the concomitant use of propidium iodide makes possible to verify the characteristic nuclear alterations in the late stages of apoptosis, as a consequence of the increase in membrane permeability. On the other hand, the use of calcein-AM in association with ethidium homodimer (EthD-1) allows the evaluation of cell apoptosis through detection of esterase activity and cellular membrane physical and chemical alterations. The aim of this study was to compare the sensibility of calcein-AM and EthD-1 with annexin V-FITC and propidium iodide for early apoptosis evaluation in peripheral blood mononuclear cell culture, obtained from HIV-infected patients. Apoptosis and cellular viability were detected and quantified by flow cytometry after 24 and 48 hours incubation times. Our results showed that calcein-AM/EthD-1 was more sensitive for apoptotic cell quantification in both incubation times than annexin V-FITC/propidium iodide (mean of 46.95 percent ± 3.56, p < 0.0001, for 24 hours and mean of 37.67 percent ± 2.47, p < 0.0014 for 48 hours), besides allowing to clearly define viable, apoptotic and dead cell populations.

Cl- and regulation of pH by MDCK-C11 cells

The interaction between H+ extrusion via H+-ATPase and Cl- conductance was studied in the C11 clone of MDCK cells, akin to the intercalated cells of the collecting duct. Cell pH (pHi) was measured by fluorescence microscopy using the fluorescein-derived probe BCECF-AM. Control recovery rate measured after a 20 mM NH4Cl acid pulse was 0.136 ± 0.008 pH units/min (dpHi/dt) in Na+ Ringer and 0.032 ± 0.003 in the absence of Na+ (0 Na+). With 0 Na+ plus the Cl- channel inhibitor NPPB (10 æM), recovery was reduced to 0.014 ± 0.001 dpHi/dt. 8-Br-cAMP, known to activate CFTR Cl- channels, increased dpHi/dt in 0 Na+ to 0.061 ± 0.009 and also in the presence of 46 nM concanamycin and 50 æM Schering 28080. Since it is thought that the Cl- dependence of H+-ATPase might be due to its electrogenic nature and the establishment of a +PD (potential difference) across the cell membrane, the effect of 10 æM valinomycin at high (100 mM) K+ was tested in our cells. In Na+ Ringer, dpHi/dt was increased, but no effect was detected in 0 Na+ Ringer in the presence of NPPB, indicating that in intact C11 cells the effect of blocking Cl- channels on dpHi/dt was not due to an adverse electrical gradient. The effect of 100 æM ATP was studied in 0 Na+ Ringer solution; this treatment caused a significant inhibition of dpHi/dt, reversed by 50 æM Bapta. We have shown that H+-ATPase present in MDCK C11 cells depends on Cl- ions and their channels, being regulated by cAMP and ATP, but not by the electrical gradient established by electrogenic H+ transport.

Glutathione and the redox control system trypanothione/trypanothione reductase are involved in the protection of Leishmania spp. against nitrosothiol-induced cytotoxicity

Glutathione is the major intracellular antioxidant thiol protecting mammalian cells against oxidative stress induced by oxygen- and nitrogen-derived reactive species. In trypanosomes and leishmanias, trypanothione plays a central role in parasite protection against mammalian host defence systems by recycling trypanothione disulphide by the enzyme trypanothione reductase. Although Kinetoplastida parasites lack glutathione reductase, they maintain significant levels of glutathione. The aim of this study was to use Leishmania donovani trypanothione reductase gene mutant clones and different Leishmania species to examine the role of these two individual thiol systems in the protection mechanism against S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a nitrogen-derived reactive species donor. We found that the resistance to SNAP of different species of Leishmania was inversely correlated with their glutathione concentration but not with their total low-molecular weight thiol content (about 0.18 nmol/10(7) parasites, regardless Leishmania species). The glutathione concentration in L. amazonensis, L. donovani, L. major, and L. braziliensis were 0.12, 0.10, 0.08, and 0.04 nmol/10(7) parasites, respectively. L. amazonensis, that have a higher level of glutathione, were less susceptible to SNAP (30 and 100 µM). The IC50 values of SNAP determined to L. amazonensis, L. donovani, L. major, and L. braziliensis were 207.8, 188.5, 160.9, and 83 µM, respectively. We also observed that L. donovani mutants carrying only one trypanothione reductase allele had a decreased capacity to survive (40 percent) in the presence of SNAP (30-150 µM). In conclusion, the present data suggest that both antioxidant systems, glutathione and trypanothione/trypanothione reductase, participate in protection of Leishmania against the toxic effect of nitrogen-derived reactive species.

Retinopatía diabética 5 años después / Diabetic retinophaty five years later

Determinar la evolución de la Retinopatía Diabética y sus efectos sobre la función visual en un período de 5 años. 76 pacientes diabéticos seguidos por un período de 5 años. Al grupo de 76 pacientes se les práctico una evaluación integral incluyendo agudeza visual, estado del fondo, angiografía fluoresceínica y niveles de Glicemia. El número de pacientes sin retinopatía diabética bajo de 29 por ciento en 1999 a 18.5 por ciento en el año 2005 con incremento de la RDPP de 20 por ciento a 31 por ciento y la RDP de 25 por ciento a 32 por ciento. Se aprecio un importante aumento de los ojos con visión peor a 20/400 de 35 por ciento a 46 por ciento. En el período de evaluación de 5 años, se nota un importante empeoramiento de la retinopatía diabética en la mayoría de los pacientes, así como también un importante deterioro de la visión.

Abordaje endoscópico de las fístulas de líquido cefalorraquídeo / Endoscopic approach of the cerebrospinal fluid fistulas

La fístula de líquido cefalorraquídeo (FLCR) corresponde a una solución de continuidad de la duramadre, de etiología diversa, por la cual drena LCR hacia las cavidades extracraneales. El tratamiento puede ser tanto médico como quirúrgico, siendo este último el que ofrece mejores garantías de éxito. La vía de abordaje endoscópica transnasal posee una serie de ventajas respecto a la técnicas tradicionales, pero tiene sus indicaciones y contraindicaciones. Se revisaron restrospectivamente 12 pacientes sometidos a esta técnica, analizando los resultados obtenidos con un seguimiento hasta 15 meses. Se concluye que la técnica es afectiva y ventajosa para el paciente, lo que hace recomendable su aprendizaje por quienes realizan cirugía endoscópica transnasal.

Fluoresceína na avaliaçäo da viabilidade tecidual em fraturas expostas graves / Fluorescein to evaluate tissue viability in severe open fractures

Estudo clínico experimental usa a fluoresceína para avaliar a viabilidade tecidual em fraturas expostas graves, como marcador para delimitar as áreas que iräo evoluir com necrose. Foram avaliados dez pacientes com fraturas expostas de grau III. Dos quatro pacientes que apresentaram teste positivo (áreas fluorescentes ou intermediárias), nenhum evoluiu com necrose e um teve infecçao; três pacientes apresentaram teste negativo em grandes áreas; todos evoluíram com necrose e infecçäo; e três pacientes que apresentaram teste negativo, em pequenas áreas, evoluíram sem necrose ou infecçäo. Este é um estudo preliminar, havendo necessidade de séries maiores e continuidade da pesquisa. Os autores acreditam que a fluoresceína pode ser usada como adjuvante quando há dúvida na decisäo do desbridamento pelo cirurgiäo.

Estudo comparativo da tonometria de aplanaçäo com e sem anestesia / Comparative study of applanation tonometry with and without anesthesis

A finalidade deste trabalho é verificar o índice de aceitaçäo e confiabilidade das tonometrias de aplanaçäo, realizadas sem anestesia, com ou sem o uso de fluoresceína. Neste estudo foi utilizado uma amostra de 206 pacientes, avaliando-se o grau de sensibilidade ao exame, nível de escolaridade, comprometimento corneano e análise estatística dos resultados, relacionado com a faixa etária dos pacientes. Foi possível realizar as tonometrias de aplanaçäo em 163 pacientes (79,13 por cento), näo sendo possível em 43 (20,87 por cento). A diferença entre as tonometrias de aplanaçäo com ou sem anestesia näo foram significativas (p<0,05). Conclusäo: as tonometrias de aplanaçäo podem ser realizadas sem anestesia tópica e sem o uso de fluoresceína. Há confiabilidade nos resultados com esta técnica, abrevia-se o tempo de exame e evita-se consequências de uso de medicaçöes que poderäo produzir alergia ou serem veículos de germes. A maioria dos pacientes testados permitiu a realizaçäo da tonometria de aplanaçäo com a técnica proposta

Correlación clínico patológica de células caliciformes y el grado de retinopatía diabética / Relationship clinico-pathological of diabetic retinopathy and globet cells

Es un estudio prospectivo donde se estudiaron a 38 pacientes con 73 ojos, de los cuales 41 ojos fueron derechos y 32 izquierdos; los cuales fueron divididos en tres grupos: con retinopatía diabética proliferativa, no proliferativa y un grupo control de la misma edad y sexo. Esto fue para determinar el número de células calciformes (globet), en diabéticos y compararlo con el grupo control para normar cirterio. Se les realizaron los siguientes estudios: tiempo de ruptura lágrimal, prueba de Schirmer I, tinción de rosa de bengala y fluoresceína así como biopsia de conjuntiva para estudio histológico. Encontramos menor número de células caliciformes (globet) en los pacientes diabéticos que en los pacientes sanos. También se encontró un tiempo de ruptura lágrimal acortado en pacientes femeninos diabéticos así como disminución en la calidad de la lágrima y no en la cantidad. Obtuvimos diferencia estadística significativa para la población estudiada y la expresamos como: p<0.05, con una confiabilidad del 95 por ciento.