Calcium homeostasis behavior and cardiac function on left ventricular remodeling by pressure overload

Sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) and sarcolemmal Na+/Ca2+ exchanger (NCX1) structures are involved in heart cell Ca2+ homeostasis. Previous studies have shown discrepancies in their function and expression in heart failure. The goal of this study was to evaluate heart function and hypertrophied muscle Ca2+-handling protein behavior under pressure overload. Twenty male Wistar rats were divided into two groups: Aortic stenosis (AoS), induced by a clip placed at the beginning of the aorta, and Control (Sham). After 18 weeks, heart function and structure were evaluated by echocardiogram. Myocardial function was analyzed by isolated papillary muscle (IPM) at basal condition and Ca2+ protein functions were evaluated after post-pause contraction and blockage with cyclopiazonic acid in IPM. Ca2+-handling protein expression was studied by western blot (WB). Echocardiogram showed that AoS caused concentric hypertrophy with enhanced ejection fraction and diastolic dysfunction inferred by dilated left atrium and increased relative wall thickness. IPM study showed developed tension was the same in both groups. AoS showed increased stiffness revealed by enhanced resting tension, and changes in Ca2+ homeostasis shown by calcium elevation and SERCA2a blockage maneuvers. WB revealed decreased NCX1, SERCA2a, and phosphorylated phospholambam (PLB) on serine-16 in AoS. AoS had left ventricular hypertrophy and diastolic dysfunction compared to Sham; this could be related to our findings regarding calcium homeostasis behavior: deficit in NCX1, SERCA2a, and phosphorylated PLB on serine-16.

Effect of doses and administration routes of 9R vaccine on protection of Japanese quails against experimental infection with Salmonella Gallinarum / Efeito das doses e vias de administração da vacina 9R na proteção de codornas japonesas contra infeccção experimental com Salmonella Gallinarum

Coturniculture has increased significantly in the last decades. There are several pathogens that can affect these birds. Among the diseases, fowl typhoid stands out as a disease with a potentially great impact to the poultry industry. The objective of this the study was to evaluate the effect of doses and administration routes of live 9R vaccine on protection of Japanese quails against experimental infection with Salmonella Gallinarum (SG). Two hundred and fifty birds were used, divided into five groups: G1, oral vaccination with one dose; G2, oral vaccination with 2 doses; G3, subcutaneous vaccination with one dose; G4, subcutaneous vaccination with two doses and G5 not vaccinated. All birds from all five groups were challenged with SG at an age of 45 days. SG was quantified in the periods of one, four, seven and twelve days after the challenge. The presence of clinical signs and macroscopic lesions of the disease were observed. The groups vaccinated by subcutaneous route had a higher egg production and lower mortality rate. Birds receiving a dose of the vaccine by subcutaneous route also showed lower amount of SG in the liver and spleen seven days after the challenge.(AU)

A coturnicultura tem aumentado significativamente nas últimas décadas. Existem vários patógenos que podem afetar essas aves. Entre as doenças, o tifo aviário se destaca como uma doença de grande impacto para a indústria avícola. O objetivo deste estudo foi avaliar o efeito de doses e vias de administração da vacina viva 9R na proteção de codornas japonesas contra infecção experimental por Salmonella Gallinarum (SG). Foram utilizadas duzentos e cinquenta aves, divididas em cinco grupos: G1, vacinação oral com uma dose; G2, vacinação oral com 2 doses; G3, vacinação subcutânea com uma dose; G4, vacinação subcutânea com duas doses e G5 não vacinado. Todas as aves dos cinco grupos foram desafiadas com SG aos 45 dias de idade. A SG foi quantificada nos períodos de um, quatro, sete e doze dias após o desafio. Foi observada a presença de sinais clínicos e lesões macroscópicas da doença. Os grupos vacinados por via subcutânea apresentaram maior produção de ovos e menor taxa de mortalidade. Aves recebendo uma dose da vacina por via subcutânea também apresentaram menor quantidade de SG no fígado e baço sete dias após o desafio.(AU)

Cardiac function and intracellular Ca2+ handling proteins are not impaired by high-saturated-fat diet-induced obesity

Obesity is often associated with changes in cardiac function; however, the mechanisms responsible for functional abnormalities have not yet been fully clarified. Considering the lack of information regarding high-saturated-fat diet-induced obesity, heart function, and the proteins involved in myocardial calcium (Ca2+) handling, the aim of this study was to test the hypothesis that this dietary model of obesity leads to cardiac dysfunction resulting from alterations in the regulatory proteins of intracellular Ca2+ homeostasis. Male Wistar rats were distributed into two groups: control (C, n=18; standard diet) and obese (Ob, n=19; high-saturated-fat diet), which were fed for 33 weeks. Cardiac structure and function were evaluated using echocardiographic and isolated papillary muscle analyses. Myocardial protein expressions of sarcoplasmic reticulum Ca2+-ATPase, phospholamban (PLB), PLB serine-16 phosphorylation, PLB threonine-17 phosphorylation, ryanodine receptor, calsequestrin, Na+/Ca2+ exchanger, and L-type Ca2+ channel were assessed by western blot. Obese rats presented 104% increase in the adiposity index (C: 4.5±1.4 vs Ob: 9.2±1.5%) and obesity-related comorbidities compared to control rats. The left atrium diameter (C: 5.0±0.4 vs Ob: 5.5±0.5 mm) and posterior wall shortening velocity (C: 36.7±3.4 vs Ob: 41.8±3.8 mm/s) were higher in the obese group than in the control. The papillary muscle function was similar between the groups at baseline and after inotropic and lusitropic maneuvers. Obesity did not lead to changes in myocardial Ca2+ handling proteins expression. In conclusion, the hypothesis was not confirmed, since the high-saturated-fat diet-induced obese rats did not present cardiac dysfunction or impaired intracellular Ca2+ handling proteins.

Sonographic abnormalities in augmented bladder using porcine intestinal submucosa (SIS) / Anormalidades ultrassonográficas em bexigas submetidas à cistoplastia utilizando submucosa intestinal suína (SIS)

Among the different materials for bladder augmentation, porcine intestinal submucosa (SIS) is the most widely investigated and stands out for its ability as a cell scaffold. In this context, the ultrasound examination allows the detection of changes from the surgical procedure, enabling the early verification of potential complications and evaluation of patient outcomes. The aim of this paper is to describe the main sonographic findings in dogs submitted to cystoplasty using acellular SIS and seeded with homologous smooth muscle cells at 30 (M30) and 60 (M60) days postoperatively. Sonographic changes included irregularities and thickening of bladder wall especially at M30. Additionally, were visualized urinary sediment and uroliths in animals submitted to acellular SIS cistoplasty. Abdominal ultrasonography was useful in the postoperative evaluation of animals undergoing cystoplasty with acellular or seeded SIS.(AU)

Entre os diferentes materiais utilizados para a reparação e o aumento da bexiga, a submucosa intestinal suína (SIS) é o mais amplamente investigado e se destaca por sua habilidade como um arcabouço celular. Nesse contexto, o exame ultrassonográfico possibilita a avaliação pós-operatória, bem como a detecção precoce de potenciais complicações. O objetivo deste artigo é descrever os principais achados ultrassonográficos em cães submetidos à cistoplastia utilizando a SIS acelular e semeada com células musculares lisas homólogas aos 30 (M30) e 60 (M60) dias de pós-operatório. As alterações ultrassonográficas pós-operatórias incluíram irregularidades e espessamento da parede da bexiga, especialmente aos M30. Além disso, foram visualizados sedimento urinário e urólitos em animais submetidos à cistoplastia utilizando a SIS acelular. A ecografia abdominal foi útil na avaliação pós-operatória dos animais submetidos à cistoplastia utilizando a SIS acelular e semeada com células musculares lisas homólogas.(AU)

Mensurações do bulbo ocular e cálculo do poder dióptrico da lente intraocular em miniporcos / Ocular bulb measurements and diopter power calculation of the lens in minipigs

Objetivou-se com este trabalho determinar o poder dióptrico da lente intraocular (LIO) em miniporco e as dimensões do bulbo do olho. Foram utilizados 17 miniporcos, sadios, adultos, machos e fêmeas, com peso médio de 70kg. Em todos os olhos foram realizadas a ultrassonografia modo A, a ceratometria e a medida da distância limbo a limbo. O cálculo do poder dióptrico da LIO foi obtido utilizando-se as fórmulas Haigis, Hoffer Q, SRK/T, Holladay I e Holladay II e o software Holladay IOL Consultant(r). Na comparação entre o sexo e a lateralidade do olho, não houve diferença nas variáveis biométricas e poder da LIO. A aplicação das fórmulas (Haigis, Holladay II, Holladay I, SRK/T e Hoffer Q) possibilitou o cálculo do poder da LIO. A Holladay II, fórmula que melhor individualiza o bulbo do olho do miniporco, estima valor dióptrico ao redor de 41 D. Os miniporcos têm potencial como modelo experimental em oftalmologia, relacionado ao seu menor porte e à facilidade no manejo, especialmente em experimentos de longa duração.

The aim of this study was to determine the refractive power of intraocular lens (IOL) of mini pigs and the dimensions of the eyeball. A total of 17 (34 eyes) healthy, adult, males and female animals, with average weight of 70kg were used. For every eye, A-mode ultrasound, keratometry and the measurement of limbo-to-limbo distance were conducted, all variables for calculating the refractive power of the IOL. The value was obtained using different formulas and Holladay IOL Consultant(r) Software. Additionally, the ocular measurements were compared per sex, laterality of the eye and the different formulas used in this study (Haigis, Hoffer Q, SRK / T, Holladay I and Holladay II). In the comparison between sex and laterality of the eye, there was no difference in biometric variables and power of the IOL. The application of the employed formulas (Haigis, Holladay II, Holladay I, SRK / T and Hoffer Q) allowed the IOL power calculation for this specie, and the observed value ranged between 39.58±2.15 and 46.60±2.81 diopters. Mini pigs play an important and growing role as an experimental model for study and practice of ophthalmic procedures, specially related to their smaller size and easy management in long-term experiments.

Classification of different degrees of adiposity in sedentary rats

In experimental studies, several parameters, such as body weight, body mass index, adiposity index, and dual-energy X-ray absorptiometry, have commonly been used to demonstrate increased adiposity and investigate the mechanisms underlying obesity and sedentary lifestyles. However, these investigations have not classified the degree of adiposity nor defined adiposity categories for rats, such as normal, overweight, and obese. The aim of the study was to characterize the degree of adiposity in rats fed a high-fat diet using cluster analysis and to create adiposity intervals in an experimental model of obesity. Thirty-day-old male Wistar rats were fed a normal (n=41) or a high-fat (n=43) diet for 15 weeks. Obesity was defined based on the adiposity index; and the degree of adiposity was evaluated using cluster analysis. Cluster analysis allowed the rats to be classified into two groups (overweight and obese). The obese group displayed significantly higher total body fat and a higher adiposity index compared with those of the overweight group. No differences in systolic blood pressure or nonesterified fatty acid, glucose, total cholesterol, or triglyceride levels were observed between the obese and overweight groups. The adiposity index of the obese group was positively correlated with final body weight, total body fat, and leptin levels. Despite the classification of sedentary rats into overweight and obese groups, it was not possible to identify differences in the comorbidities between the two groups.

Avaliação histomorfométrica e da proliferação celular em úlceras de córnea superficiais, induzidas em coelhos, após o uso de colírio de Citrus lemon / Histomorphometry evaluation and cellular proliferation in superficial corneal ulcer, induced in rabbits, after using eyedrops of Citrus lemon

O objetivo do presente estudo foi avaliar e comparar, por meio de histomorfometria e imuno-histoquímica para PCNA (Antígeno Nuclear de Proliferação Celular), o processo de reparação corneal de úlceras superficiais induzidas em coelhos e tratadas com colírios de óleo essencial de Citrus lemon (CL). Foram utilizadas 40 fêmeas da espécie leporina, constituindo-se quatro grupos experimentais de 10 animais cada. Todos os animais foram submetidos à indução da úlcera superficial experimental por meio da aplicação tópica de n-heptanol. Em dois grupos foram instilados colírios à base de óleo essencial de Citrus lemon, em diferentes concentrações, sendo 3% (GL3) e 5% (GL5). Outro grupo foi tratado com Tween 80 8% (GT), que é o diluente utilizado na produção dos colírios de CL; o grupo controle (GC) recebeu apenas substituto da lágrima. Todos os colírios foram aplicados quatro vezes ao dia. Os grupos experimentais foram distribuídos em dois subgrupos, com cinco animais cada, de acordo com os períodos finais de avaliação. O primeiro subgrupo (M1) foi avaliado após 24 horas e o segundo (M2), após cinco dias. Nas comparações entre os momentos iniciais e finais, os grupos tratados com substituto da lágrima, Tween 80 8% e colírio à base de óleo essencial de Citrus lemon 5% promoveram aumento na espessura epitelial na periferia da córnea e maior percentual de proliferação celular. Não houve diferença de celularidade entre os tratamentos. Os colírios à base de óleo essencial de Citrus lemon, nas diferentes concentrações, promoveram a reepitelização corneal, sem causar lesões adicionais ao epitélio ou estroma corneal, podendo ser utilizado na superfície ocular.

The aim of this study was to evaluate and compare through histomorphometry and immunohistochemistry for PCNA (Proliferating Cell Nuclear Antigen), the repair process in superficial corneal ulcers induced in rabbits and treated with eyedrops of Citrus lemon (CL) essential oil. Fifty female rabbits were used and divided into 4 experimental groups of 10 animals each one. Every animal underwent induction of experimental superficial ulcer by topical application of n-heptanol. Three groups were treated with eyedrops of Citrus lemon essential oil in two different concentrations: 3% (GL3) and 5% (GL5). Another group was treated with Tween 80 8% (GT), which is the solvent used in the production of eyedrops of CL; the control group (CG) received only tear substitute. All eyedrops were applied four times daily. The experimental groups were divided into two subgroups with five animals in each one, according to the final evaluation periods. The first subgroup (M1) was evaluated after 24 hours and the second (M2) after 5 days. In the comparison between the initial and final moments, the groups treated with tear substitute, Tween 80 8% and eyedrops of Citrus lemon essential oil 5% had an increase in epithelial thickness at the periphery of the cornea and a higher percentage of cell proliferation. There was no difference in cellularity between treatments. The eyedrops of Citrus lemon essential oil, at different concentrations, promoted corneal reepithelialization without causing further injury to the epithelium and corneal stroma, so they can be used on the ocular surface.

Expressão de metaloproteinases de matriz e PCNA em úlceras de córnea profundas, induzidas em coelhos, tratadas com plasma rico em plaquetas / Expression of matrix metalloproteinases and PCNA in deep corneal ulcers induced in rabbits, treated with platelet-rich plasma

O objetivo deste estudo foi avaliar a influência do plasma rico (PRP) e pobre (PPP) em plaquetas na proliferação celular e expressão de metaloproteinases de matriz (MMPs), durante a reparação de úlceras corneais profundas. Foram utilizadas 45 coelhas, distribuídas em 3 grupos (G) experimentais (n=15), designados como grupos PRP (GR), PPP (GP) e Controle (GC), de acordo com o tratamento. Todos os animais foram submetidos à indução cirúrgica unilateral de úlcera corneal. No GR e GP, o sangue autólogo foi centrifugado, utilizando-se protocolo padronizado, e foram confeccionados os colírios de PRP e PPP, e instilados cinco vezes ao dia. No GC, foi utilizado colírio lubrificante. Cada grupo foi subdividido (n=5), segundo o momento final de avaliação, sendo 4 (M4), 7 (M7) e 30 dias (M30). As córneas dos animais foram processadas para avaliação morfológica e imuno-histoquímica para PCNA, MMP1, MMP2, MMP9, MT1-MMP e TIMP1. No M4, os níveis de MMP2 foram maiores no GP e GR, sendo que, no M7, esse comportamento foi observado apenas no GP. No M30, no GR, verificou-se maior número de células epiteliais e marcação para MMP1 que o GP. No GR, a proliferação celular foi maior no M4 que nos demais momentos, e a marcação para MMP2 foi maior no M4 que no M30. O PRP estimula a proliferação celular na fase inicial (M4) do tratamento quando comparado aos demais momentos, diferentemente dos demais tratamentos. O uso de colírios de plasma rico e pobre em plaquetas influencia a expressão de metaloproteinases de matriz envolvidas no processo de reparação corneal.

The aim of this study was to evaluate the influence of platelet-rich (PRP) and poor (PPP) plasma in cell proliferation and matrix metalloproteinases (MMPs) expression during the repair of deep corneal ulcers. Forty-five female rabbits were distributed in 3 experimental groups (G) (n = 15), referred to as PRP (GR), PPP (GP) and Control (GC) groups, in accordance with the treatment. All animals underwent surgical induction of unilateral corneal ulcer. PRP and PPP eye drops were made by using centrifuged blood through standardized protocol, and instilled five times a day. In GC, lubricant eye drops were used. Each group was subdivided (n = 5) according to the final time point, 4 (M4), 7 (M7) and 30 days (M30). The animals' corneas were processed for morphological and immunohistochemical analysis for PCNA, MMP1, MMP2, MMP9, MT1-MMP and TIMP1. In M4, the levels of MMP2 were higher in GP and GR, and in M7, this behavior was only observed in the GP. In M30, more epithelial cells and MMP1 expression were found in GR than GP. In GR, cell proliferation was higher in M4 than at other time points and MMP2 expression was higher in M4 than M30. The PRP stimulates cell proliferation in the early phase (M4) of treatment when compared to other time points, different from other treatments. The use of eye drops of platelet-rich and poor plasma influences the expression of matrix metalloproteinases involved in the corneal repair process.

Avaliação histopatológica da cápsula posterior associada ao implante de lente intraocular com superfície modificada com plasma de flúor e polietilenoglicol em coelhos / Histopathological evaluation of posterior capsule associated with intraocular lens implant with modified surface using fluorine and poliethilene glycol plasma in rabbits

O objetivo deste estudo foi avaliar o efeito do tratamento da superfície de lentes intraoculares acrílicas utilizando-se plasma de flúor ou polietilenoglicol na prevenção da opacidade de cápsula posterior. Foram analisados 40 olhos de coelhos, submetidos à cirurgia de facoemulsificação e distribuídos em quatro grupos experimentais (n=10), sendo estes: grupo controle, coelhos sem implante de lente intraocular; grupo com lente intraocular tratada com plasma de polietilenoglicol; grupo com lente intraocular tratada com plasma de flúor; e grupo com lente intraocular comercial. As cápsulas posteriores das lentes dos grupos foram avaliadas por meio de análise histopatológica (morfometria e imuno-histoquímica). Os grupos com lente intraocular tratada com polietilenoglicol e com lente intraocular comercial apresentaram menor espessura da cápsula posterior na avaliação inicial (12 semanas) em relação ao grupo controle. No período final de avaliação (6 meses), os tratamentos da superfície da lente intraocular à base de plasma de flúor e polietilenoglicol não reduziram o desenvolvimento das alterações histológicas associadas à opacidade de cápsula posterior. O tratamento das superfícies das lentes intraoculares com plasma de flúor e polietilenoglicol pode ser realizado como adjuvante na prevenção da opacidade de cápsula posterior, pois não causa alterações na morfologia da lente após facoemulsificação.

The aim of this study was to evaluate the effect of surface treatment of acrylic intraocular lens using Fluorine plasma or polyethylene glycol in the prevention of posterior capsule opacification. Forty rabbit eyes that underwent phacoemulsification were analyzed and distributed into four experimental groups (n=10): Control group, composed of rabbits without intraocular lens implantation; intraocular lens group treated with Polyethylene glycol plasma, intraocular lens group treated with Fluoride plasma, and commercial intraocular lens group. The posterior capsule of the lens was evaluated by histopathological analysis, including morphometric and immunohistochemical studies. Groups with intraocular lens treated with polyethylene glycol and commercial intraocular lens presented thinner posterior capsule at initial assessment (12 weeks) compared to the control group. At the end of the evaluation (six months), the treatment of surface intraocular lens with fluorine and polyethylene glycol plasma did not reduce the development of histological changes associated with posterior capsule opacification. The surface treatment of the intraocular lens with Fluoride and Polyethylene glycol can be performed as an adjuvant in preventing posterior capsule opacification, because it does not cause changes in the morphology of lens after phacoemulsification surgery.

Uso de plasma rico em plaquetas em úlceras de córnea em cães / Use of platelet rich plasma on corneal ulcers in dogs

Este trabalho teve por objetivo avaliar, clínica e macroscopicamente, o tratamento adjuvante com plasma rico em plaquetas na forma de colírio ou tampão, em úlceras de córnea de cães atendidos no Serviço de Oftalmologia Veterinária. Foram analisados 20 olhos com diagnóstico de ceratite ulcerativa, distribuídos em dois grupos experimentais. O grupo colírio (GC) foi constituído por olhos tratados topicamente com colírio autólogo de plasma rico em plaquetas (PRP), e o grupo tampão (GT) por olhos submetidos ao tratamento à base de tampão sólido de PRP, associado ao recobrimento com terceira pálpebra para retenção deste. Os grupos foram avaliados, por meio de avaliação clínica, macroscópica e análise da redução do defeito epitelial, em diferentes momentos, aos três, cinco, dez, 15 e 30 dias, com exceção do terceiro dia no GT. O recobrimento da terceira pálpebra foi removido no quinto dia no GT. Em ambos os grupos, houve redução dos sinais de inflamação, melhora na sensibilidade ocular e adequada reparação do defeito epitelial. Todos os olhos do GT apresentaram completa cicatrização no quinto dia e 70% no GC, atingindo a totalidade no 10º dia. O PRP na forma de colírio ou tampão é uma excelente terapia adjuvante a ser instituída no tratamento clínico da úlcera de córnea em cães, pois atua na diminuição dos sinais inflamatórios, da dor ocular e auxilia potencialmente na cicatrização do defeito epitelial...

This study aims to clinically and macroscopically evaluate the adjuvant therapy with platelet-rich plasma in the form of eyedrops or clot, for corneal ulcers in dogs treated at the Veterinary Ophthalmology Service. We analyzed 20 eyes diagnosed with ulcerative keratitis, divided into two experimental groups. The eyedrop group (GC) was composed of eyes treated topically with eyedrops of autologous platelet-rich plasma (PRP), and the clot group (GT) was composed of eyes treated with a platelet-rich clot and covered with a third eyelid for retention of the clot. The groups were evaluated by clinical and macroscopic analysis and by the analysis of epithelial defect reduction, at different times, at three, five, ten, 15 and 30 days, except for the third day in GT. The coverage of the third eyelid was removed on the fifth day. In both groups the inflammation signs reduced, there was an improvement in ocular sensibility and proper repair of epithelial defect. All GT eyes and 70% GC eyes showed complete healing on the fifth day, the remainder of GC completed healing on the tenth day. PRP in the form of eyedrops and clot is an excellent adjuvant therapy to be instituted in the clinical treatment for corneal ulcer in dogs, because it decreases the inflammatory signs and the ocular pain and it potentially assists in healing epithelial defects...

Uso do plasma rico em plaquetas no reparo de úlceras de córnea profundas induzidas em coelhos. Avaliação clínica e histomorfométrica / Use of platelet-rich plasma in the treatment of deep corneal ulcers induced in rabbits. Clinical and histomorphometric evaluation

Avaliou-se e comparou-se, clinicamente, por meio de histopatologia e morfometria, o processo de reparação corneana de úlceras experimentais induzidas em coelhos, com o uso de plasma rico em plaquetas (PRP) sob a forma de colírio ou tampão. Foram utilizados 60 coelhos, constituindo-se quatro grupos experimentais de 15 animais cada, designados grupo plaqueta (GP), grupo tampão (GT), grupo controle (GC) e grupo controle amniótica (GA). Os grupos experimentais foram subdivididos em três subgrupos (M4, M7, M30), de acordo com o período final de avaliação, aos quatro, sete e 30 dias, respectivamente. Não houve diferenças entre os tratamentos utilizados quanto aos sinais relacionados à sensibilidade ocular, quemose e secreção ocular. Os grupos tratados com PRP, na forma de tampão ou colírio, apresentaram menor opacidade corneana do que os animais tratados apenas com membrana amniótica no momento final de avaliação. Quanto à presença da úlcera corneana, os grupos tratados com PRP apresentaram menor ulceração corneana em relação aos demais grupos. No exame histomorfométrico, verificou-se maior epitelização corneana na fase inicial da lesão no tratamento à base de colírio de plasma rico em plaquetas. O uso da membrana amniótica promoveu espessamento do epitélio e estroma corneano, com sinergismo dela quando associada ao plasma rico em plaquetas.

The repair process induced corneal ulcer in rabbits using platelet-rich plasma in the form of eyedrop or clot was clinically evaluated and compared. Sixty rabbits were divided into four groups of 15 animals, denominated platelet group (PG), clot group (CLG), control group (CG), and amniotic control group (AG). Experimental groups were then subdivided into three groups (M4, M7, M30), corresponding to the end of the evaluation period. There were no differences between treatments regarding ocular sensitivity, chemosis and ocular secretion. The groups treated with PRP either as eyedrop or a clot showed less opacity than the animals treated only with amniotic membrane at the moment of the final evaluation. The presence of corneal ulcers in the groups treated with PRP showed lower intensity than the other groups. Histomorphometric examination showed that corneal epithelization in the initial phase of the lesion was greater when using PRP. The use of amniotic membrane promoted corneal epithelial and stromal thickness, as well as synergism when associated to PRP.

Implante de condrócitos homólogos em defeitos osteocondrais de cães: padronização da técnica e avaliação histopatológica / Homologous articular chondrocytes implantation in osteochondral defects of dogs: technique and histopathological evaluation standardization

Padronizou-se a metodologia para cultura de condrócitos em cães e avaliou-se seu implante em lesões osteocondrais, utilizando-se a membrana biossintética de celulose (MBC) como revestimento. Dez cães, adultos e clinicamente sadios, foram submetidos à artrotomia das articulações fêmoro-tíbio-patelares. Defeitos de 4mm de diâmetro e profundidade foram induzidos no sulco troclear de ambos os membros. MBC foi aplicada na base e na superfície das lesões. Os defeitos do membro direito foram preenchidos com condrócitos homólogos cultivados formando o grupo-tratado (GT); os do membro esquerdo, sem implante celular, foram designados grupo-controle (GC). A evolução pós-operatória foi analisada com especial interesse nos processos de reparação da lesão, por meio de histomorfometria e imuno-histoquímica para colágeno tipo II e sulfato de condroitina. A cultura de condrócitos homólogos apresentou alta densidade e taxa de viabilidade. Observou-se integridade do tecido neoformado com a cartilagem adjacente na avaliação histológica, em ambos os grupos. Na imuno-histoquímica, verificou-se predomínio de colágeno tipo II no GT. Morfometricamente, não houve diferença significativa entre o tecido fibroso e o fibrocartilaginoso entre os grupos. A cultura de condrócitos homólogos de cães foi exequível. O tecido neoformado apresentou qualidade discretamente superior associado ao implante homólogo de condrócitos, contudo não promoveu reparação por cartilagem hialina.

The aim of the study is to standardize the methodology to achieve canine chondrocytes culture, and evaluate its implant on osteochondral defects made in the femoral trochlear sulcus of dogs, using the cellulose biosynthetic membrane (CBM) as coating. Ten healthy adult dogs without locomotor disorders were used. All animals were submitted to arthrotomy of stifle joints and defects of four millimeters in diameter x four millimeters deep were done in the femoral trochlear sulcus of both limbs. CBM were applied in the lesion base and surface of all limbs. In the treated group (TG), defects of the right limb were filled with cultivated homologous chondrocytes, and in control group (CG), defects of the left limb were left without cellular implant. Postoperative follow up was done by histomorphometry and Collagen type II and anti-chondroitin sulfate immunohistochemistry. The homologous chondrocytes culture showed high density and viability rate. Upon immunohistochemistry the predominance of type II collagen in extracellular matrix of TG was verified. However, no significant statistical difference was observed between the groups upon histomorphometry analysis of fibrous and fibrocartilaginous tissues. Canine homologous chondrocytes culture was practicable. Neoformed tissue showed slightly higher quality in TG, but without promoting repair by the hyaline cartilage.

Confiabilidade e pontuação mínima relacionada à intervenção analgésica de uma escala multidimensional para avaliação de dor pós-operatória em gatos / Reliability and cut-off point related to the analgesic intervention of a multidimensional composite scale to assess postoperative pain in cats

Avaliou-se a confiabilidade e determinou-se a pontuação mínima relacionada à necessidade de intervenção analgésica de uma escala multidimensional para avaliação de dor pós-operatória em gatas submetidas à ovário-histerectomia. Trinta gatas foram filmadas em quatro momentos predeterminados: pré-operatório, pós-operatório antes e depois da analgesia de resgate e às 24 horas após a cirurgia. Quatro observadores, não cientes dos momentos de avaliação, e um observador, considerado padrão-ouro - pesquisador que desenvolveu a escala - , avaliaram os vídeos em ordem aleatória, a partir da aplicação da escala, e determinaram a suficiência ou não de analgesia, baseando-se na experiência clínica. Os vídeos foram reavaliados dois meses após a primeira análise. O grau de concordância entre as avaliações dos diferentes observadores e das avaliações feitas pelo mesmo observador em diferentes ocasiões foi verificado pelo coeficiente de correlação intraclasse, que variou entre bom e muito bom para todos os itens da escala, exceto atividade, atitude e miscelânea de comportamentos que apresentaram confiabilidade moderada. O ponto de corte ótimo (> 7) foi selecionado pela análise descritiva e da curva Receiver Operating Characteristic. Os resultados suportam a confiabilidade em termos de reprodutibilidade e estabilidade e indicam que a terapia analgésica deve ser considerada em pontuações >8.

The aim with this study was to evaluate the reliability, and to establish the minimum score relating to the analgesic intervention of a multidimensional scale for use in assessing postoperative pain in cats undergoing ovariohysterectomy. Thirty cats were video recorded at 4 predetermined moments: prior to surgery, before and after rescue analgesia postoperatively, and 24 hours after surgery. Four observers blind to the times of assessment, and a gold standard observer (researcher who developed the scale) evaluated the video segments in random order using the scale, and determined the adequacy or not of analgesia based on clinical experience. The videos were reassessed two months after the first analysis. The agreement between the assessment of different observers, and the assessment made by the same observer on different occasions has been verified by the intra-class correlation coefficient that ranged from good to very good for all scale items, except activity, attitude and miscellaneous behaviors, which showed moderate reliability. The optimal cutoff point (> 7) was selected based on descriptive analysis and Receiver Operating Characteristic curve. The findings support the reliability in terms of reproducibility and stability, and indicate that analgesic therapy should be considered in scores >8.

Reparação de defeitos osteocondrais de cães com implante de cultura de condrócitos homólogos e membrana biossintética de celulose: avaliação clínica, ultrassonográfica e macroscópica / The repair of osteochondral defects in dogs with homologous articular chondrocytes and biosynthetic cellulose membrane: clinical, ultrasound and macroscopic evaluation

Avaliou-se o implante de condrócitos homólogos em lesões osteocondrais, utilizando a membrana biossintética à base de celulose (MBC) como revestimento. Dez cães adultos e clinicamente sadios foram submetidos à artrotomia das articulações fêmoro-tíbio-patelares. Defeitos de quatro milímetros de diâmetro por quatro milímetros de profundidade foram induzidos na tróclea femoral de ambos os membros. A MBC foi aplicada na base e superfície das lesões. Os defeitos do membro direito foram preenchidos com condrócitos homólogos cultivados e formaram o grupo tratado (GT); e os defeitos do membro esquerdo, sem implante celular, formaram o grupo controle (GC). Os animais foram avaliados clínica e ultrassonograficamente aos 30 e 60 dias. A evolução pós-operatória dos cães foi analisada com especial interesse nos processos de reparação da lesão, por meio de macroscopia. Não houve diferença clínica e ultrassonográfica entre os grupos. Entretanto, à macroscopia, ocorreu maior prevalência de formação de tecido cicatricial esbranquiçado no GT. O tecido neoformado apresentou melhor qualidade associado ao implante homólogo de condrócitos, mas não promoveu reparação por cartilagem hialina.

The aim of the study was to evaluate the repair of deep cartilaginous defects made in the femoral trochlear sulcus of dogs, using the cellulose biosynthetic membrane (CBM) as coating. Ten healthy adult dogs without locomotor disorders were used. All animals were submitted to arthrotomy of stifle joints and defects with four millimeters diameter x four millimeters deep were done in the femoral trochlear sulcus of both limbs. CBM was applied in the lesion's base and surface of all limbs. In the treated group (TG), the defects of the right limb were filled with cultivated homologous chondrocytes, and in control group (CG), the defects of the left limb were filled without cellular implant. The animals were evaluated by physical examination and ultrasound at 30 and 60 days. The postoperative follow up of the dogs was done by macroscopy with special interest in the healing process of the osteochondral defect. No clinical and ultrasonographic differences were observed in both groups. In the macroscopic evaluation higher prevalence of whitish scar tissue formation was noted in TG, but without statistical difference. The neoformed tissue showed slightly higher quality in TG, but without promoting repair by the hyaline cartilage.

Validade e responsividade de uma escala multidimensional para avaliação de dor pós-operatória em gatos / Validity and responsiveness of a multidimensional composite scale to assess postoperative pain in cats

O manejo adequado da dor depende de métodos de avaliação acurados. O objetivo deste estudo foi avaliar a validade e a responsividade de uma escala multidimensional para avaliação de dor pós-operatória em gatas submetidas à ovário-histerectomia. Trinta gatas foram filmadas no período perioperatório em quatro momentos predeterminados: pré-operatório, pós-operatório antes e depois da analgesia de resgate e às 24 horas após o término da cirurgia. Cinco observadores não cientes dos momentos de avaliação e um observador considerado padrão-ouro (pesquisador que desenvolveu a escala) avaliaram os vídeos em ordem aleatória e determinaram os escores de dor dos animais, utilizando a escala a ser validada. A responsividade e a validade de conteúdo e constructo foram estabelecidas pelo aumento e pela redução significativa nos escores de dor, em resposta ao procedimento cirúrgico e à administração de analgesia de resgate, respectivamente, avaliados pelo teste de Wilcoxon (P<0,001). A validade de critério foi atestada pela concordância alta entre as avaliações do padrão-ouro com os demais observadores, analisada pela porcentagem de absoluta concordância e coeficiente Kappa ponderado. Os resultados suportam a validade e a responsividade de uma escala multidimensional para avaliação de dor aguda pós-operatória em gatas submetidas à ovário-histerectomia, quando utilizada por veterinários formados e com experiência clínica.

Adequate pain management depends on accurate assessment methods. The aim of this study was to evaluate the validity and responsiveness of a multidimensional composite scale for use in assessing postoperative pain in cats undergoing ovariohysterectomy. Thirty cats were video recorded in four predetermined moments during the perioperative period: prior to surgery, before and after rescue analgesia postoperatively, and 24 hours after the end of surgery. Five observers blind to the times of assessment, and a gold standard observer (researcher who developed the scale) viewed the video segments in a random order, and the cat's pain scores were registered using the scale to be validated. The content and construct validity, and the responsiveness were established based on significant increase and reduction in the pain scores, in response to surgery and analgesics administration, respectively (Wilcoxon signed rank test, P<0.001). Criterion validity was determined by high agreement between the evaluations of the gold standard with the other observers, assessed by the percentage of absolute agreement and weighted Kappa coefficient. The findings support the validity and the responsiveness of a multidimensional composite scale to assess postoperative acute pain in cats undergoing ovariohysterectomy, when used by veterinarians with clinical experience.

Uso do colírio de Citrus lemon em úlceras de córnea induzidas em coelhos / Use of Citrus lemon eyedrops in corneal ulcers induced in rabbits

Avaliaram-se e compararam-se clinicamente os efeitos de colírios de Citrus lemon (CL) na reparação de úlcera corneal superficial. Foram utilizadas 50 coelhas, distribuídas em cinco grupos experimentais de 10 animais cada. Após a indução da úlcera, três grupos foram tratados com colírios de CL, nas concentrações de 1,5% (GL1,5), 3% (GL3) e 5% (GL5). Um quarto grupo foi tratado com Tween 80 a 8%, diluente utilizado na produção dos colírios de CL, e o quinto grupo, controle, recebeu substituto da lágrima. Os grupos foram distribuídos em dois subgrupos de cinco animais; assim, o primeiro (M1) foi avaliado após 24 horas, e o segundo (M5) após cinco dias. Não houve diferença entre os tratamentos quanto aos sinais clínicos secreção, hiperemia conjuntival, quemose e opacidade corneal. A extensão da úlcera corneal diminuiu, de forma significativa, em todos os grupos no M5, com exceção do GL3, o qual apresentou menor mediana quanto à extensão no M1. O colírio de óleo essencial de Citrus lemon pode ser utilizado no tratamento de ceratites ulcerativas superficiais, sem induzir irritação conjuntival, favorecendo a reepitelização corneal.

The aim of this study was to clinically evaluate and compare the effects of Citrus lemon (CL) eye drops in the repair of superficial corneal ulcers. Fifty female rabbits were used, constituting five experimental groups with 10 animals each. After ulcer induction, three groups were treated with CL eye drops, at 1.5% (GL1, 5), 3% (GL3) and 5% (GL5) concentrations. Another group was treated with Tween 80 8% solvent used in the production of CL eye drops, and the control group received tear substitute. The experimental groups were divided into two subgroups (five animals). The first (M1) was evaluated after 24 hours and the second (M5) after five days. There was no difference between treatments for clinical signs of secretion, conjunctival hyperemia, chemosis and corneal opacity. The extension of corneal ulcer significantly decreased in all groups in M5, the exception was GL3, which had a lower median of extension in the M1. The eye drops made of Citrus lemon essential oil can be used in the treatment of superficial ulcerative keratitis, without inducing conjunctival irritation, favoring corneal reepithelialization.

Transplante de glândulas salivares menores no tratamento da ceratoconjuntivite seca em cães / Transplantation of the minor salivary gland in dogs with keratoconjunctivitis sicca

Relatou-se o efeito do transplante de glândulas salivares menores (TGSM) em cães portadores de ceratoconjuntivite seca (CCS) e estudaram-se os efeitos da secreção dessas glândulas usadas como alternativa de lubrificação ocular. A aplicação da técnica foi satisfatória, uma vez que resultou em melhora no quadro clínico oftalmológico sem que houvesse mínimas intercorrências pós-operatórias.

The effect of the transplant of minor salivary gland (TGSM) in carrying dogs of dry ceratoconjuntivite (CCS) and the effect of these used glands as alternative for ocular lubrication have been studied. The transplantation of the salivary gland in dogs with keratoconjunctivitis sicca (KCS) was reported and clinical effects of the salivary secretion as ocular lubricant for the relief of the disease were studied. Based on these results, it was concluded that the application of the TMSG technique in dogs with KCS is satisfactory, as demonstrated by improvement in the clinical ophthalmology and appreciated by the landlord regardin the results of the surgery and minimal postoperative complications.

Uso de viscoelásticos na facoemulsificação em cães portadores de catarata: efeitos sobre a pressão intraocular, a morfologia das células endoteliais e a espessura corneana / Use of viscoelastic substances for the phacoemulsification in dogs with cataract: effects on the intraocular pressure, morphology of endothelial cells, and corneal thickness

Avaliaram-se as células endoteliais, a espessura corneana e a pressão intraocular (PIO) de cães portadores de catarata madura, empregando-se viscoelástico à base de hialuronato de sódio 3 por cento e sulfato de condroitina 4 por cento e hidroxipropilmetilcelulose 2 por cento, utilizando-se 20 cães, distribuídos entre os dois grupos dos viscoelásticos. A técnica cirúrgica adotada foi a da facoemulsificação bimanual. As avaliações tonométricas foram efetuadas antes e após o ato cirúrgico, aos 1, 7, 14, 21, 28 e 60 dias de pós-operatório, e a microscopia especular, antes e após 7, 28 e 60 dias. Não houve diferença estatística entre os grupos quanto à PIO, com exceção aos 14 dias, em que se observou maior PIO com o uso de hialuronato de sódio 3 por cento e sulfato de condroitina 4 por cento. Não houve diferença entre os grupos quanto aos parâmetros relacionados ao endotélio, com diminuição discreta da densidade celular endotelial e aumento da área celular com a utilização de hidroxipropilmetilcelulose 2 por cento. A utilização de ambos os dispositivos viscoelásticos analisados é recomendada para o procedimento de facoemulsificação em cães.

The endothelial cells, the corneal thickness, and the intraocular pressure (IOP) were evaluated in dogs with cataract, using viscoelastic substances based on 3 percent sodium hyaluronate and 4 percent chondroitin sulfate and comparing them with 2 percent hydroxypropylmethylcellulose. Twenty dogs were distributed in two groups of ten, each using one viscoelastic material. The surgical technique was bimanual phacoemulsification. The tonometric evaluations were made before and at one, seven, 14, 21, 28, and 60 days after the surgery and the specular microscopy before and after seven, 28, and 60 days. No statistical difference between groups was found according to IOP, except at 14 days, which was significantly higher with the use of 3 percent sodium hyaluronate and 4 percent chondroitin sulfate. There was no statistical difference between the groups considering the parameters related to the endothelium, with slight decrease in endothelial cell density and increase of cell area with the use of 2 percent hydroxypropylmethylcellulose. The use of both viscoelastic devices is recommended for the phacoemulsification in dogs.

Performance of IFAT, ELISA, direct parasitological examination and PCR on lymph node aspirates for canine visceral leishmaniasis diagnosis

Canine visceral leishmaniasis (CVL) is endemic in numerous Brazilian regions. The greatest difficulty in controlling the disease is the diagnostic limitation. In the present study, the most common tests employed for visceral leishmaniasis diagnosis were compared: immunofluorescence antibody test (IFAT), immunoenzymatic assay (ELISA), direct parasitological examination and polymerase chain reaction (PCR). Samples of lymph node aspirates and blood were collected from 100 dogs that lived in an endemic area (Bauru city, São Paulo state) and from 100 negative controls from a non-endemic area (Botucatu city, São Paulo state). Specificity of both IFAT and PCR was 100 percent whereas ELISA was 99 percent. Sensitivities were 97.77, 93.33 and 91.11 percent respectively for IFAT, ELISA and PCR.

Demographic and metabolic characteristics of individuals with progressive glucose tolerance

We evaluated changes in glucose tolerance of 17 progressors and 62 non-progressors for 9 years to improve our understanding of the pathogenesis of type 2 diabetes mellitus. Changes in anthropometric measurements and responses to an oral glucose tolerance test (OGTT) were analyzed. We identified 14 pairs of individuals, one from each group, who were initially normal glucose tolerant and were matched for gender, age, weight, and girth. We compared initial plasma glucose and insulin curves (from OGTT), insulin secretion (first and second phases) and insulin sensitivity indices (from hyperglycemic clamp assay) for both groups. In the normal glucose tolerant phase, progressors presented: 1) a higher OGTT blood glucose response with hyperglycemia in the second hour and a similar insulin response vs non-progressors; 2) a reduced first-phase insulin secretion (2.0 ± 0.3 vs 2.3 ± 0.3 pmol/L; P < 0.02) with a similar insulin sensitivity index and a lower disposition index (3.9 ± 0.2 vs 4.1 ± 0.2 µmol·kg-1·min-1 ; P < 0.05) vs non-progressors. After 9 years, both groups presented similar increases in weight and fasting blood glucose levels and progressors had an increased glycemic response at 120 min (P < 0.05) and reduced early insulin response to OGTT (progressors, 1st: 2.10 ± 0.34 vs 2nd: 1.87 ± 0.25 pmol/mmol; non-progressors, 1st: 2.15 ± 0.28 vs 2nd: 2.03 ± 0.39 pmol/mmol; P < 0.05). Theses data suggest that β-cell dysfunction might be a risk factor for type 2 diabetes mellitus.