Preliminary study of rabbits as an animal model of mammalian eye transplantation and literature review / Estudo preliminar de coelhos como modelo animal para transplante de olho em mamíferos e revisão da literatura

ABSTRACT Purpose: To describe an innovative animal model of eye transplantation used in rabbits. Methods: six Dutch-belted male rabbits were submitted to lateral orbitotomy in the right eye, wide retrobulbar anatomy exposure, dissection of the structures, identification and distal section of the optic nerve followed by anastomosis either by vicryl (group 1) or fibrin glue (group 2). Electroretinography recording was performed before the section of the optic nerve and every 30 seconds after, to monitor the function of retina. Left eye was used as control group. Results: After optic nerve resection and anastomosis, stable ERG amplitude of the right eye was lost after 302 seconds in group 1 and after 296 seconds on group 2. Left eye kept longer stable ERG amplitude curves. Conclusions: The animal model of whole eye transplantation was effective in describing a novel technique to be used in rabbits, with success of the anatomic procedure. Further studies will clarify the best anastomosis methods and maintenance of function of the receptor organ. Translational relevance: this animal model of whole eye transplantation provides a novel perspective for blind patients and the research models, since we describe a novel mammal animal model. This model can be used as basis of a human model of whole eye transplantation in future studies.

RESUMO Objetivo: Descrever uma técnica cirúrgica inovadora para transplante de olho em um modelo animal em coelhos. Métodos: Seis coelhos machos com Dutch Belted foram submetidos à orbitotomia lateral do olho direito, com ampla exposição da anatomia retrobulbar, dissecção do cone muscular, exposição e secção distal do nervo óptico seguida de anastomose por vicryl (Grupo 1) ou cola de fibrina (Grupo 2). O registro da eletrorretinografia foi realizado antes da secção do nervo óptico e a cada 30 segundos após, para monitorar a função da retina. O olho esquerdo foi usado como grupo controle. Resultados: Após a ressecção do nervo óptico, a estabilidade da amplitude da eletrorretinografia foi perdida no olho direito após 302 segundos no Grupo 1 e após 296 segundos no Grupo 2. O olho esquerdo manteve eletrorretinografia estável por períodos mais longos. Conclusão: O modelo animal de transplante total de olho foi eficaz em descrever uma nova técnica cirúrgica para ser utilizada em laboratório com coelhos, com sucesso do procedimento anatômico. Novos estudos esclarecerão os melhores métodos de anastomose e manutenção da função do órgão receptor.

Intravitreal injection of polysorbate 80: a functional and morphological study / Injeção intravítrea de polissorbato 80: estudo funcional e morfológico

ABSTRACT Objective : to determine the functional and morphological effects at rabbits retina of PS80 concentration used in the preparation of intravitreal drugs. Methods: eleven New Zealand rabbits received a intravitreal injection of 0.1ml of PS80. As control, the contralateral eye of each rabbit received the same volume of saline. Electroretinography was performed according to a modified protocol, as well as biomicroscopy and retina mapping before injection and seven and ten days after. Animals were euthanized in the 30th day and the retinas were analyzed by light microscopy. Results: eyes injected with PS80 did not present clinical signs of intraocular inflammation. Electroretinography did not show any alteration of extent and implicit time of a and b waves at scotopic and photopic conditions. There were no morphological alterations of retinas at light microscopy. Conclusion: intravitreal injection of PS80 in the used concentration for intravitreal drug preparations do not cause any functional or morphological alterations of rabbit retinas. These results suggest that PS80 is not toxic to rabbit retinas and may be safely used in the preparation of new lipophilic drugs for intravitreal injection.

RESUMO Objetivo: determinar os efeitos funcionais e morfológicos na retina de coelhos da concentração de PS80 utilizada na preparação de drogas intravítreas. Métodos: onze coelhos New Zealand receberam injeção intravítrea de 0,1ml de PS80. Como controle, o olho contralateral de cada coelho recebeu o mesmo volume de soro fisiológico. Foram realizados eletrorretinogramas de acordo com o protocolo modificado, biomicroscopia e mapeamento de retina antes da injeção, sete e dez dias depois. Os animais foram sacrificados no 30o dia e as retinas analisadas por microscopia de luz. Resultados: os olhos injetados com PS80 não apresentaram sinais clínicos de inflamação intraocular. O eletrorretinograma não apresentou alteração de amplitude e tempo implícito das ondas a e b nas condições escotópica e fotópica. Não houve alteração morfológica da retina na microscopia de luz. Conclusão: a injeção intravítrea de PS80 na concentração utilizada na preparação de drogas intravítreas não causa alterações funcionais e morfológicas na retina de coelhos. Esses resultados sugerem que o PS80 não é tóxico para a retina de coelhos e pode ser usado com segurança na preparação de novas drogas lipofílicas para injeção intravítrea.

Full-field electroretinogram recorded with skin electrodes in normal adults / Eletrorretinograma de campo total registrado com eletrodos de pele em adultos normais

ABSTRACT Purpose: Alternative recording methods have been tested to allow the electroretinogram (ERG) recording in uncooperative patients and/or patients with palpebral alterations, including recordings with skin electrodes. The purpose of this study was to compare ERG recorded with skin electrodes and well-established microfiber electrodes and to determine normative values of ERG parameters for recording with skin electrodes. Methods: Fifty healthy volunteers (17-26 years; mean 20.63 ± 2.01 years) participated in the study. A gold disk skin electrode was placed on the lower orbital rim of a randomly chosen eye. On the contralateral eye, a microfiber electrode was positioned in the lower conjunctival sac. Gold disc electrodes were positioned at the ipsilateral outer canthus of both eyes acting as reference electrodes for the creation of a potential difference. Two ground electrodes were placed on the lobe of each ear. ERGs were recorded according to the International Society of Clinical Electrophysiology Visual (ISCEV) protocol using the VERIS 5.1.9 system for data acquisition and analysis. Results: Both types of electrodes showed similar wave response morphologies. The implicit time of responses between the two electrodes was comparable. On peak-to-peak amplitude, skin electrode recordings showed an amplitude reduction of 61.4% for rod responses, 61.5% for maximal responses, 46.2% for oscillatory potentials, 57.4% for cone responses, and 54.4% for 30Hz-flicker responses, when compared with microfiber electrode recordings. Based on these findings, normative values for peak-to-peak amplitude and implicit time to be used as a reference for ERGs recorded with skin electrodes were determined. Conclusions: ERGs recorded with skin electrodes presented lower peak-to-peak amplitude compared with microfiber electrodes. However, using appropriate normative values, skin electrodes may be useful for specific target populations such as uncooperative infants and/or patients with ocular surface alterations.

RESUMO Objetivo: O eletrorretinograma de campo total (ERG) em pacientes não colaborativos e/ou com alterações palpebrais pode ser registrado com diferentes métodos de captação de respostas, dentre eles os eletrodos de pele. O objetivo deste estudo foi comparar o eletroretinograma obtido com eletrodos de pele e eletrodos de microfibra, determinando valores normativos para os parâmetros registrados com eletrodos de pele. Métodos: Cinquenta voluntários saudáveis (17-26 anos; média 20,63 ± 2,01) participaram deste estudo. Um eletrodo de cúpula de ouro foi aderido à pele na margem orbital inferior de um dos olhos escolhido ao acaso. No olho contralateral, um eletrodo de microfibra foi posicionado no saco conjuntival inferior. Eletrodos de cúpula de ouro foram posicionados na região lateral de cada olho para a formação da diferença de potencial com o eletrodo ativo. Dois eletrodos terra foram posicionados no lobo de cada orelha. O eletrorretinograma foi registrado de acordo com o protocolo da ISCEV (Sociedade Internacional de Eletrofisiologia Visual), com o sistema VERIS 5.1.9 para aquisição e análise dos registros. Resultados: Os dois tipos de eletrodos apresentaram morfologias de onda similares. O tempo implícito das respostas foi comparável entre os dois tipos de eletrodo. Os registros feitos com eletrodo de pele quando comparados aos registros com eletrodos de microfibra mostraram reduções na amplitude das ondas, de 61,4% para resposta de bastonetes, 61,5% para resposta máxima, 46,2% para potenciais oscilatórios, 57,4% para resposta de cones, e 54,4% para flicker 30Hz. Baseado nestes resultados, foram determinados valores normativos para amplitude e tempo de implícito para ERGs obtidos com eletrodos de pele. Conclusões: ERGs registrados com eletrodos de pele apresentam respostas com amplitudes menores quando comparados aos registrados com eletrodos de microfibra. No entanto, usando valores normativos apropriados, os eletrodos de pele podem ser uma alternativa útil para populações especificas como pacientes não colaborativos e/ou com alterações palpebrais.

The Effects of Alcohol on Visual Evoked Potential and Multifocal Electroretinography

The aim of this study was to investigate the acute effects of ethanol administration on pattern-reversal visual evoked potential (VEP) and multifocal electroretinography (mfERG). Fifteen healthy subjects with no ocular or general disease were recruited. VEP (0.25° pattern sizes) and mfERG with 19 elements in two recording segments were performed before ethanol administration to obtain baseline for each participant. A few days later, the participants visited again for VEP and mfERG measurements after ethanol administration. Ethanol (0.75 g/kg) was administered orally over the course of 30 minutes. VEP and blood alcohol concentration were evaluated one hour after ethanol administration, and mfERG was conducted after pupil dilation. The Wilcoxon signed-rank test was used to compare parameter changes after randomized eye selection. The mean blood alcohol concentration was 0.034% ± 0.05% by volume. VEP revealed a P100 latency delay (109.4 ± 5.3; 113.1 ± 8.2; P = 0.008) after alcohol administration. The P1 implicit time of ring 1 on mfERG showed a trend of shortening after alcohol administration (37.9 ± 1.0; 37.2 ± 1.5; P = 0.048). However, the changes did not show statistical significance after Bonferroni correction. In conclusion, orally administrated ethanol (0.75 g/kg) appears to suppress the central nervous system, but it is not clear whether alcohol intake affects the retina.

Normal clinical electroretinography parameters for poodle, Labrador retriever, Thai ridgeback, and Thai Bangkaew

The purpose of the present study was to establish normal electroretinogram (ERG) parameters using 56 normal eyes of four dog breeds common in Thailand: poodle, Labrador retriever, Thai ridgeback, and Thai Bangkaew. Standard ERG findings were bilaterally recorded using a handheld multi-species ERG unit with an ERG-jet lens electrode for 28 dogs under preanesthesia with diazepam, anesthesia with propofol, and anesthesia maintenance with isoflurane. There were significant differences in the mean values of ERG amplitudes and implicit times among the four dog breeds (p < 0.05) except for the b-wave implicit time of the photopic 30 Hz flicker response with 3 cd.s/m2 (p = 0.610). Out of the four breeds, Thai Bangkaew had the longest implicit time (p < 0.001) of scotopic low intensity responses, b-wave of scotopic standard intensity responses (3 cd.s/m2), a-wave of the higher intensity response (10 cd.s/m2), and a-wave of the photopic single flash response (3 cd.s/m2). For the b/a ratio, only the ratio of the Cone response was significantly different among the different breeds. In this summary, normal ERG parameters for four dog breeds were reported. Data from the investigation supported the hypothesis that determination of breed-specific limits of normality for ERG responses is necessary for individual clinics and laboratories.

Vitreous pharmacokinetics and electroretinographic findings after intravitreal injection of acyclovir in rabbits

OBJECTIVES: Acute retinal necrosis is a rapidly progressive and devastating viral retinitis caused by the herpesvirus family. Systemic acyclovir is the treatment of choice; however, the progression of retinal lesions ceases approximately 2 days after treatment initiation. An intravitreal injection of acyclovir may be used an adjuvant therapy during the first 2 days of treatment when systemically administered acyclovir has not reached therapeutic levels in the retina. The aims of this study were to determine the pharmacokinetic profile of acyclovir in the rabbit vitreous after intravitreal injection and the functional effects of acyclovir in the rabbit retina. METHODS: Acyclovir (Acyclovir; Bedford Laboratories, Bedford, OH, USA) 1 mg in 0.1 mL was injected into the right eye vitreous of 32 New Zealand white rabbits, and 0.1 mL sterile saline solution was injected into the left eye as a control. The animals were sacrificed after 2, 9, 14, or 28 days. The eyes were enucleated, and the vitreous was removed. The half-life of acyclovir was determined using high-performance liquid chromatography. Electroretinograms were recorded on days 2, 9, 14, and 28 in the eight animals that were sacrificed 28 days after injection according to a modified protocol of the International Society for Clinical Electrophysiology of Vision. RESULTS: Acyclovir rapidly decayed in the vitreous within the first two days after treatment and remained at low levels from day 9 onward. The eyes that were injected with acyclovir did not present any electroretinographic changes compared with the control eyes. CONCLUSIONS: The vitreous half-life of acyclovir is short, and the electrophysiological findings suggest that the intravitreal delivery of 1 mg acyclovir is safe and well tolerated by the rabbit retina.

Multifocal electroretinogram in normal emmetropic subjects: Correlation with optical coherence tomography.

Aim of the Study: To establish the normative database for multifocal electroretinogram (mfERG) parameters in a normal emmetropic population. To correlate the data so obtained with the central macular thickness obtained using the optical coherence tomography (OCT) scan. Materials and Methods: mfERG data were obtained from 222 eyes of 111 emmetropic subjects. The amplitude (nv/deg2) and implicit times (ms) of the first-order kernel mfERG responses (N1, P1, and N2 waves) were obtained and grouped into five rings (Ring 1: Central 2°, Ring 2: 2–5°, Ring 3: 5–10°, Ring 4: 10–15°, Ring 5: >15°). The central macular thickness (CMT) was obtained using the macular thickness scan protocol of the OCT. Results: The mfERG data obtained were used to create a normative database. The amplitudes of the mfERG waves were maximum in the fovea and progressively decreased with increasing eccentricity (P = 0.0001). The latencies of the P1 and N2 waves were longest in the central ring and progressively shortened with eccentricity (P = 0.0001). No statistically significant correlations were observed between central ring 1 parameters and the CMT. Conclusion: This study establishes normative database for mfERG parameters in an emmetropic population. No statistically significant correlation was noted between CMT and mfERG parameters.

Axonal electrovisiogram as an electrophysiological test to evaluate optic nerve and inner retina electrical potentials: findings in normal subjects / Eletrovisuograma axonal como teste eletrofisiológico na avaliação dos potenciais de ação do nervo óptico e da retina interna: achados em indivíduos normais

PURPOSES: To standardize and validate the technique of axonal electrovisiogram (AxEvg), defining its normative values and parameters and characterizing its findings in normal individuals. METHODS: We enrolled 140 normal individuals (280 eyes) divided into seven groups according to age, each one with 10 males and 10 females. The technique was based on monocular visual stimulation by a 0 dB intensity bright flash on Ganzfeld bowl at a presentation rate of 1.4 Hz. Golden cup electrodes were used and electrical waves were acquired after artifact rejection. For each amplitude and implicit time peak we calculated the mean, median, pattern deviation, minimum and maximum values and 95 percent confidence interval. RESULTS: Monocular visual stimulation with bright flash under mesopic conditions was the standard technical procedure established. The normal AxEvg waveform consists of an initial positive wave (named P1, with mean amplitude of 2.0 mV and mean implicit time peak of 23.1 ms) followed by a negative wave (named N1, with mean amplitude of -3.9 mV and mean implicit time peak of 41.4 ms). No significant differences were observed between males and females or between right and left eyes, but there was an increased P1 and N1 implicit time peaks according to age. Implicit time characteristics suggest that P1 wave represents an optic nerve electrical potential and N1 wave represents an inner retinal layers potential. CONCLUSIONS: AxEvg can be considered a pre-chiasmatic visual evoked potential capable to reliably record the electrical activity of optic nerve and inner retina. The findings suggest that AxEvg may be useful as an electrophysiological test in the diagnosis of neuroretinal diseases.

OBJETIVOS: Padronizar e validar a técnica de realização do eletrovisuograma axonal (EVA), definir seus valores normativos e caracterizar os achados em indivíduos normais. MÉTODOS: Estudo descritivo com 140 indivíduos (280 olhos) normais. Os participantes foram divididos em sete grupos de acordo com a idade, cada qual com 10 indivíduos do sexo masculino e 10 do sexo feminino. Definiu-se como técnica de exame a estimulação monocular por flash luminoso com intensidade de 0 dB na cúpula de Ganzfeld a uma frequência de 1,4 Hz. Foram utilizados eletrodos com cúpula de ouro e foram analisados os traçados elétricos obtidos após rejeição de artefatos. Para cada amplitude e tempo de culminação foram calculados a média, a mediana, o desvio-padrão, os valores mínimo e máximo e o intervalo de confiança de 95 por cento representando uma faixa de normalidade para os valores. RESULTADOS: A técnica de exame padronizada foi a estimulação visual monocular por flash em condições mesópicas. O traçado normal do eletrovisuograma axonal consistiu numa onda positiva inicial (P1, com amplitude média de 2,0 mV e tempo de culminação médio de 23,1 ms) seguida de uma onda negativa (N1, com amplitude média de -3,9 mV e tempo de culminação médio de 41,4 ms). Não foram observadas diferenças significativas entre os sexos e entre os olhos direito e esquerdo, mas os tempos de culminação de P1 e de N1 aumentaram proporcionalmente com a idade. CONCLUSÕES: O eletrovisuograma axonal é um exame de potencial visual evocado pré-quiasmático, capaz de registrar de forma confiável e reprodutível a atividade elétrica do nervo óptico e da retina interna, podendo ser utilizado na propedêutica eletrofisiológica na investigação de lesões neurorretinianas.

Study for Analysis of the Multifocal Visual Evoked Potential

PURPOSE: To introduce the clinical utility of the absolute value of the reconstructed waveform method in the analysis of multifocal visual evoked potential (mfVEP). METHODS: The mfVEP with 4-channel recording was performed using RETIscan(R) on 10 eyes of 10 normal subjects. Amplitudes were obtained from ring-shaped 6 areas and 4 sectors. The best visual evoked potential (VEP) response method and the absolute value of the reconstructed waveform method were compared in terms of analysis of the amplitudes. In order to assess the false positive rate of the examination, stimuli were administered with one-half of the cathode ray tube (CRT) monitor completely covered and the results were compared using 2 methods. RESULTS: The amplitudes in 6 areas and 4 sectors analyzed with the best VEP response method and the absolute value of the reconstructed waveform method showed no statistical difference (p > 0.05). The amplitude in the stimuli-blocked area of the absolute value of the reconstructed waveform method was smaller than that of the best VEP response method (p 0.05). CONCLUSIONS: The absolute value of the reconstructed waveform method has similar reproducibility and lower level of false positives relative to the best VEP response method. Therefore, it can be considered as a useful method in the analysis of the mfVEP.

Study for Analysis of the Multifocal Visual Evoked Potential

PURPOSE: To introduce the clinical utility of the absolute value of the reconstructed waveform method in the analysis of multifocal visual evoked potential (mfVEP). METHODS: The mfVEP with 4-channel recording was performed using RETIscan(R) on 10 eyes of 10 normal subjects. Amplitudes were obtained from ring-shaped 6 areas and 4 sectors. The best visual evoked potential (VEP) response method and the absolute value of the reconstructed waveform method were compared in terms of analysis of the amplitudes. In order to assess the false positive rate of the examination, stimuli were administered with one-half of the cathode ray tube (CRT) monitor completely covered and the results were compared using 2 methods. RESULTS: The amplitudes in 6 areas and 4 sectors analyzed with the best VEP response method and the absolute value of the reconstructed waveform method showed no statistical difference (p > 0.05). The amplitude in the stimuli-blocked area of the absolute value of the reconstructed waveform method was smaller than that of the best VEP response method (p 0.05). CONCLUSIONS: The absolute value of the reconstructed waveform method has similar reproducibility and lower level of false positives relative to the best VEP response method. Therefore, it can be considered as a useful method in the analysis of the mfVEP.

The synergetic modulation of the excitability of central gray matter by a neuropeptide: two protocols using excitation waves in chick retina

The isolated chick retina provides an in vitro tissue model, in which two protocols were developed to verify the efficacy of a peptide in the excitability control of the central gray matter. In the first, extra-cellular potassium homeostasis is challenged at long intervals and in the second, a wave is trapped in a ring of tissue causing the system to be under self-sustained challenge. Within the neuropil, the extra-cellular potassium transient observed in the first protocol was affected from the initial rising phase to the final concentration at the end of the five-minute pulse. There was no change in the concomitants of excitation waves elicited by the extra-cellular rise of potassium. However, there was an increase on the elicited waves latency and/or a rise in the threshold potassium concentration for these waves to appear. In the second protocol, the wave concomitants and the propagation velocity were affected by the peptide. The results suggest a synergetic action of the peptide on glial and synaptic membranes: by accelerating the glial Na/KATPase and changing the kinetics of the glial potassium channels, with glia tending to accumulate KCl. At the same time, there is an increase in potassium currents through nerve terminals.

Retinas de pinto isoladas proporcionam um modelo de tecidos in vitro, para o qual dois protocolos foram desenvolvidos para verificar a eficácia de um peptídeo no controle da excitabilidade da matéria cinzenta central. No primeiro, a homeostase do potássio extra-celular é desafiada por intervalos longos (1 hora) e no segundo, uma onda é capturada em um anel de tecido, de tal maneira que o sistema permaneça em estado de desafio auto-sustentado. Dentro da neuropil, o transiente de potássio extra-celular observado no primeiro protocolo foi afetado da fase de início de aumento à concentração final, ao final do pulso de cinco minutos. Não há mudanças nos parâmetros concomitantes das ondas de excitação geradas pelo aumento do potássio extra-celular. Entretanto, houve um aumento da latência das ondas geradas e/ou um aumento no nível de concentração de potássio necessário para gerar a onda. No segundo protocolo, os parâmetros concomitantes da onda e sua velocidade de propagação foram afetados pelo peptídeo. Os resultados sugerem uma ação sinergética do peptídeo nas membranas gliais e sinápticas: acelerando o Na/KATPase glial e mudando a cinética dos canais de potássio gliais, com a glia tendendo a acumular KCl. Nesse período, não há aumento nas correntes de potássio nas terminações nervosas.

The PRVEPs evoked by different fields stimulation / 法医学杂志

OBJECTIVE@#To study the character of the PRVEPs evoked by different stimulation fields in normal-vision subjects and its value in forensic medicine.@*METHODS@#The PRVEPs were evoked by full-field, central-field and peripheral-field respectively in 30 subjects (60 eyes) and the latency and amplitude of N1 and P1 were analyzed.@*RESULTS@#There was no statistically significant difference in the latency and amplitude of PRVEPs between right and left eyes. The appearance probability of all the waves was 100% at the different stimulations but that of N1 was 36.77% at the central-field. There was statistically significant difference in the latency of N1 at the full field and peripheral field stimulations and there was no statistically significant difference in the amplitude. There was statistically significant difference in the latency and amplitude of P1 at the full field, central field and peripheral field stimulation.@*CONCLUSION@#The PRVEPs evoked by different field stimulations can be used to evaluate the functions of the different area retina and can be used to identify the exaggerated or pretended visual dysfunction.

The electroretinogram as a method for studying circadian rhythms in the mammalian retina.

Circadian clocks are thought to regulate retinal physiology in anticipation of the large variation in environmental irradiance associated with the earth's rotation upon its axis. In this review we discuss some of the rhythmic events that occur in the mammalian retina, and their consequences for retinal physiology. We also review methods of tracing retinal rhythmicity in vivo and highlight the electroretinogram (ERG) as a useful technique in this field. Principally, we discuss how this technique can be used as a quick and noninvasive way of assessing physiological changes that occur in the retina over the course of the day. We highlight some important recent findings facilitated by this approach and discuss its strengths and limitations.

Stem cells: potential source for retinal repair and regeneration

Stem cells have been studied in several fields of Medicine, and their applications are not too far from the clinical practice. Retinal impairment by neuronal death has been considered incurable due to the limited regenerative capacity of the central nervous system. The capacity of stem cells to regenerate tissues, as well as their plasticity makes them a potential source for retinal repair. The stem cells are a great promise for the therapy of inherited retinal disorders and retinal-neuronal degenerative diseases, such as retinitis pigmentosa and allied retinal dystrophies, which can result in blindness. Because of the accessibility, expansibility, and multipotentiality mesenchymal stem cells are expected to be useful for clinical applications, especially in regenerative medicine and tissue engineering. Mesenchymal stem cells are clonogenic, nonhematopoietic stem cells present in the bone marrow. Given the appropriate microenvironment, they could differentiate into cardiomyocytes or even into cells of nonmesodermal derivation including hepatocytes and neurons. So far, the results of a few studies are consistent with the belief that cell-based therapies using mesenchymal stem cells may be effective when it comes to retinal damaged tissue repair.

Células-tronco têm sido estudadas em várias áreas da Medicina e suas aplicações brevemente deverão estar incorporadas à prática clínica. O dano retiniano pela morte neuronal é considerado incurável devido a pobre capacidade regenerativa do sistema nervoso central. A capacidade das células-tronco em regenerar tecidos, assim como sua plasticidade, faz que estas sejam uma fonte potencial de células para a regeneração retiniana. Células-tronco são muito promissoras para o tratamento das distrofias retinianas, como a retinose pigmentar e outras doenças neurodegenerativas, que podem evoluir para cegueira. As células-tronco mesenquimais são o tipo mais provável de células-tronco a serem utilizadas na prática clínica devido a sua fácil acessibilidade e multipotencialidade de diferenciação em vários tecidos. As células-tronco mesenquimais são células clonogênicas, não-hematopoiéticas, localizadas na medula óssea. Desde que seja proporcionado um microambiente apropriado, estas células podem se diferenciar em cardiomiócitos e até mesmo em células de origem não-mesodérmica, como hepatócitos e neurônios. Até o presente momento, os resultados dos estudos iniciais são animadores em relação ao uso de células-tronco mesenquimais e uso eficaz destas no reparo de tecidos retinianos lesados.

The determination of dark adaptation time using electroretinography in conscious Miniature Schnauzer dogs

The optimal dark adaptation time of electroretinograms (ERG's) performed on conscious dogs were determined using a commercially available ERG unit with a contact lens electrode and a built-in light source (LED-electrode). The ERG recordings were performed on nine healthy Miniature Schnauzer dogs. The bilateral ERG's at seven different dark adaptation times at an intensity of 2.5 cd.s/m2 was performed. Signal averaging (4 flashes of light stimuli) was adopted to reduce electrophysiologic noise. As the dark adaptation time increased, a significant increase in the mean a-wave amplitudes was observed in comparison to base-line levels up to 10 min (p > 0.05). Thereafter, no significant differences in amplitude occured over the dark adaptation time. Moreover, at this time the mean amplitude was 60.30 +/- 18.47 microV. However, no significant changes were observed for the implicit times of the a-wave. The implicit times and amplitude of the b-wave increased significantly up to 20 min of dark adaptation (p > 0.05). Beyond this time, the mean b-wave amplitudes was 132.92 +/- 17.79 microV. The results of the present study demonstrate that, the optimal dark adaptation time when performing ERG's, should be at least 20 min in conscious Miniature Schnauzer dogs.

Visual field (Octopus 1-2-3) in normal subjects divided into homogeneous age-groups / Perimetria computadorizada no Octopus 1-2-3: estudo de uma população normal por faixas etárias estratificadas

PURPOSE: To determine the values in decibels of retinal sensitivity within the central 26 degrees of the visual field of normal subjects divided into homogenous age groups using the Octopus 1-2-3; to compare the values of retinal sensitivity we have found with those considered normal in the statistical package obtained by a multicenter study performed in 1994 with Octopus 201. METHODS: 181 subjects divided into 6 homogeneous age groups (10 to 19 yr; 20 to 29 yr; 30 to 39 yr; 40 to 49 yr; 50 to 59 yr and 60 year-old or older) were evaluated. Data on visual sensitivity and age, average sensitivity of central and paracentral regions and eccentricity were calculated. RESULTS: The average visual sensitivity of all groups was 26.77 ± 1.74 dB. Correlation between visual sensitivity and age evaluated by linear regression was 28.4 - 0.040 x (age) for the whole sample and 28.7 - 0.050 x (age) for subjects aged 20 or more. Sensitivity reduction by eccentricity was -0.30 dB/degree for the whole sample and for subjects aged 20 or more. CONCLUSIONS: Correlation between retinal sensitivity values and age based on the autoperimeter Octopus 201 (average sensitivity of 31.2 - 0.064 x age) is different from that found in this study: average sensitivity of 28.4 - 0.040 x (age) for the whole sample; 28.7 - 0.050 x (age) for subjects aged 20 or more. Values obtained with the Octopus 1-2-3 autoperimeter cannot be compared with those by other Octopus models (101, 201 and 500) due to their distinct features.

OBJETIVO: Determinar, utilizando o autoperímetro Octopus 1-2-3, os valores da sensibilidade retiniana em dB, nos 26 graus centrais do campo visual, em voluntários normais, distribuídos em grupos etários homogêneos. Comparar os valores da sensibilidade retiniana com aqueles considerados normais no pacote estatístico do programa do autoperímetro Octopus 1-2-3 obtidos por estudo multicêntrico realizado em 1994. MÉTODOS: Avaliaram-se 181 voluntários, distribuídos em seis grupos etários homogêneos: 10 a 19; 20 a 29; 30 a 39; 40 a 49; 50 a 59 e 60 e mais anos. Foram calculados dados relativos à sensibilidade visual e idade, sensibilidade média nas regiões central e paracentral e influência da excentricidade. RESULTADOS: A sensibilidade visual média de todos os grupos foi de 26,77 dB (desvio-padrão de 1,74 dB). A relação entre sensibilidade visual e idade avaliada pela regressão linear foi de 28,4 - 0,04 x (idade) para toda a amostra e de 28,7 - 0,05 x (idade) para maiores de 19 anos. A redução da sensibilidade com a excentricidade foi de -0,30 dB/grau para toda a amostra e de -0,30 dB/grau para maiores de 19 anos. CONCLUSÕES: Os valores da sensibilidade retiniana encontrados neste estudo: 28,4 - 0,04 x (idade) para toda a amostra e 28,7 - 0,05 x (idade) para maiores de 19 anos são diferentes daqueles utilizados nos autoperímetros Octopus e, a rigor, não podem ser comparados aos obtidos nos outros modelos Octopus (101, 201 e 500) em face das suas distintas características.

Study of Blue and Red Flash in Dark-Adapted Electroretinogram

PURPOSE: To investigate the characteristics of the waveform generated by blue and red light stimulations in a dark-adapted electroretinogram (ERG) and those of cone responses in the dark-adapted condition. METHODS: The study subjects were 52 persons (88 eyes) with no previous medical history. The author recorded b-waves (rod response) with red light stimulation and the x-waves (dark-adapted cone response) that appeared before the b-waves. The author also recorded b-waves with blue light stimulation, which had the same amplitude as the b-waves from the red light stimulation. The differences with respect to age and gender were studied. Waveforms of the dark-adapted cone ERGs were recorded by using a digital subtraction technique. RESULTS: The x-wave always appeared before the b-wave with 0 dB (2.4 cd.s/m2) red stimulation. With blue stimulation, a b-wave equivalent to the b-wave stimulated with the red light of 0 dB intensity appeared at an average of -14.57 dB. The implicit time for the b-wave was delayed significantly for the male group. There were no significant differences between different age groups. The dark-adapted cone ERG demonstrated the waveform of a negative response followed by a series of oscillatory potentials (OPs) and a positive response. CONCLUSIONS: The cone responses were followed by the rod responses with red light stimulation of 0 dB in the dark-adapted ERG. The waveforms of the cone ERGs were obtained in dark adaptation with red and blue light stimulation.

Liquid crystal display microperimetry in eyes with reduced visual acuity from macular pathology.

PURPOSE: To correlate fixation stability and retinal sensitivity measured by liquid crystal display (LCD) microperimetry to visual acuity in eyes with macular pathology. DESIGN: Retrospective chart review METHODS: The cohort included 26 eyes of 26 patients, divided into two groups according to visual acuity (Group 1 eyes 6/15 - 6/48, Group 2: < or = 6/60). Macular sensitivity and fixation stability were measured using the LCD microperimeter. Mean retinal sensitivity (at central 12 degrees) and fixation stability (at 2 degrees and 4 degrees) were correlated with best-corrected visual acuity. RESULTS: Mean retinal sensitivity correlated with the visual acuity in both groups [Group 1 eyes: 28 point central 12 degrees (9.0 dB); Group 2: central 12 degrees (4.183 dB)] (P < 0.05). The fixation stability correlated similarly with the visual acuity for the groups [Group 1 eyes: 2 degrees (81.2%) and 4 degrees (94.6%); Group 2 eyes: 2 degrees (44.1%) and 4 degrees (80.7%] (P < 0.05). CONCLUSION: In the present study, the mean retinal sensitivity and fixation stability measured with LCD microperimeter correlated with visual acuity. LCD microperimetry reliably measures fixation stability and retinal sensitivity in eyes with various macular pathologies.

Wave onset in central gray matter - its intrinsic optical signal and phase transitions in extracellular polymers

The brain is an excitable media in which excitation waves propagate at several scales of time and space. ''One-dimensional'' action potentials (millisecond scale) along the axon membrane, and spreading depression waves (seconds to minutes) at the three dimensions of the gray matter neuropil (complex of interacting membranes) are examples of excitation waves. In the retina, excitation waves have a prominent intrinsic optical signal (IOS). This optical signal is created by light scatter and has different components at the red and blue end of the spectrum. We could observe the wave onset in the retina, and measure the optical changes at the critical transition from quiescence to propagating wave. The results demonstrated the presence of fluctuations preceding propagation and suggested a phase transition. We have interpreted these results based on an extrapolation from Tasaki's experiments with action potentials and volume phase transitions of polymers. Thus, the scatter of red light appeared to be a volume phase transition in the extracellular matrix that was caused by the interactions between the cellular membrane cell coat and the extracellular sugar and protein complexes. If this hypothesis were correct, then forcing extracellular current flow should create a similar signal in another tissue, provided that this tissue was also transparent to light and with a similarly narrow extracellular space. This control tissue exists and it is the crystalline lens. We performed the experiments and confirmed the optical changes. Phase transitions in the extracellular polymers could be an important part of the long-range correlations found during wave propagation in central nervous tissue