A tubulização representa uma possível alternativa para reparo de lesões de nervos periféricos, por permitir a aplicação local de fatores de crescimento e neurorregenerativos assim como por facilitar o estudo dos mecanismos de ação relacionados à regeneração das fibras nervosas. O objetivo do presente estudo foi verificar se há crescimento de fibras amielínicas no nervo fibular comum de ratas Wistar adultas após a tubulização látero-terminal (TLT) sem a utilização de indutores de crescimento, através de análise morfológica e morfométrica das referidas fibras. Para tanto, o nervo fibular comum direito foi seccionado a 3 mm da sua emergência, o coto proximal sepultado e suturado na musculatura adjacente e um tubo de silicone (6 mm) interposto entre o coto distal do fibular e a lateral do nervo tibial (grupo TLT). O controle usado foi o segmento distal esquerdo do nervo fibular (grupo GN). Setenta dias após, foram realizados cortes semifinos e ultrafinos para análise de aspectos morfológicos e morfométricos através de microscopia de luz e microscopia eletrônica de transmissão (MET). Houve evidente brotamento axonal de fibras amielínicas, que cresceram a partir do nervo tibial intacto no grupo TLT em comparação ao grupo GN, sendo que o número total de axônios amielínicos foi similar nos dois grupos. Estudo morfométrico evidenciou diferenças significativas (p<=0,05) em relação à maior densidade de axônios amielínicos regenerados e em relação ao diâmetro mínimo axonal. No presente estudo os axônios amielínicos no nervo receptor, usando o modelo da TLT, foram quantificados. Esse modelo poderá ser útil e importante para estudo da plasticidade do sistema nervoso periférico. / Side-to-end tubulisation is a model recently developed in our laboratory to study nerve plasticity. In this model, collateral sprouting of fibers grow from intact donor nerve fibers to the distal stump of a receptor nerve. The objective of the present study was to study unmyelinated fibers using this model. Adults female Wistar rats were used and morphological and morphometric analysis were done. The fibular common nerve was sectioned 3 mm from its origin. The proximal stump was buried and sutured in the adjacent musculature. A silicone tube (6 mm) was fixed on the adjacent lateral portion of the intact tibial nerve and the fibular common nerve distal stump was sutured in the other extremity of the tube (SET group). The left fibular common nerve distal segment was used as control. Semithin and ultrathin sections were obtained and studied using light and transmission electron microscopy. Seventy days after, there was profuse axonal sprouting of unmyelinated fibers that grew from the nerve tibial, reaching the same number of axons when compared with the controls. In the present study unmyelinated axons in the receptor nerve using SET were quantified. SET is an important tool to study plasticity of peripheral nerve fibers.

No description available.

Abstract The aim of this study was to create a mathematical model of the retinal nerve fiber layer and of the entire hill of vision, and to compare different perimetric methods and test grids in the detection of visual field loss in glaucoma and optic neuritis. A mathematical model of the retinal nerve fiber layer was developed, based on traced nerve fiber bundle trajectories extracted from 55 fundus photographs of 55 human subjects. The model resembled the typical retinal nerve fiber layer course within 20° eccentricity from the foveola. The standard deviation of the calculated corresponding angular location at the optic nerve head circumference ranged from less than 1° up to 18° (mean 8.8°). A smooth mathematical model of the hill of vision was created, based on 81 ophthalmologically healthy subjects. The model fit R2 was 0.72. Applying individually condensed test grids in 41 glaucomatous eyes of 41 patients enhanced remarkably the detection of progression. Seven out of 11 (64%) of the progressive scotomata detected by spatially condensed grids would have been missed by the conventional 6° × 6° grid. In 20 eyes of 20 patients with advanced glaucoma, the comparability of visual field areas obtained with semi-automated kinetic perimetry and automated static perimetry was satisfactory and within the range of the test-retest reliability of automated static perimetry. Using a standardized grid of 191 static targets within the central 30° visual field, the most common finding in 100 eyes of 99 patients with acute optic neuritis were central scotomas, accounting for 41% of all visual field defects in affected eyes. In conclusion, a model of the retinal nerve fiber layer was developed, which provided a detailed location specific estimate of the magnitude of the variability on the courses of retinal nerve fiber bundle trajectories in the human retina. A smooth mathematical model of the hill of vision with a satisfactory model fit was described for the 80° visual field. Individually condensed grids enabled the detection of a glaucomatous visual field progression more frequently and also earlier than conventional grids. Semi-automated kinetic perimetry was found to be a valuable alternative to automated static perimetry in monitoring advanced glaucomatous visual field loss. Using a grid with a higher spatial resolution may enhance the detection of small central visual field loss in optic neuritis.

博士(理学) / Doctor of Philosophy in Science / 同志社大学 / Doshisha University

The moment-to-moment fluctuations in pain-evoked sensory and emotional qualities, and how the pain experience differs between sexes are not well understood. Therefore, this thesis sought to 1) characterise the temporal profiles of the most prominent noxious heat-evoked sensations, 2) characterise sex differences in these sensations, 3) evaluate the magnitude of sharp pain quality evoked in hairy and glabrous skin, and 4) determine the role of absolute stimulus temperatures on sex differences in pain adaptation and habituation. A broad-based heat pain model was developed for this study that incorporates a temporally-continuous assessment of multiple sensory and affective pain dimensions, including pain, burning, sharp, stinging, cutting, and annoyance evoked by two types (static, dynamic) of repeated prolonged noxious heat stimuli. The salient hypotheses were: 1) Burning sensations have a different temporal profile compared with sharp and other related qualities, 2) The temporal dynamics of heat pain intensity and annoyance differ between males and females, 3) Sex differences in heat pain are associated with specific pain qualities and specific types of skin, and 4) Moderate-high temperatures induce pain adaptation and habituation in females but not in males. The most prominent findings were 1) sharp, stinging and cutting sensations adapted when stimulus intensity was static, but burning sensations were evoked during static and dynamic stimulus phases, 2) pain and annoyance in women were greater than men during the dynamic phases of the first stimulus but less than men during static stimulus phases and on stimulus repetition, 3) the sex difference in pain adaptation occurred with percept-fixed stimulus intensities and with absolute stimulus temperatures, 4) the sex effects associated with dynamic stimuli occurred in hairy but not glabrous skin. These findings give new insights into the relationships between pain intensity, quality and affect and have strong implications for views on sex differences in pain sensitivity.

The moment-to-moment fluctuations in pain-evoked sensory and emotional qualities, and how the pain experience differs between sexes are not well understood. Therefore, this thesis sought to 1) characterise the temporal profiles of the most prominent noxious heat-evoked sensations, 2) characterise sex differences in these sensations, 3) evaluate the magnitude of sharp pain quality evoked in hairy and glabrous skin, and 4) determine the role of absolute stimulus temperatures on sex differences in pain adaptation and habituation. A broad-based heat pain model was developed for this study that incorporates a temporally-continuous assessment of multiple sensory and affective pain dimensions, including pain, burning, sharp, stinging, cutting, and annoyance evoked by two types (static, dynamic) of repeated prolonged noxious heat stimuli. The salient hypotheses were: 1) Burning sensations have a different temporal profile compared with sharp and other related qualities, 2) The temporal dynamics of heat pain intensity and annoyance differ between males and females, 3) Sex differences in heat pain are associated with specific pain qualities and specific types of skin, and 4) Moderate-high temperatures induce pain adaptation and habituation in females but not in males. The most prominent findings were 1) sharp, stinging and cutting sensations adapted when stimulus intensity was static, but burning sensations were evoked during static and dynamic stimulus phases, 2) pain and annoyance in women were greater than men during the dynamic phases of the first stimulus but less than men during static stimulus phases and on stimulus repetition, 3) the sex difference in pain adaptation occurred with percept-fixed stimulus intensities and with absolute stimulus temperatures, 4) the sex effects associated with dynamic stimuli occurred in hairy but not glabrous skin. These findings give new insights into the relationships between pain intensity, quality and affect and have strong implications for views on sex differences in pain sensitivity.

INTRODUÇÃO: A toxina botulínica tipo A (TXB-A) tem sido testada no tratamento da rinite, principalmente nos casos de rinite idiopática. Sugere-se que um estado de hiper-reatividade do nervo trigêmeo esteja envolvido na fisiopatologia da rinite idiopática. O nervo trigêmeo possui fibras sensitivas não mielinizadas tipo C (FSNMT-C) que contém os neuropeptídeos substância P (SP) e peptídeo relacionado ao gene da calcitonina (CGRP). O óxido nítrico (NO) produzido pelas enzimas óxido nítrico sintase (NOS) também está envolvido nesse processo de neurorregulação nasal. O transporte mucociliar, mecanismo primário de defesa do sistema respiratório, é formado pelo batimento ciliar e muco nasal, e esses componentes podem ser influenciados por diferentes neuropeptídeos e neurotransmissores presentes na mucosa nasal. OBJETIVO: O objetivo deste estudo foi avaliar o efeito da TXB-A sobre a expressão da SP, CGRP e óxido nítrico sintase neural (nNOS), além de sua influência sobre o transporte mucociliar nasal em coelhos. MÉTODOS: Coelhos machos saudáveis da linhagem Nova Zelândia foram divididos em dois grupos: o grupo tratamento recebeu TXB-A (25UI) na concha nasomaxilar (CNM) do lado direito e soro fisiológico a 0,9% (SF0,9%) na CNM esquerda. O grupo controle recebeu SF0,9% na CNM direita e nenhuma intervenção na CNM esquerda. Foram investigados os efeitos da TXB-A sobre a expressão da SP, CGRP e nNOS no tecido de CNM por meio da imuno-histoquímica. Para esta análise, dividiu-se o tecido em camada externa (CE, acima da membrana basal) e camada interna (CI, abaixo da membrana basal). Avaliou-se também a presença de apoptose celular, a frequência de batimento ciliar (FBC), o perfil histoquímico do muco nasal (glicoproteínas ácidas e neutras) e a espessura do epitélio (ESP-CE). RESULTADOS: Foi observado um aumento significativo na quantidade de células apoptóticas na CNM do grupo tratamento que recebeu TXB-A em comparação aos controles (p <= 0,001). A CNM do grupo tratamento que recebeu SF0,9% exibiu um aumento na quantidade de células apoptóticas na CI ao comparar com os controles (CNM SF0,9%, p=0,035) (CNM sem intervenção, p=0,022), e também um aumento da expressão de SP na CE em comparação aos controles (CNM SF0,9%, p=0,021) (CNM sem intervenção, p=0,040). A expressão de CGRP apresentou um aumento na CNM do grupo tratamento que recebeu TXB-A em comparação à CNM sem intervenção (p=0,008). A FBC, expressão de nNOS, perfil histoquímico do muco nasal e ESP-CE não apresentaram diferenças significativas. DISCUSSÃO: O aumento da expressão de CGRP e SP pode ter sido causado por inibição de sua exocitose vesicular pela TXB-A, levando ao seu acúmulo intracelular. Não foram observadas diferenças significativas na FBC ou perfil histoquímico do muco nasal, indicando que o aumento da expressão de CGRP e SP não foi associado à inflamação. O aumento da quantidade de células apoptóticas e da expressão de SP na CNM SF0,9% do grupo tratamento pode ter sido causado por um efeito central da TXB-A. CONCLUSÃO: A administração nasal de TXB-A aumentou a expressão de CGRP e SP, possivelmente por acúmulo intracelular por causa da inibição da sua exocitose, mas sem alterar a FBC e o perfil histoquímico do muco nasal / INTRODUCTION: Botulinum toxin type A (BoNT-A) has been assessed in the treatment of rhinitis, especially in cases of idiopathic rhinitis. Trigeminal hyper-responsiveness appears to be involved in the pathological process of idiopathic rhinitis. Trigeminal nociceptive type C unmyelinated sensory fibers contain the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). Nitric oxide (NO) produced by the enzyme nitric oxide synthase (NOS) are also involved in this nasal neurorregulation process. The mucociliary clearance, primary defense system of the respiratory system, is composed by the ciliary beat and nasal mucus. These components can be influenced by different nasal neuropeptides and neurotransmitters. OBJECTIVE: The aim of this study was to evaluate the effect of BoNT-A on the expression of SP, CGRP and neural nitric oxide synthase (nNOS), and its influence on nasal mucociliary clearance in rabbits. METHODS: Healthy New Zealand male rabbits were divided into two groups: the treatment group was challenged with BoNT-A (25UI) in the right nasomaxillary turbinate (NMT) and saline (SF0.9%) in the left NMT. The control group received SF0.9% in the right NMT and no-intervention in the left NMT. We investigated the effects of BoNT-A on SP, CGRP and nNOS expression in the NMT tissue by immunohistochemistry. Each area of interest was subdivided into an internal layer (IL: below the basement membrane) and outer layer (OL: above the basement membrane) for analysis. It was also assessed signs of cellular apoptosis, ciliary beat frequency (CBF), mucus histochemical profile (acidic and neutral glycoproteins) and epithelial thickness (EP-TH). RESULTS: It was observed a significant increase in the amount of apoptotic cells in the BoNT-A-challanged NMT compared with controls (p <= 0.001). The NMT of treatment group which received only SF0.9% showed an increase in the amount of apoptotic cells in the IL compared with controls (NMT SF0.9%, p = 0.035) (NMT no-intervention, p = 0.022), and also an increase in the SP expression in the OL compared with controls (NMT SF0.9%, p = 0.021) (NMT no-intervention, p = 0.040). CGRP expression showed higher expression in the BoNT-A-challanged NMT compared with no-intervention NMT (p=0.008). The CBF, nNOS expression, mucus histochemical profile and EP-TH did not show significant differences. DISCUSSION: The increased CGRP and SP expression could be due to inhibition of vesicular exocytosis by BoNT-A, leading to CGRP and SP intracellular accumulation. No significant differences in CBF or mucus histochemical profile were observed, indicating that the increased CGRP and SP expression was not associated with inflammation. The increase in the amount of apoptotic cells and SP expression in the SF0.9% NMT of treatment group may be due to a central effect of BoNT-A. CONCLUSION: Nasal administration of BoNT-A increased SP and CGRP expression, possibly via inhibition of their release, but did not change the CBF or mucus profile

INTRODUÇÃO: A toxina botulínica tipo A (TXB-A) tem sido testada no tratamento da rinite, principalmente nos casos de rinite idiopática. Sugere-se que um estado de hiper-reatividade do nervo trigêmeo esteja envolvido na fisiopatologia da rinite idiopática. O nervo trigêmeo possui fibras sensitivas não mielinizadas tipo C (FSNMT-C) que contém os neuropeptídeos substância P (SP) e peptídeo relacionado ao gene da calcitonina (CGRP). O óxido nítrico (NO) produzido pelas enzimas óxido nítrico sintase (NOS) também está envolvido nesse processo de neurorregulação nasal. O transporte mucociliar, mecanismo primário de defesa do sistema respiratório, é formado pelo batimento ciliar e muco nasal, e esses componentes podem ser influenciados por diferentes neuropeptídeos e neurotransmissores presentes na mucosa nasal. OBJETIVO: O objetivo deste estudo foi avaliar o efeito da TXB-A sobre a expressão da SP, CGRP e óxido nítrico sintase neural (nNOS), além de sua influência sobre o transporte mucociliar nasal em coelhos. MÉTODOS: Coelhos machos saudáveis da linhagem Nova Zelândia foram divididos em dois grupos: o grupo tratamento recebeu TXB-A (25UI) na concha nasomaxilar (CNM) do lado direito e soro fisiológico a 0,9% (SF0,9%) na CNM esquerda. O grupo controle recebeu SF0,9% na CNM direita e nenhuma intervenção na CNM esquerda. Foram investigados os efeitos da TXB-A sobre a expressão da SP, CGRP e nNOS no tecido de CNM por meio da imuno-histoquímica. Para esta análise, dividiu-se o tecido em camada externa (CE, acima da membrana basal) e camada interna (CI, abaixo da membrana basal). Avaliou-se também a presença de apoptose celular, a frequência de batimento ciliar (FBC), o perfil histoquímico do muco nasal (glicoproteínas ácidas e neutras) e a espessura do epitélio (ESP-CE). RESULTADOS: Foi observado um aumento significativo na quantidade de células apoptóticas na CNM do grupo tratamento que recebeu TXB-A em comparação aos controles (p <= 0,001). A CNM do grupo tratamento que recebeu SF0,9% exibiu um aumento na quantidade de células apoptóticas na CI ao comparar com os controles (CNM SF0,9%, p=0,035) (CNM sem intervenção, p=0,022), e também um aumento da expressão de SP na CE em comparação aos controles (CNM SF0,9%, p=0,021) (CNM sem intervenção, p=0,040). A expressão de CGRP apresentou um aumento na CNM do grupo tratamento que recebeu TXB-A em comparação à CNM sem intervenção (p=0,008). A FBC, expressão de nNOS, perfil histoquímico do muco nasal e ESP-CE não apresentaram diferenças significativas. DISCUSSÃO: O aumento da expressão de CGRP e SP pode ter sido causado por inibição de sua exocitose vesicular pela TXB-A, levando ao seu acúmulo intracelular. Não foram observadas diferenças significativas na FBC ou perfil histoquímico do muco nasal, indicando que o aumento da expressão de CGRP e SP não foi associado à inflamação. O aumento da quantidade de células apoptóticas e da expressão de SP na CNM SF0,9% do grupo tratamento pode ter sido causado por um efeito central da TXB-A. CONCLUSÃO: A administração nasal de TXB-A aumentou a expressão de CGRP e SP, possivelmente por acúmulo intracelular por causa da inibição da sua exocitose, mas sem alterar a FBC e o perfil histoquímico do muco nasal / INTRODUCTION: Botulinum toxin type A (BoNT-A) has been assessed in the treatment of rhinitis, especially in cases of idiopathic rhinitis. Trigeminal hyper-responsiveness appears to be involved in the pathological process of idiopathic rhinitis. Trigeminal nociceptive type C unmyelinated sensory fibers contain the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). Nitric oxide (NO) produced by the enzyme nitric oxide synthase (NOS) are also involved in this nasal neurorregulation process. The mucociliary clearance, primary defense system of the respiratory system, is composed by the ciliary beat and nasal mucus. These components can be influenced by different nasal neuropeptides and neurotransmitters. OBJECTIVE: The aim of this study was to evaluate the effect of BoNT-A on the expression of SP, CGRP and neural nitric oxide synthase (nNOS), and its influence on nasal mucociliary clearance in rabbits. METHODS: Healthy New Zealand male rabbits were divided into two groups: the treatment group was challenged with BoNT-A (25UI) in the right nasomaxillary turbinate (NMT) and saline (SF0.9%) in the left NMT. The control group received SF0.9% in the right NMT and no-intervention in the left NMT. We investigated the effects of BoNT-A on SP, CGRP and nNOS expression in the NMT tissue by immunohistochemistry. Each area of interest was subdivided into an internal layer (IL: below the basement membrane) and outer layer (OL: above the basement membrane) for analysis. It was also assessed signs of cellular apoptosis, ciliary beat frequency (CBF), mucus histochemical profile (acidic and neutral glycoproteins) and epithelial thickness (EP-TH). RESULTS: It was observed a significant increase in the amount of apoptotic cells in the BoNT-A-challanged NMT compared with controls (p <= 0.001). The NMT of treatment group which received only SF0.9% showed an increase in the amount of apoptotic cells in the IL compared with controls (NMT SF0.9%, p = 0.035) (NMT no-intervention, p = 0.022), and also an increase in the SP expression in the OL compared with controls (NMT SF0.9%, p = 0.021) (NMT no-intervention, p = 0.040). CGRP expression showed higher expression in the BoNT-A-challanged NMT compared with no-intervention NMT (p=0.008). The CBF, nNOS expression, mucus histochemical profile and EP-TH did not show significant differences. DISCUSSION: The increased CGRP and SP expression could be due to inhibition of vesicular exocytosis by BoNT-A, leading to CGRP and SP intracellular accumulation. No significant differences in CBF or mucus histochemical profile were observed, indicating that the increased CGRP and SP expression was not associated with inflammation. The increase in the amount of apoptotic cells and SP expression in the SF0.9% NMT of treatment group may be due to a central effect of BoNT-A. CONCLUSION: Nasal administration of BoNT-A increased SP and CGRP expression, possibly via inhibition of their release, but did not change the CBF or mucus profile