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Age-related changes in end-plate architectureRosenheimer, Julie Louise. January 1984 (has links)
Thesis (Ph. D.)--University of Wisconsin--Madison, 1984. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 99-115).
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Anatomy of the nodose ganglion in the rat: central projections of afferent fibers in the hepatic vagus.Pipkin, Bonnie E. 01 January 1983 (has links) (PDF)
Horseradish peroxidase (HRP) was applied to the rostral end of sectioned hepatic vagi (HV) . Subsequently, a count of HRP—labeled cells in the nodose ganglia (NDG) yielded an estimate of the minimum number of afferent fibers in the HV of 139. HHP labeled cells were found only in the left NDG and were diffusely spread throughout the ganglia. No HRP labeling was found in areas of the brain previously reported to contain vagal afferent projections. In three cases small numbers of HRP labeled cell bodies were seen in the dorsal motor nucleus (DMN) . The NDG were organized with cell bodies on the sides and their processes and fibers of passage in the center. The NDG have an apparent population of two cell types; large sensory neurons and smaller glial cells. However, the possibility of a population of smaller sensory cells is discussed. An average of total sensory cell counts for three NDG yielded an estimate of 9115 sensory cells.
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Protection of neuromuscular sensory endings by the WldS geneOyebode, Oyinlola R. O. January 2009 (has links)
The compartmental hypothesis of neurodegeneration proposes that the neurone, long recognized to consist of morphologically and functionally distinct compartments, also houses distinct degeneration mechanisms for the soma, axon and nerve endings. Support for this hypothesis is provided by the phenomenon of the WldS (for Wallerian Degeneration, slow) mouse, a mutant in which axons survive several weeks after transection, rather than degenerating within 24-48 hours as in wild type mice, by virtue of expression of a chimeric Nmnat1/Ube4b protein. In this thesis I used the WldS-mouse to re-examine and extend the theory of compartmental neurodegeneration by focusing specifically on sensory axons and endings; and finally by considering a fourth compartment, the dendrites. The first part of this thesis reports that Ia afferent axons and their annulospiral endings are robustly protected from degeneration in WldS mice. Homozygous or heterozygous WldS mice crossbred with transgenic mice expressing fluorescent protein in neurones were sacrificed at various times after sciatic nerve transection. Fluorescence microscopy of whole mount preparations of lumbrical muscles in these mice revealed excellent preservation of annulospiral endings on muscle spindles for at least 10 days after axotomy. No significant difference was detected in the protection with age or gene copy-number in contrast to the protection of motor nerve terminals, which degenerate rapidly in heterozygote and aged homozygote WldS mice. In an attempt to explain the difference in motor and sensory protection by WldS, examination of three hypotheses was undertaken: a) differences in protein expression, tested by western blot and immunohistochemistry; b) differences in the degree of neuronal branching, tested through examination of g-motor axons and endings which have a degree of branching intermediate to motor and sensory neurons; and c) differences in the activity in the disconnected stumps, through primary culture of the saphenous and phrenic nerve, selected because they comprise largely pure sensory and motor axons respectively. The data suggest that none of these hypotheses provides a sufficient explanation for the difference between sensory and motor protection by WldS. The last part of this thesis attempts to extend the theory of compartmental degeneration. I examine a system for investigation of WldS-mediated protection of dendrites. In preliminary experiments retinal explants from transgenic mice expressing YFP in a subset of retinal ganglion-cell neurones were cultured. The dendritic arbours of these cells were shown to be amenable for repeated visualization and accessible to injury and monitoring of degeneration. Overall the data in this thesis suggest that the level of WldS -mediated protection conferred to an axon or axonal endings varies between different neuronal types. This has implications for the potential applications of WldS research to clinical problems. Specifically, the data imply that sensory neuropathies may benefit more than motor neuropathies from treatments based on the protective effects of WldS. These findings in sensory neurones also challenge some of the assumptions made about WldS- mediated protection of neurones, for example the extent of the age-effect on axonal endings. Further investigation of WldS-mediated protection in the CNS could give renewed impetus to attempts to discover targets for treatment in common neurodegenerative diseases. Finally, a system for investigation of dendritic degeneration has been piloted, suggesting that molecules involved in the degeneration of dendrites or in protection from this degeneration may be amenable to investigation in this system, prospectively extending the compartmental hypothesis of neuronal degeneration.
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Nerve terminal protein complexes in the cholinergic synapse /Sunderland, William James, January 1998 (has links)
Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [104]-122).
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Histomorfometria dos mecanorreceptores e terminaÃÃes nervosas livres no quadril artrÃsico: estudo comparativo com quadril normal de cadÃver / Histomorphometry of mechanoreceptors and free nerve endings in hip joint: a comparative study in patients with secondary hip arthrosis and normal.Miguel Ricardo Barbosa Moraes 16 December 2008 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O interesse de pesquisadores pelo estudo do sistema proprioceptivo vem crescendo nas Ãltimas dÃcadas. Isto à decorrente da importÃncia da integridade deste sistema no tratamento das enfermidades ortopÃdicas. Os mecanorreceptores e as terminaÃÃes nervosas livres sÃo as unidades microscÃpicas fundamentais da propriocepÃÃo e do sistema nervoso aferente. Eles transmitem ao sistema nervoso central informaÃÃes fisiolÃgicas em forma de potencial de aÃÃo, quando as estruturas articulares, cÃpsula, ligamentos e mÃsculos, sÃo submetidas ao estresse do movimento. Foram avaliadas a presenÃa e a densidade dos mecanorreceptores na cÃpsula, ligamento da cabeÃa femoral e labrum acetabular de 45 quadris masculinos. Destes, 30 foram obtidos de pacientes do sexo masculino com artrose secundÃria (grupo I) durante artroplastia e 15 de cadÃveres frescos com articulaÃÃo do quadril normal (grupo II). A idade mÃdia do grupo I foi de 56,5 e do grupo II foi de 35,6 anos. Os fragmentos obtidos foram mensurados em 2mm e corados com cloreto de ouro a 1%. ApÃs fixaÃÃo os fragmentos foram seccionados com criostato, em sÃrie de 6 micrÃmetros de espessura e submetidos à microscopia de luz. Foram identificados trÃs tipos de mecanorreceptores: Ruffini, Pacini e Golgi, alÃm das terminaÃÃes nervosas livres de acordo com a classificaÃÃo de Freeman e Wike (1967). Em cada grupo foi determinado o nÃmero e a densidade de mecanorreceptores e em seguida foram comparados os resultados. Concluiu-se que o predomÃnio das terminaÃÃes tipo Pacini no gurpo controle foi significante quando comparado com os corpÃsculos tipo Rufini (p < 0,01) e Golgi (p < 0,001).Enquanto que no grupo artrose a densidade do tipo Golgi foi menor que o tipo Pacini(p < 0,001) e terminaÃÃes nervosas livres( p < 0,01).Por outro lado, quando comparados a densidade total das terminaÃÃes nervosas nos dois grupos observou-se uma reduÃÃo significante nos quadris artrÃsicos (p = 0.008). Isto sugere fortemente que a integridade do sistema proprioceptivo parece sofrer modificaÃÃes em quadris artrÃsicos, como conseqÃÃncia da reduÃÃo do nÃmero de terminaÃÃes nervosas. Estudos eletrofisiolÃgicos futuros serÃo necessÃrios para definir o papel das terminaÃÃes nervosas e o padrÃo proprioceptivo do quadril normal do quadril artrÃsico. / Proprioceptive studies have reported growing interest in investigators in the last few decades. This is result of relevant integrity of this system in the treatment of orthopedic diseases. Mechanoreceptors and free nerve endings are the microscopy basic units from proprioception and peripheral nervous system. They transmit to the central nervous system physiological information with detection threshold when the joint are submitted to mechanical stress. The presence and density of mechanoreceptors were investigated in the capsule, teres ligament and acetabulum labrum from 45 hips joint. Of these 30 were obtained from male patients (Group I) with secondary arthrosis at open arthoplasty and 15 of fresh cadaver with normal hip joint (Group II). The mean age of group I was 56,6 and the group II was 36,5 years. The fragments obtained were measured up to 2mm and stained with gold chloride 1%. After fixation the fragments were sectioned with cryostat at serial sections of 6Âm thickness and examined using light microscopy. We identified 3 types of mechanoreceptors: Ruffini, Pacini and Golgi corpuscles, as well free nerve endings according of Freeman and Wike classified 1967. Each group was determined the number and density of mechanoreceptors and then it was compared. We conclude that the number of Pacini type was significative when it was compared with Rufini ( p < 0,01 ) and Golgi types (p< 0,001), in the normal group. However, the denstity of the Golgi type was minor compared to the Pacini ( p < 0,001) and free nerve ending ( p < 0,01 ) in the arthrosis group. Therefore, when the density total of nerve endings in normal hips were compared with arthosis hips we founded that the number decreased (p = 0.008). This is suggests strongly that the integrity of proprioceptive system seems to be modified by arthosis in consequence of nerve endings numbers. In the future, electrophysiological studies will be necessary to determine the rule of nerve endings and proprioceptive system in the normal and arthosis hip.
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Modulation of Neuropeptide Release via Voltage-Dependent and -Independent Signaling in Isolated Neurohypophysial Terminals: a DissertationVelazquez-Marrero, Cristina M. 28 April 2008 (has links)
This thesis details my examination of several mechanisms for modulation of neuropeptide release via voltage-dependent and voltage-independent intraterminal signaling in isolated neurohypophysial terminals. The first part of this work characterizes depolarization-induced neuropeptide release in the absence of extracellular calcium. The goal of this project was to examine the relationship between depolarization-induced release of intracellular calcium stores and depolarization-secretion coupling of neuropeptides. We demonstrate that depolarization in the absence of extracellular calcium induced by either High K+ or electrical stimulation induces a rise in [Ca2+]i and subsequent neuropeptide release from Hypothalamic Neurohypophysial System (HNS) terminals. A portion of extracellular calcium-independent neuropeptide release is due to intraterminal calcium, but the remaining depolarization-induced release may be due to calcium-independent voltage-dependent (CIVD) release (Zhang and Zhou, 2002; Zhang et al., 2004; Yang et al., 2005). Nevertheless, our results clearly show that extracellular calcium is notnecessary for depolarization-induced neuropeptide secretion from these CNS terminals.
In addition, I investigated the role of internal calcium stores in mediating μ-opioid inhibition of voltage-gated calcium channels (VGCCs). Inhibition of VGCCs via μ-opioid agonists has been shown to reduce neuropeptide release in response to High K+ stimulation of isolated terminals (Bicknell et al., 1985b; Russell et al., 1993; van Wimersma Greidanus and van de Heijning, 1993; Munro et al., 1994; Ortiz-Miranda et al., 2003; Russell et al., 2003; Ortiz-Miranda et al., 2005). My findings show μ-opioid inhibition, of VGCC and High K+-mediated rise in [Ca2+]i, are via a voltage-independent diffusible second-messenger targeting release of calcium from ryanodine-sensitive stores, possibly mediated via the cyclic ADP ribose signaling pathway.
Furthermore, I detail a different intracellular messenger pathway mediating the κ-opioid inhibition of VGCC and High K+-mediated rise in [Ca2+]ii. In contrast to the μ-opioid inhibition, κ-receptor activation is coupled to a voltage-dependent membrane-delimited pathway. Inhibition of neuropeptide release via both endogenous and exogenous κ-opioid agonists has been extensively studied (Bicknell et al., 1985a; Nordmann et al., 1986a; Wammack and Racke, 1988; Munro et al., 1994; Ingram et al., 1996; Rusin et al., 1997a). My investigation shows that the κ-inhibition of VGCC is voltage-dependent and is furthermore, relieved within the context of a physiological burst of action potentials (APs). This physiologically-evoked, activity-dependent modulation of VGCC and subsequent release, represents an important mechanism for short-term synaptic plasticity at the level of the terminals. Given the ubiquitous nature of voltage-dependent G-protein signaling in the CNS, our results may prove important in understanding modulatory effects of specific bursting patterns throughout the CNS. In the last 30 years the neurohypophysial system has proven to be an excellent system to study the complexities of depolarization-secretion coupling (DSC). There have been many advances in our understanding of the underlying mechanisms involved and their physiological implications. The current work focuses on two important features of DSC; voltage and calcium. Although in many ways these two are intrinsically linked through VGCC activation, we have found that in isolated HNS terminals that is not always the case. We have further found that when voltage and calcium influx are linked during DSC, modulation by opioids may or may not be linked to activity-dependent relief depending on the opioid receptor activated. This finding has important implications in neuropeptide release during patterned stimulation in vivo. As I will discuss further, many factors play into the complexities of the regulatory mechanisms involving release. As investigations into this remarkable field continue, I hope to have contributed a valuable piece to the puzzle.
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Quantificação digital da imunoexpressão de receptores adrenérgicos e terminações nervosas no detrusor de portadores da síndrome de prune belly / Digital quantification of the imunoexpression of adrenergic receptors and nervous terminations in the detrusor of patients with prune belly syndromeMonteiro, Edison Daniel Schneider 19 March 2008 (has links)
INTRODUÇÃO: A síndrome de prune belly (PBS) é caracterizada por uma tríade com flacidez da parede abdominal, criptorquidia bilateral e malformações do trato urinário que compreende bexiga de capacidade aumentada, com complacência elevada, hipossensibilidade, hipocontratilidade, com divertículo ou fístula uracal e resíduo pós-miccional elevado. Alguns autores recomendam tratamento clínico, porém outros propõe correção cirúrgica, com reconstrução da via urinária incluindo ureteroplastia e cistoplastia redutoras, orquidopexia e abdominoplastia. Mesmo após a cirurgia, alguns doentes necessitam de cateterismo limpo intermitente. A inervação vesical determina seu funcionamento, mediado por neuroreceptores na junção neuromuscular. Os adrenoreceptores a1 estão relacionados à contratilidade detrusora e o b3 ao seu relaxamento, e certas condições como obstrução infravesical levam à hiperexpressão de receptores a1. O objetivo da presente pesquisa é verificar se no detrusor de doentes com PBS há alteração na densidade de terminações nervosas, hiper ou hipoexpressão de receptores adrenérgicos a1a, a1b, a1d e b3 e proporção anormal dos tecidos muscular e conectivo. MÉTODO Trata-se de estudo retrospectivo de caso-controle que envolveu 14 espécimes de detrusor de doentes com PBS operados entre 1985 a 2005 no Hospital das Clínicas da FMUSP. Dois grupos foram constituídos como controle: 13 fragmentos de bexiga de doentes submetidos à prostatectomia radical no Departamento de Urologia da Universidade de Mainz, com urodinâmica pré-operatória normal (GC1), e cinco fragmentos de bexiga de crianças submetidas à necrópsia no SVOC-USP, sem anomalias neurológicas e de trato urinário. A coloração de van Gieson foi realizada para análise da proporção músculo/tecido conectivo, e a reação imunohistoquímica para os anticorpos policlonais anti-proteína S100 e antiadrenoreceptores a1a, a1b, a1d e b3. A coloração castanho foi considerada evidência da expressão do adrenoreceptor na célula. Cinco a dez imagens digitais foram tomadas por meio de câmara digital e microscopia óptica. Estas foram analisadas pelo programa Adobe Photoshop CS2Ò. A quantidade relativa de receptores foi calculada e a análise estatística realizada pelos testes Kruskal-Wallis e Mann-Whitney. RESULTADOS A média de idade foi de 1,28 ± 1,14 ano no grupo caso (PBS), e de 64 ± 5,22 anos e 1,41 ± 1,11 ano, nos grupos GC1 e GC2, respectivamente. A mediana da relação músculo/tecido conectivo foi de 1,08 para o grupo PBS, 1,59 para o GC1 e para o GC2 de 1,28 (p=0,173). A mediana da proporção S100/tecido muscular foi de 0,21 para o grupo caso (PBS), de 0,20 para o GC1 e para o grupo GC2 de 0,01 (p=0,003). A mediana da relação a1a/tecido muscular foi de 0,06 para o grupo PBS, de 0,16 para o GC1 e para o grupo GC2 de 0,14 (p=0,026). Para a1b, as medianas foram 0,06 no grupo PBS, 0,006 no GC1 e 0,007 no GC2 (p=0,781). No a1d, as medianas foram 0,04 (PBS), 0,04 (GC1) e 0,05 (GC2) (p=0,618). Com relação ao b3, as medianas foram 0,07 no grupo PBS, 0,14 no GC1 e 0,10 no GC2 (p=0,378). CONCLUSÕES Comparando-se fragmentos de detrusor de doentes com PBS e bexigas normais não se observou alteração na densidade de terminações nervosas. Observou-se hipoexpressão do adrenoreceptor a1a, e não houve alteração dos adrenoreceptores a1b, a1d e b3. Também não se observou alteração entre a proporção de tecido muscular e conectivo no detrusor destes doentes. Investigações adicionais, com diferentes métodos e incluindo outros receptores, são necessárias antes de se aplicar esses conhecimentos na prática clínica. / INTRODUCTION: Prune belly syndrome (PBS) is charactherized by a triad of abdominal wall flaccidity, bilateral criptorchidism and urinary tract malformation, that includes a large-capacity bladder, with high detrusor compliance, low sensibility and contractility, associated to urachal diverticulum or fistula and elevated post void residual volumes. Some autors recommend clinical treatment, but others propose surgery correction, with urinary tract reconstruction, including reductive ureteroplasty and cystoplasty, orchidopexy and abdominoplasty. Even after surgery, some patients need intermittent catheterism. The detrusor innervation determines its function, mediated by neuroceptors at the neuromuscular junction. The a1 adrenoceptors are related to detrusor contractility and b3 to relaxation, and some conditions, like infravesical obstruction, lead to a1 adrenoceptor up-regulation. The objective of this work is to verify whether, in the detrusor from patients with PBS, there is altered nerve density, up or down-regulation of a1a, a1b, a1d and b3 adrenergic receptors and if there is an abnormal proportion between muscle and connective tissue. MATERIALS AND METHODS A retrospective case-control study was performed involving 14 detrusor specimens from patients with PBS, who underwent surgical treatment between 1985 an 2005 at University of São Paulo, Medical School Hospital. Two groups were taken as control: 13 bladder fragments from patients who underwent radical prostatectomy at Department of Urology of Mainz University, with normal urodynamic study prior to the surgery (GC1) and 5 bladder fragments from children submitted to autopsy at SVOC-USP, with no neurological or urinary tract malformation (GC2). Staining was performed using the van Gieson dye to analyse the proportion between muscle and connective tissue, and immunohistochemical reaction was employed, with polyclonal antibodies against S100 protein, as well as a1a, a1b, a1d and b3 adrenoceptors. Brown colour was considered as evidence of adrenoceptor cell expression. Five to ten digital images were captured on an optic microscope with a digital camera. These images were analysed with Adobe Photoshop CS2Ò software. The relative quantity of receptors was calculated and the statistic analysis was done with the Kruskal-Wallis and Mann-Whitney tests. RESULTS Mean age was 1.28 ± 1.14 year in PBS patients, and 64 ± 5.22 yrs. and 1.41 ± 1.11 yrs. in GC1 and GC2, respectively. The median proportion between muscle and connective tissue was 1.08 in PBS, 1.59 in GC1 and in GC2 of 1.28 (p=0.173). The median proportion of S100/muscle area was 0.21 in PBS, 0.20 in GC1 and in GC2 of 0.01 (p=0.003). The median relative quantity of receptors of a1a was 0.06 in PBS, 0.16 in GC1 and 0.14 in GC2 (p=0.026). In a1b, the median values were 0.06 in PBS group, 0.006 in GC1 and 0.007 in GC2 (p=0.781). In a1d, the median values were 0.04 (PBS), 0.04 (GC1) and 0.05 (GC2) (p=0.618). Regarding b3, the median values were 0.07 in PBS, 0.14 in GC1 and 0.10 in GC2 (p=0.378). CONCLUSION Comparing detrusor fragments from patients with PBS and normal bladders, there was no alteration in the density of nerve endings. We observed downregulation of a1a adrenoceptors, but no alteration in the a1b, a1d and b3 receptors. Furthermore, there was no alteration of the proportion between muscle and connective tissue areas. Further investigations, with different methods and including other receptors, are necessary to transfer this knowledge to clinical use.
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A resposta inflamatória na urticária aguda associada a medicamentos: avaliação imunoistoquímica e imunoeletrônica da unidade microvascular da derme / The inflammatory response in acute drug-induced urticaria: immunohistochemistry and ultrastructure study of dermal microsvascular unitCriado, Paulo Ricardo 06 August 2007 (has links)
INTRODUÇÃO: O conhecimento sobre os tipos celulares envolvidos na patogenia da urticária constitui um elemento essencial para a compreensão da fisiopatologia desta doença. Poucos autores têm dado atenção às interações entre mastócitos e dendrócitos da derme na urticária. Os objetivos deste estudo são orientados no sentido de descreverem-se os tipos de degranulação mastocitária na urticária aguda associada a medicamentos, e o de analisarem-se as interações entre dendrócitos da derme e mastócitos. MÉTODOS: Sete doentes com urticária aguda associada com medicamentos foram incluídos neste estudo. Foram obtidas biopsias cutâneas das lesões urticadas e da pele aparentemente normal destes doentes. Os quatorze fragmentos coletados foram divididos em duas partes (28 fragmentos): uma das partes foi enviada para processamento pela coloração de hematoxilina-eosina, para a coloração de Azul de Toluidina e reações de imunoistoquímica com anticorpos anti-CD34, antifator XIIIA (anti- FXIIIa) e antitriptase e o outro fragmento foi processado para uso na microscopia imunoeletrônica, utilizando-se anticorpos para triptase e FXIIIa, além de dupla imunomarcação com ouro com o uso de anticorpos antitriptase e anti-FXIIIa. RESULTADOS: células imunomarcadas com anticorpos anti-CD34 foram observadas de forma esparsa na derme superficial e de forma mais proeminente na derme reticular. Havia múltiplos dendrócitos dérmicos FXIIIa+ na derme superficial e média, dispersos nas regiões subepidérmicas e em torno doa vasos da derme, tanto na pele urticada com na pele aparentemente normal. O número destas células foi similar nos dois grupos de amostras. Não houve diferença estatística entre o número de células triptase-positivas na pele aparentemente normal e na pele urticada, em todos os doentes. Nós observamos mastócitos íntegros na maioria das amostras da pele aparentemente normal. Tanto as amostras de pele aparentemente normal quanto as amostras de pele urticada apresentavam mastócitos em processo de degranulação do tipo anafilático, com inúmeros grânulos extruídos. Após a dupla imunomarcação com ouro, na imuno-microscopia eletrônica de transmissão foram observadas partículas de ouro de 10 nm (FXIIIa) e 15 nm (Triptase) marcando concomitante os grânulos dos mastócitos indicando que tanto a triptase como o FXIIIa encontraram-se presentes nos grânulos destas células. De forma interessante, nós encontramos uma forte evidência de que grânulos contendo tanto FXIIIa, como triptase, extruídos dos mastócitos são fagocitados pelos dendrócitos da derme. CONCLUSÕES: na urticária aguda associada a medicamentos o padrão de degranulação observado foi do tipo anafilático. Este estudo constitui a primeira demonstração da expressão do FXIIIa nos grânulos intracitoplasmáticos e nos grânulos extruídos dos mastócitos, dispersos na matriz extracelular, nos doentes com urticária aguda associada a medicamentos. Outro fato inédito foi a demonstração da fagocitose dos grânulos extruídos dos mastócitos pelos dendrócitos da derme FXIIIa+ / BACKGROUND: The knowledge about the cell types involved in urticaria is an essential element for understanding the pathophysiology of this disease. Few authors have been attempting on interactions among mast cells and dermal dendrocytes in urticaria. The aims of this study are to describe the types of mast cell degranulation in drug-induced acute, besides to analyze the interactions between mast cell and dermal dendrocyte in urticaria. METHODS: Seven patients with drug-induced acute urticaria were enrolled in the study. We token skin biopsies of urticarial lesion and apparently normal skin. The fourteen fragments collected were divided into two parts (28 sections): one to haematoxylin-eosin stain, Toluidine blue stain and immunohistochemisty reactions with anti-CD34, anti-FXIIIa and anti-tryptase antibodies and other part to immunogold electron microscopy using single antibodies to tryptase and FXIIIa, besides double immunogold labelling with anti-tryptase and anti-FXIIIa. RESULTS: immunolabelled CD34+ cells were observed scattered in the superficial dermis and more prominent in the reticular dermis. There were multiple FXIIIa+ dermal dendrocytes in upper and mid dermis, dispersed in subepidermal areas and around blood vessels, both in apparently normal skin and urticarial lesion of drug-induced acute urticaria. The number of these cells was similar in both groups. There was no difference in tryptase positive cells number between apparently normal skin and urticarial lesions, in all patients. We observed intact mast cells in the majority of the sections of the apparently normal skin. Some sections demonstrated a few mast cells in degranulation process, in anaphylactic degranulation type. In urticarial lesions, several mast cells showed degranulation process, in anaphylactic degranulation type. After double immunogold staining, 10 nm (FXIIIa) and 15 nm (Tryptase) gold particles were seen together over the granules in mast cells indicating that tryptase and FXIIIa are each localized into granules of these cells. Interestingly, we found a strong evidence of than the exocytosed mast cell granules contents both FXIIIa and Tryptase immunolabelled are phagocytised by dermal dendrocytes. CONCLUSIONS: In drug-induced acute urticaria the degranulation pattern of mast cells found was composed by anaphylatic. This is the first report, in acute urtuicaria, concern of the expression of FXIIIa in the cytoplasmic mast cell and in extruded granules into extracellular matrix. Phagocytosis of the extruded mast cell granules by FXIIIa+ dermal dendrocytes in urticaria was observed
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A resposta inflamatória na urticária aguda associada a medicamentos: avaliação imunoistoquímica e imunoeletrônica da unidade microvascular da derme / The inflammatory response in acute drug-induced urticaria: immunohistochemistry and ultrastructure study of dermal microsvascular unitPaulo Ricardo Criado 06 August 2007 (has links)
INTRODUÇÃO: O conhecimento sobre os tipos celulares envolvidos na patogenia da urticária constitui um elemento essencial para a compreensão da fisiopatologia desta doença. Poucos autores têm dado atenção às interações entre mastócitos e dendrócitos da derme na urticária. Os objetivos deste estudo são orientados no sentido de descreverem-se os tipos de degranulação mastocitária na urticária aguda associada a medicamentos, e o de analisarem-se as interações entre dendrócitos da derme e mastócitos. MÉTODOS: Sete doentes com urticária aguda associada com medicamentos foram incluídos neste estudo. Foram obtidas biopsias cutâneas das lesões urticadas e da pele aparentemente normal destes doentes. Os quatorze fragmentos coletados foram divididos em duas partes (28 fragmentos): uma das partes foi enviada para processamento pela coloração de hematoxilina-eosina, para a coloração de Azul de Toluidina e reações de imunoistoquímica com anticorpos anti-CD34, antifator XIIIA (anti- FXIIIa) e antitriptase e o outro fragmento foi processado para uso na microscopia imunoeletrônica, utilizando-se anticorpos para triptase e FXIIIa, além de dupla imunomarcação com ouro com o uso de anticorpos antitriptase e anti-FXIIIa. RESULTADOS: células imunomarcadas com anticorpos anti-CD34 foram observadas de forma esparsa na derme superficial e de forma mais proeminente na derme reticular. Havia múltiplos dendrócitos dérmicos FXIIIa+ na derme superficial e média, dispersos nas regiões subepidérmicas e em torno doa vasos da derme, tanto na pele urticada com na pele aparentemente normal. O número destas células foi similar nos dois grupos de amostras. Não houve diferença estatística entre o número de células triptase-positivas na pele aparentemente normal e na pele urticada, em todos os doentes. Nós observamos mastócitos íntegros na maioria das amostras da pele aparentemente normal. Tanto as amostras de pele aparentemente normal quanto as amostras de pele urticada apresentavam mastócitos em processo de degranulação do tipo anafilático, com inúmeros grânulos extruídos. Após a dupla imunomarcação com ouro, na imuno-microscopia eletrônica de transmissão foram observadas partículas de ouro de 10 nm (FXIIIa) e 15 nm (Triptase) marcando concomitante os grânulos dos mastócitos indicando que tanto a triptase como o FXIIIa encontraram-se presentes nos grânulos destas células. De forma interessante, nós encontramos uma forte evidência de que grânulos contendo tanto FXIIIa, como triptase, extruídos dos mastócitos são fagocitados pelos dendrócitos da derme. CONCLUSÕES: na urticária aguda associada a medicamentos o padrão de degranulação observado foi do tipo anafilático. Este estudo constitui a primeira demonstração da expressão do FXIIIa nos grânulos intracitoplasmáticos e nos grânulos extruídos dos mastócitos, dispersos na matriz extracelular, nos doentes com urticária aguda associada a medicamentos. Outro fato inédito foi a demonstração da fagocitose dos grânulos extruídos dos mastócitos pelos dendrócitos da derme FXIIIa+ / BACKGROUND: The knowledge about the cell types involved in urticaria is an essential element for understanding the pathophysiology of this disease. Few authors have been attempting on interactions among mast cells and dermal dendrocytes in urticaria. The aims of this study are to describe the types of mast cell degranulation in drug-induced acute, besides to analyze the interactions between mast cell and dermal dendrocyte in urticaria. METHODS: Seven patients with drug-induced acute urticaria were enrolled in the study. We token skin biopsies of urticarial lesion and apparently normal skin. The fourteen fragments collected were divided into two parts (28 sections): one to haematoxylin-eosin stain, Toluidine blue stain and immunohistochemisty reactions with anti-CD34, anti-FXIIIa and anti-tryptase antibodies and other part to immunogold electron microscopy using single antibodies to tryptase and FXIIIa, besides double immunogold labelling with anti-tryptase and anti-FXIIIa. RESULTS: immunolabelled CD34+ cells were observed scattered in the superficial dermis and more prominent in the reticular dermis. There were multiple FXIIIa+ dermal dendrocytes in upper and mid dermis, dispersed in subepidermal areas and around blood vessels, both in apparently normal skin and urticarial lesion of drug-induced acute urticaria. The number of these cells was similar in both groups. There was no difference in tryptase positive cells number between apparently normal skin and urticarial lesions, in all patients. We observed intact mast cells in the majority of the sections of the apparently normal skin. Some sections demonstrated a few mast cells in degranulation process, in anaphylactic degranulation type. In urticarial lesions, several mast cells showed degranulation process, in anaphylactic degranulation type. After double immunogold staining, 10 nm (FXIIIa) and 15 nm (Tryptase) gold particles were seen together over the granules in mast cells indicating that tryptase and FXIIIa are each localized into granules of these cells. Interestingly, we found a strong evidence of than the exocytosed mast cell granules contents both FXIIIa and Tryptase immunolabelled are phagocytised by dermal dendrocytes. CONCLUSIONS: In drug-induced acute urticaria the degranulation pattern of mast cells found was composed by anaphylatic. This is the first report, in acute urtuicaria, concern of the expression of FXIIIa in the cytoplasmic mast cell and in extruded granules into extracellular matrix. Phagocytosis of the extruded mast cell granules by FXIIIa+ dermal dendrocytes in urticaria was observed
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Quantificação digital da imunoexpressão de receptores adrenérgicos e terminações nervosas no detrusor de portadores da síndrome de prune belly / Digital quantification of the imunoexpression of adrenergic receptors and nervous terminations in the detrusor of patients with prune belly syndromeEdison Daniel Schneider Monteiro 19 March 2008 (has links)
INTRODUÇÃO: A síndrome de prune belly (PBS) é caracterizada por uma tríade com flacidez da parede abdominal, criptorquidia bilateral e malformações do trato urinário que compreende bexiga de capacidade aumentada, com complacência elevada, hipossensibilidade, hipocontratilidade, com divertículo ou fístula uracal e resíduo pós-miccional elevado. Alguns autores recomendam tratamento clínico, porém outros propõe correção cirúrgica, com reconstrução da via urinária incluindo ureteroplastia e cistoplastia redutoras, orquidopexia e abdominoplastia. Mesmo após a cirurgia, alguns doentes necessitam de cateterismo limpo intermitente. A inervação vesical determina seu funcionamento, mediado por neuroreceptores na junção neuromuscular. Os adrenoreceptores a1 estão relacionados à contratilidade detrusora e o b3 ao seu relaxamento, e certas condições como obstrução infravesical levam à hiperexpressão de receptores a1. O objetivo da presente pesquisa é verificar se no detrusor de doentes com PBS há alteração na densidade de terminações nervosas, hiper ou hipoexpressão de receptores adrenérgicos a1a, a1b, a1d e b3 e proporção anormal dos tecidos muscular e conectivo. MÉTODO Trata-se de estudo retrospectivo de caso-controle que envolveu 14 espécimes de detrusor de doentes com PBS operados entre 1985 a 2005 no Hospital das Clínicas da FMUSP. Dois grupos foram constituídos como controle: 13 fragmentos de bexiga de doentes submetidos à prostatectomia radical no Departamento de Urologia da Universidade de Mainz, com urodinâmica pré-operatória normal (GC1), e cinco fragmentos de bexiga de crianças submetidas à necrópsia no SVOC-USP, sem anomalias neurológicas e de trato urinário. A coloração de van Gieson foi realizada para análise da proporção músculo/tecido conectivo, e a reação imunohistoquímica para os anticorpos policlonais anti-proteína S100 e antiadrenoreceptores a1a, a1b, a1d e b3. A coloração castanho foi considerada evidência da expressão do adrenoreceptor na célula. Cinco a dez imagens digitais foram tomadas por meio de câmara digital e microscopia óptica. Estas foram analisadas pelo programa Adobe Photoshop CS2Ò. A quantidade relativa de receptores foi calculada e a análise estatística realizada pelos testes Kruskal-Wallis e Mann-Whitney. RESULTADOS A média de idade foi de 1,28 ± 1,14 ano no grupo caso (PBS), e de 64 ± 5,22 anos e 1,41 ± 1,11 ano, nos grupos GC1 e GC2, respectivamente. A mediana da relação músculo/tecido conectivo foi de 1,08 para o grupo PBS, 1,59 para o GC1 e para o GC2 de 1,28 (p=0,173). A mediana da proporção S100/tecido muscular foi de 0,21 para o grupo caso (PBS), de 0,20 para o GC1 e para o grupo GC2 de 0,01 (p=0,003). A mediana da relação a1a/tecido muscular foi de 0,06 para o grupo PBS, de 0,16 para o GC1 e para o grupo GC2 de 0,14 (p=0,026). Para a1b, as medianas foram 0,06 no grupo PBS, 0,006 no GC1 e 0,007 no GC2 (p=0,781). No a1d, as medianas foram 0,04 (PBS), 0,04 (GC1) e 0,05 (GC2) (p=0,618). Com relação ao b3, as medianas foram 0,07 no grupo PBS, 0,14 no GC1 e 0,10 no GC2 (p=0,378). CONCLUSÕES Comparando-se fragmentos de detrusor de doentes com PBS e bexigas normais não se observou alteração na densidade de terminações nervosas. Observou-se hipoexpressão do adrenoreceptor a1a, e não houve alteração dos adrenoreceptores a1b, a1d e b3. Também não se observou alteração entre a proporção de tecido muscular e conectivo no detrusor destes doentes. Investigações adicionais, com diferentes métodos e incluindo outros receptores, são necessárias antes de se aplicar esses conhecimentos na prática clínica. / INTRODUCTION: Prune belly syndrome (PBS) is charactherized by a triad of abdominal wall flaccidity, bilateral criptorchidism and urinary tract malformation, that includes a large-capacity bladder, with high detrusor compliance, low sensibility and contractility, associated to urachal diverticulum or fistula and elevated post void residual volumes. Some autors recommend clinical treatment, but others propose surgery correction, with urinary tract reconstruction, including reductive ureteroplasty and cystoplasty, orchidopexy and abdominoplasty. Even after surgery, some patients need intermittent catheterism. The detrusor innervation determines its function, mediated by neuroceptors at the neuromuscular junction. The a1 adrenoceptors are related to detrusor contractility and b3 to relaxation, and some conditions, like infravesical obstruction, lead to a1 adrenoceptor up-regulation. The objective of this work is to verify whether, in the detrusor from patients with PBS, there is altered nerve density, up or down-regulation of a1a, a1b, a1d and b3 adrenergic receptors and if there is an abnormal proportion between muscle and connective tissue. MATERIALS AND METHODS A retrospective case-control study was performed involving 14 detrusor specimens from patients with PBS, who underwent surgical treatment between 1985 an 2005 at University of São Paulo, Medical School Hospital. Two groups were taken as control: 13 bladder fragments from patients who underwent radical prostatectomy at Department of Urology of Mainz University, with normal urodynamic study prior to the surgery (GC1) and 5 bladder fragments from children submitted to autopsy at SVOC-USP, with no neurological or urinary tract malformation (GC2). Staining was performed using the van Gieson dye to analyse the proportion between muscle and connective tissue, and immunohistochemical reaction was employed, with polyclonal antibodies against S100 protein, as well as a1a, a1b, a1d and b3 adrenoceptors. Brown colour was considered as evidence of adrenoceptor cell expression. Five to ten digital images were captured on an optic microscope with a digital camera. These images were analysed with Adobe Photoshop CS2Ò software. The relative quantity of receptors was calculated and the statistic analysis was done with the Kruskal-Wallis and Mann-Whitney tests. RESULTS Mean age was 1.28 ± 1.14 year in PBS patients, and 64 ± 5.22 yrs. and 1.41 ± 1.11 yrs. in GC1 and GC2, respectively. The median proportion between muscle and connective tissue was 1.08 in PBS, 1.59 in GC1 and in GC2 of 1.28 (p=0.173). The median proportion of S100/muscle area was 0.21 in PBS, 0.20 in GC1 and in GC2 of 0.01 (p=0.003). The median relative quantity of receptors of a1a was 0.06 in PBS, 0.16 in GC1 and 0.14 in GC2 (p=0.026). In a1b, the median values were 0.06 in PBS group, 0.006 in GC1 and 0.007 in GC2 (p=0.781). In a1d, the median values were 0.04 (PBS), 0.04 (GC1) and 0.05 (GC2) (p=0.618). Regarding b3, the median values were 0.07 in PBS, 0.14 in GC1 and 0.10 in GC2 (p=0.378). CONCLUSION Comparing detrusor fragments from patients with PBS and normal bladders, there was no alteration in the density of nerve endings. We observed downregulation of a1a adrenoceptors, but no alteration in the a1b, a1d and b3 receptors. Furthermore, there was no alteration of the proportion between muscle and connective tissue areas. Further investigations, with different methods and including other receptors, are necessary to transfer this knowledge to clinical use.
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