RESUMEN
BACKGROUND: The diagnosis of Hansen disease (HD) can be difficult when acid-fast bacilli are not detected in the patient's skin sample. OBJECTIVE: To demonstrate that detailed morphological analysis of nonspecific inflammatory and/or noninflammatory alterations in dermal nerves as well as skin adnexa in leprosy-suspected biopsy samples could improve the efficacy of histopathological diagnosis. METHODS: Patients with one to five skin lesions were enrolled in the study and classified into three groups by skin histopathology findings: Hansen disease (HD, n = 13), other diseases (OD, n = 11), and inconclusive cases (INC, n = 11). We quantified dermal nerve damage via the nerve lesion index (NLI) and PGP9.5-immunoreactive axon quantitative index in dermal nerves (AQI). We also measured inflammatory involvement of adnexa in cutaneous samples as indirect evidence of HD. RESULTS: We observed a higher median endoneurial inflammatory infiltrate NLI (HD = 0.5; INC = 0; OD = 0; p < 0.001) and more frequent inflammatory involvement of skin adnexa in samples of the HD group compared with those of the INC and OD groups (HD = 7; INC = 1; OD = 0). However, samples from the INC and OD groups also showed inflammatory and noninflammatory damage of dermal nerves, with 2 or more kinds of alterations in nerves in the same sample (respectively: INC = in 1 and 2 samples; OD = in 3 and 5 respectively). The quantification of PGP9.5-immunoreactive axons in dermal nerves revealed no difference between the groups. CONCLUSION: A detailed morphological analysis of cutaneous nerves in lesions with a suspicion of HD enabled us to select patients with nonspecific inflammatory or non-inflammatory lesions in the dermal nerves in the INC and OD groups, so they may be clinically monitored aiming at a possible future diagnosis of the disease. These INC and OD patients cannot have the HD diagnosis definitely excluded, and HD may coexist with another disease as a comorbidity.
ANTECEDENTES: A hanseníase pode ter o seu diagnóstico histopatológico dificultado quando bacilos álcool-ácido resistentes não são encontrados nas amostras de pele dos pacientes. OBJETIVO: Demonstrar que uma análise morfológica detalhada de alterações histopatológicas dos nervos dérmicos pode aumentar a eficácia diagnóstica. MéTODOS: Foram selecionadas amostras de pele de pacientes com uma a cinco lesões suspeitas de hanseníase. Os casos selecionados foram classificados conforme achados histopatológicos: hanseníase (HD, n = 13), casos inconclusivos (INC, n = 11), e outras doenças (OD, n = 11). Quantificamos as lesões dos nervos cutâneos por meio do índice de lesão de nervos (nerve lesion index, NLI, em inglês) e do índice quantitativo de axônios (axon quantitative index, AQI, em inglês) imunorreativos a PGP9.5 nos nervos cutâneos. Também medimos o envolvimento inflamatório dos anexos em amostras de pele como evidência indireta de hanseníase. RESULTADOS: Foram observadas no grupo HD medianas mais altas do NLI com relação a infiltrados inflamatórios endoneurais (HD = 0,5; INC = 0; OD = 0; p < 0,001) e mais alta frequência de acometimento inflamatório de anexos cutâneos (HD = 7; INC = 1; OD = 0). Entretanto, as amostras dos grupos INC e OD também mostraram comprometimento inflamatório e não inflamatório dos nervos cutâneos, com 2 ou mais tipos de alterações de nervos na mesma amostra (respectivamente: INC = 1 e 2; OD = 3 e 5). Não houve diferença significativa na quantidade de axônios endoneurais imunorreativos a PGP9.5 entre os grupos. CONCLUSãO: A análise morfológica detalhada dos nervos cutâneos em lesões suspeitas de hanseníase permitiu selecionar pacientes com lesões inespecíficas inflamatórias ou não inflamatórias nos nervos dérmicos nos grupos INC e OD, para que sejam monitorados clinicamente visando um possível diagnóstico futuro da doença. Esses pacientes INC e OD não podem ter o diagnóstico de HD definitivamente excluído, e a hanseníase pode coexistir com outra doença como uma comorbidade.
Asunto(s)
Inmunohistoquímica , Lepra , Piel , Humanos , Masculino , Femenino , Lepra/patología , Lepra/complicaciones , Persona de Mediana Edad , Adulto , Piel/inervación , Piel/patología , Biopsia , Anciano , Adulto Joven , Ubiquitina Tiolesterasa/análisis , Adolescente , Estadísticas no ParamétricasRESUMEN
BACKGROUND: Spitzoid tumors are a heterogeneous group of melanocytic neoplasms that frequently imposes diagnostic difficulties. Lately, several advances in molecular biology afforded significant discoveries on the pathogenesis of these tumors. BAP1 (BRCA-1 associated protein-1) inactivation and anomalous expression of kinase translocation-related proteins are among the main criteria launched by new classification proposals. Our aim was to systematically assess the immunoexpression of BAP1, ROS1 (receptor tyrosine kinase c-Ros oncogene 1), and ALK (anaplastic lymphoma receptor tyrosine kinase) proteins in an unpublished series of spitzoid tumors. METHODS: Retrospective study based on 47 formalin-fixed paraffin-embedded tissue samples from 3 different institutions. BAP1, ROS1, and ALK immunostains were performed in all cases. We included 27 Spitz tumors without significant abnormality, 15 atypical spitzoid tumors, and 5 spitzoid melanomas. RESULTS: We observed loss of BAP1 nuclear immunolabeling in 4.3% of evaluable cases (2/46), both of them atypical spitzoid tumors. The proportional frequency of BAP1-inactivated cases among atypical spitzoid tumors was 14.2% (2/14). No immunoexpression of ROS1 or ALK was found. CONCLUSIONS: Our study revealed 2 additional BAP1-inactived cases and described its respective frequency. The absence of anomalous expression of translocation-related proteins ALK and ROS1 in this series, composed predominantly of low-grade/low-risk tumors, indicates that translocated spitzoid lesions may not be as prevalent as initially suggested, at least in some populations. Furthermore, our findings encourage additional investigation on unequal occurrence of such immunomarkers among different diagnostic categories of spitzoid neoplasms.
Asunto(s)
Nevo de Células Epitelioides y Fusiformes/diagnóstico , Proteínas Tirosina Quinasas/análisis , Proteínas Proto-Oncogénicas/análisis , Proteínas Tirosina Quinasas Receptoras/análisis , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/patología , Proteínas Supresoras de Tumor/análisis , Ubiquitina Tiolesterasa/análisis , Adolescente , Adulto , Quinasa de Linfoma Anaplásico , Biomarcadores de Tumor/análisis , Niño , Preescolar , Femenino , Humanos , Inmunohistoquímica , Lactante , Masculino , Persona de Mediana Edad , Nevo de Células Epitelioides y Fusiformes/patología , Estudios Retrospectivos , Adulto JovenRESUMEN
The sensory nerve endings of the rat tongue, cheek and palate were studied using immunohistochemical staining and transmission electron microscopy analysis. The specimens were fixed in modified Karnovsky solution and embedded in Spurr resin. Substance P, calcitonin gene-related peptide (CGRP)- and protein gene product 9.5 (PGP b9.5)-containing nerve fibers in the rat tongue, cheek and palate were examined by electronic microscopical analysis and immunohistochemical localization. These fibers run very close to the basal lamina of the epithelium and extend into the filliform and fungiform papillae. Numerous plexiform fibers immunoreactive for substance P, CGRP and PGP 9.5 were found in the connective tissue of mucosa. Electron microscopic observations showed clearly immunostained nerve fibers, which are located very close to the basal lamina of epithelial cells. Some electron-dense granules may be observed in the axoplasms of both substance P and CGRP immunoreactive fibers. Several lamellar corpuscles into the subepithelial connective tissue papillae, Merkel corpuscles and numerous thin unmyelinated and myelinated axons were observed. The terminal axons revealed numerous mitochondria, neurofilaments, microtubules and clear vesicles in the base of axoplasmic protrusions. The lamellar cells showed caveolae and interlamelar spaces filled by amorphous substance. Between the lamellar cells and axoplasmic membrane, and in the adjacent lamellae region, desmosome-type junctions were observed. The quantitative and morphometric analysis showed nerve endings with an average area of 4.83 ± 3.4 µm(2) and 19.4 internal mitochondria in this site and the organized corpuscles with an average area of 79.24 ± 27.24 µm(2) and 24.23 internal mitochondria in this place. All the structures observed are involved in the transmission of pain and mechanoreceptors stimulus of these oral mucosae.
Asunto(s)
Mucosa Bucal/inervación , Terminaciones Nerviosas/ultraestructura , Células Receptoras Sensoriales/ultraestructura , Animales , Axones , Péptido Relacionado con Gen de Calcitonina/análisis , Mejilla/inervación , Técnicas para Inmunoenzimas , Células de Merkel , Microscopía Electrónica de Transmisión , Mitocondrias/ultraestructura , Mucosa Bucal/ultraestructura , Hueso Paladar/inervación , Ratas , Ratas Wistar , Sustancia P/análisis , Lengua/inervación , Ubiquitina Tiolesterasa/análisisRESUMEN
Several reports have shown that cutaneous leishmaniasis lesions are painless, suggesting that Leishmania infection interferes with pain perception. Comparisons of inflammation-induced hyperalgesia between BALB/c and C57BL/6 mice have been little explored in the literature, and comparative data regarding nociception in leishmaniasis are non-existent. In susceptible BALB/c mice and resistant C57BL/6 mice that were intradermally inoculated with a low dose of Leishmania major in the ear, we investigated the variation in nociception over a 12-wk period post-infection and this variation's association with the structure of nerve fibres and the presence of endogenous cytokines that are classically considered hyper- or hypo-nociceptive. Infected BALB/c mice presented susceptibility and severe lesions. Infected C57BL/6 mice exhibited resistance and healing lesions. The immune response involved pro- and anti-inflammatory cytokine secretion, respectively. The infection-induced hypoalgesia in BALB/c mice after wks 9 was accompanied by decreased levels of IL-6 and IL-10 in ear tissue with intact nerves. C57BL/6 mice showed short-lived hyperalgesia in wks 2, which was related to increased local levels of IL-6, KC/CXCL-1, TNF-α and IL-10 and a decrease in nerve density. The increase in pro-inflammatory cytokine IL-6, KC/CXCL-1 and TNF-α levels during hyperalgesia suggested a role for these mediators in afferent nerve sensitisation, which was secondary to the inflammatory damage of nerve fibres stained by PGP 9.5. In contrast, the mechanisms of hypoalgesia may include the downregulation of cytokines, the preservation of the structure of nerve endings, and as yet uninvestigated unidentified differences in neurotransmitter release or a direct role of the parasites in the context of the progressive and permissive inflammatory response of BALB/c mice.
Asunto(s)
Citocinas/análisis , Oído Externo/parasitología , Leishmania major/inmunología , Leishmaniasis Cutánea/fisiopatología , Nocicepción/fisiología , Animales , Citocinas/metabolismo , Regulación hacia Abajo , Oído Externo/inmunología , Oído Externo/inervación , Femenino , Inmunohistoquímica , Leishmania major/patogenicidad , Leishmaniasis Cutánea/complicaciones , Leishmaniasis Cutánea/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Dolor Nociceptivo/etiología , Dolor Nociceptivo/inmunología , Dolor Nociceptivo/fisiopatología , Umbral del Dolor/fisiología , Ubiquitina Tiolesterasa/análisisRESUMEN
Notalgia paraesthetica is a neuropathic pruritus on the back. The aim of this retrospective study was to examine patient characteristics in a consecutive cohort from Brazil and Germany. A total of 65 patients (49 women, 16 men; age range 25-80 years, mean 56.2 ± 12.7 years; median 57.0 years) were investigated in order to determine the spinal or peripheral origin of notalgia paraesthetica. Protein gene product 9.5-positive intraepidermal nerve fibers were significantly reduced in the pruritic compared with the non-lesional area (p < 0.05). In 32.3% of patients, radiological examinations showed a stenosis and in 47.7% a degeneration. A correlation between the radiological findings and the exact dermatomal localization of notalgia paraesthetica was found in 15.7% of the involved areas. The significant reduction in intraepidermal nerve fiber density suggests that damage to the peripheral nerves is a more important aetiological factor than spinal changes in notalgia paraesthetica.
Asunto(s)
Nervios Periféricos/patología , Enfermedades del Sistema Nervioso Periférico/patología , Prurito/patología , Piel/inervación , Adulto , Anciano , Anciano de 80 o más Años , Dorso , Biomarcadores/análisis , Biopsia , Brasil/epidemiología , Femenino , Alemania/epidemiología , Humanos , Inmunohistoquímica , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Nervios Periféricos/química , Enfermedades del Sistema Nervioso Periférico/epidemiología , Prurito/diagnóstico por imagen , Prurito/epidemiología , Estudios Retrospectivos , Piel/patología , Compresión de la Médula Espinal/diagnóstico por imagen , Compresión de la Médula Espinal/epidemiología , Compresión de la Médula Espinal/patología , Nervios Espinales/química , Nervios Espinales/patología , Tomografía Computarizada por Rayos X , Ubiquitina Tiolesterasa/análisisRESUMEN
The proteinase-activated receptor 2 (PAR(2)) is a putative therapeutic target for arthritis. We hypothesized that the early pro-inflammatory effects secondary to its activation in the temporomandibular joint (TMJ) are mediated by neurogenic mechanisms. Immunofluorescence analysis revealed a high degree of neurons expressing PAR(2) in retrogradely labeled trigeminal ganglion neurons. Furthermore, PAR(2) immunoreactivity was observed in the lining layer of the TMJ, co-localizing with the neuronal marker PGP9.5 and substance-P-containing peripheral sensory nerve fibers. The intra-articular injection of PAR(2) agonists into the TMJ triggered a dose-dependent increase in plasma extravasation, neutrophil influx, and induction of mechanical allodynia. The pharmacological blockade of natural killer 1 (NK(1)) receptors abolished PAR(2)-induced plasma extravasation and inhibited neutrophil influx and mechanical allodynia. We conclude that PAR(2) activation is pro-inflammatory in the TMJ, through a neurogenic mechanism involving NK(1) receptors. This suggests that PAR(2) is an important component of innate neuro-immune response in the rat TMJ.
Asunto(s)
Artritis/patología , Receptor PAR-2/análisis , Trastornos de la Articulación Temporomandibular/patología , Animales , Artropatía Neurógena/patología , Inmunidad Innata/inmunología , Inyecciones Intraarticulares , Masculino , Fibras Nerviosas/patología , Neuroinmunomodulación/inmunología , Antagonistas del Receptor de Neuroquinina-1 , Neuronas/patología , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/patología , Oligopéptidos/administración & dosificación , Oligopéptidos/farmacología , Dimensión del Dolor , Piperidinas/farmacología , Plasma , Quinuclidinas/farmacología , Ratas , Ratas Wistar , Receptor PAR-2/agonistas , Células Receptoras Sensoriales/patología , Sustancia P/análisis , Articulación Temporomandibular/inervación , Ganglio del Trigémino/patología , Tripsina/administración & dosificación , Tripsina/farmacología , Ubiquitina Tiolesterasa/análisisRESUMEN
BACKGROUND: Abrikossoff or granular cell tumour (GCT) is a relatively rare neoplasia, benign in most of the cases. It may occur in any part of the human body, but it has an oral location in 70% of the cases. Its origin has been discussed for decades, and it is not yet definitively determined. Immunohistochemical techniques suggest its origin in the Schwann cells, while more recent studies with new markers indicate an origin related to neuroendocrine cells. OBJECTIVE: Contribute to the clarification of histogenesis of oral Abrikossoff tumour studying immunohistochemical marking of 11 oral Brazilian cases. MATERIALS AND METHODS: Samples of tissues from the oral mucosa, tongue and lips placed in paraffin blocks, from eleven patients with a histopathological diagnosis of benign GCT were studied. Four different anti-serums (S-100, vimentin, PGP9.5 and ENE) were used for immunoperoxydase technique. RESULTS: A clear positivity for S-100 protein and vimentin was observed, with markers indicating origin from the Schwann cells. Less intense positivity was found in some cases, for ENE and PGP9.5, which suggests a neuroendocrine origin. CONCLUSIONS: The results obtained suggest an origin from Schwann cells, but also arise the possibility of neuroendocrine origin. New methods and more specific immunohistochemical markers are needed to elucidate the origin of the Abrikossoff tumour.
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Tumor de Células Granulares/patología , Neoplasias de la Boca/patología , Adolescente , Adulto , Biomarcadores de Tumor/análisis , Núcleo Celular/patología , Citoplasma/patología , Femenino , Humanos , Hiperplasia , Inmunohistoquímica , Neoplasias de los Labios/patología , Masculino , Persona de Mediana Edad , Mucosa Bucal/patología , Células Neuroendocrinas/patología , Fosfopiruvato Hidratasa/análisis , Proteínas S100/análisis , Células de Schwann/patología , Neoplasias de la Lengua/patología , Ubiquitina Tiolesterasa/análisis , Vimentina/análisis , Adulto JovenAsunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Tumor de Células Granulares/patología , Anciano , Antígenos CD/análisis , Antígenos de Diferenciación Mielomonocítica/análisis , Neoplasias de la Mama/química , Antígenos CD57/análisis , Calbindina 2 , Femenino , Tumor de Células Granulares/química , Humanos , Inmunohistoquímica , Inhibinas/análisis , Antígeno Ki-67/análisis , Mamografía/métodos , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Proteína G de Unión al Calcio S100/análisis , Proteínas S100/análisis , Ubiquitina Tiolesterasa/análisisRESUMEN
Skin biopsy has become an attractive technique to evaluate the terminal regions of small nerve fibers. There is extensive innervation of the skin by both sensory and autonomic fibers as demonstrated by staining for the pan-axonal marker PGP 9,5. The normal pattern is fundamental before any study, since three different techniques described in the literature with different results. Skin biopsy specimens of 3-mm in diameter were obtained from the distal leg of 30 healthy controls. Median intraepidermal nerve fiber density was 5.3/mm. Skin biopsy may be a useful tool for assessing the topographic extent and degree of nerve fiber damage in sensory neuropathies and may be particularly useful in experimental treatment trials for peripheral neuropathies since, in contrast to standard nerve biopsy, the test can be repeated.
Asunto(s)
Epidermis/inervación , Fibras Nerviosas , Ubiquitina Tiolesterasa/análisis , Adulto , Biopsia , Epidermis/química , Epidermis/ultraestructura , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Fibras Nerviosas/ultraestructuraRESUMEN
O recente método de avaliação das fibras nervosas intraepidérmicas com o PGP 9,5 vem se mostrando de grande utilidade no diagnóstico das neuropatias sensitivas de fibras finas, autonômicas e neuropatias periféricas subclínicas. Devido à variação da técnica relatada na literatura é de fundamental importância uma padronização normal. Estudamos 15 homens e 15 mulheres com média de idade de 34,5 anos. Em todos os voluntários foi realizada biopsia de pele na porção distal da perna. A média da densidade linear das fibras nervosas intraepidérmicas foi 5,3/mm com mediana de 6,0 e desvio padrão de 1,94. Essa técnica possui um grande número de vantagens em relação à biopsia de nervo convencional, é simples, pouco invasiva, reproduzível e pode ser repetida no mesmo paciente para avaliar progressão da neuropatia e possíveis respostas terapêuticas.