Identification in immuno-electron microscopy of solitary multi-polar peripheral neurons of adult Opisthorchis viverrini by antibody against rat diacylglycerol kinase / Identificación por microscopía imuno-electrónica de las neuronas periféricas multipolares solitarias del Opisthorchis viverrini adulto mediante anticuerpo contra la diacilglicerol kinasa de ratas

By utilizing the antibody for rat DGKz a substantial number of immunopositive cells were found in the OV (Opisthorchis viverrini). The immunopositive cells appeared solitarily and they were distributed rather symmetrically to the longitudinal axis of the OV. Some of them were located in close proximity to internal organs such as uterus, ovary, testes, vitelline glands and guts. The immunostained cells extended tapering processes horizontally or obliquely to the OV longitudinal axis. In immuno-electron microscopy, the immunopositive cells were characterized by intensely immunostained mitochondria and weakly immunostained cytoplasm and immunonegative chromatin-poor nucleus. Vacuoles of various sizes without the immunoreactivity were also contained in the cells. Thin cellular processes without the immunoreactivity were found to enclose thinly the entire surfaces of the immunostained cells and processes, and they were in continuity with the interstitial partition-like processes which contained nuclei and aggregation of microfibrils at some distance from the cytoplasmic envelopes. The present finding suggests the possibility that the immunostained cells were peripheral neurons enveloped by peripheral glia and that the glia are of mesenchymal origin because of their cytoplasmic continuity to the interstitial partition-like processes. The motor or sensory nature of the neurons remains to be elucidated.

Mediante el uso del anticuerpos DGK para rata se determinó un número considerable de células inmunopositivas en el Opisthorchis viverrini (OV). Las células inmunopositivas aparecían solitarias y se distribuían simétricamente al eje longitudinal de la OV. Algunas estaban ubicadas en las proximidades de los órganos internos como el útero, ovarios, testículos, glándulas vitelinas e intestino. Las células inmunoteñidas extendían sus procesos horizontalmente u oblicuamente al eje longitudinal de la OV. Por microscopía inmunoelectrónica, las células inmunopositivas se caracterizaron por presentar mitocondrias intensamente teñidas, citoplasma con tinción débil e inmunonegatividad en núcleos pobres en cromatina. También se observó en las células, vacuolas de diversos tamaños sin inmunorreactividad. Se encontraron procesos celulares sin inmunorreactividad para cerrar finamente todas las superficies de las células y procesos, y se continuaron con los procesos de partición intersticiales que contenían núcleos y agregación de microfibrillas a cierta distancia de las envolturas citoplásmicas. El presente hallazgo sugiere la posibilidad de que las células inmunoteñidas son neuronas periféricas envueltas por glia periférica y que la glía presenta origen mesenquimal debido a su continuidad citoplasmática con los procesos de partición intersticiales. La naturaleza motora o sensorial de las neuronas aún no se ha dilucidado.

Immunoelectron microscopy study of superficial skin nerves in drug-induced acute urticaria / Estudo de microscopia imunoeletrônica dos nervos superficiais da pele na urticária aguda induzida por medicamentos

BACKGROUND: Few studies have evaluated the ultrastructure of the superficial skin nerves in urticaria. OBJECTIVE: The objective of this study was to describe findings in superficial skin nerves in cases of drug-induced acute urticaria. METHODS: Seven patients with drug-induced acute urticaria were included in the study. Skin biopsies were obtained from the urticarial lesion and from the apparently normal skin. The 14 fragments collected were processed for immunogold electron microscopy using single stains for antitryptase and anti-FXIIIa antibodies, as well as double immunogold labeling for both. RESULTS: Some sections showed mast cells in the process of degranulation. Following double immunogold staining, 10 nm (FXIIIa) and 15 nm (Tryptase) gold particles were found together throughout the granules in mast cells, indicating that tryptase and FXIIIa are located inside each one of the granules of these cells. Interestingly, we found strong evidence of the presence of tryptase and factor XIIIa in the superficial skin nerves of these patients, both in cases of urticarial lesions (wheals) and in the apparently normal skin. CONCLUSIONS: Tryptase and FXIIIa are present in the superficial nerves of the skin in drug-induced acute urticaria. This is the first report of tryptase and FXIIIa expression in the superficial skin nerves of patients with urticaria. Tryptase may be participating in neural activation in these patients, while FXIIIa may be present in the nerves to guarantee the functional integrity of structures.

FUNDAMENTOS: Poucos autores têm estudado a ultraestrutura dos nervos superficiais na urticária. OBJETIVO: Descrever os achados nos nervos cutâneos superficiais em casos de urticária aguda induzida por medicamentos. MÉTODOS: Sete pacientes com urticária aguda induzida por medicamentos foram incluídos no estudo. Foram obtidas biopsias da pele da lesão urticariforme e da pele aparentemente normal. Os 14 fragmentos coletados foram processados usando imunomarcação com ouro para anticorpos anti-triptase e anti-FXIIIa separadamente, além da dupla imunomarcação com ambos anticorpos. A seguir as amostras foram submetidas à análise por microscopia imunoeletrônica. RESULTADOS: Alguns cortes demonstraram mastócitos em processo de degranulação. Após a imunomarcação dupla, partículas de ouro de 10 nm (FXIIIa) e partículas de ouro de 15 nm (Triptase) apresentavam-se juntas em grânulos de mastócitos indicando que a triptase e o FXIIIa se localizam dentro de cada um dos grânulos dessas células. Curiosamente, foi encontrada uma forte evidência da presença da triptase e do fator XIIIa nos nervos superficiais dos pacientes avaliados, tanto em lesões urticadas, como na pele aparentemente normal. CONCLUSÕES: A triptase e o FXIIIa estão presentes nos nervos superficiais da pele na urticária aguda medicamentosa. Este é o primeiro relato da expressão de triptase e de FXIIIa nos nervos superficiais na urticária. A triptase poderia estar participando da ativação neural nos pacientes estudados. O FXIIIa poderia estar presente nos nervos, com a finalidade de manter a integridade funcional dessas estruturas.

Viability of nerve grafts preseved in different storage medium

We compared the structural features of nerve segments stored in two different solutions previous and after autologous transplantation. Male Wistar rats were divided into groups to obtain normal tibial nerves, freshly transplanted nerves, and nerves stored in Wisconsin/Belzer or Collins solution for 24 or 72 h at 4ºC and transplanted. Stored and transplanted segments were processed for morphologic and morphometric analysis. The cross-sections of segments stored in Wisconsin/Belzer and Collins solution presented aspects similar to that of normal nerves. The density of large-caliber myelinated axons was higher in grafts stored in Wisconsin/Belzer solution than in those preserved in Collins solution. But the density of myelinated axons regenerated through these grafts was around 80% to that registered in the fresh and Wisconsin/Belzer preserved grafts. Moreover, no significant differences in the morphometric parameters were observed between groups. Our data confirm the efficacy of Wisconsin/Belzer to nerve graft preservation and stimulate more detailed physiological, biochemical and molecular studies to rationalize the employment of less expensive and handful storage solutions for short term preservation of peripheral nerve grafts.

Clinicopathologic analysis of 124 biopsy-proven peripheral nerve diseases

We reviewed dinical, histological and ultrastructural findings of 124 cases of sural nerve biopsy specimens to delineate the trends of peripheral nerve diseases in our institute. Eighty-one were men and 43 were women. We categorized them into five groups: specific diagnosis (66 cases, 53.2%), axonal degeneration type (47 cases, 37.9%), demyelinating type (4 cases, 3.2%), mixed axonal degeneration-demyelinating type (6 cases, 4.8%) and normal (1 case, 0.9%). Cases with specific diagnosis included 21 inflammatory demyelinating polyneuropathy (15 chronic inflammatory demyelinating polyradiculoneuropathy, 6 Guillain-Barre disease), 13 hereditary motor and sensory neuropathy (7 Charcot-Marie-Tooth type I, 6 Charcot-Marie-Tooth type II), 10 vasculitis, 6 toxic neuropathy, 4 leprosy, 3 diabetic neuropathy, 2 alcoholic neuropathy, 1 Fabry's disease and other specific diseases (5 cases). In our cases, the proportion of specific diagnoses was higher, while the proportion of demyelinating peripheral neuropathies and normal were lower than those of Western series. The results of this study indicate that 1) a dose clinicopathologic correlation is important to make a precise diagnosis of peripheral nerve biopsy, 2) Biopsy under strict indication may reduce unnecessary histologic examination, 3) There is no difference in disease pattern of peripheral neuropathy between Western people and Koreans.

Electrophysiological and ultra structural study of nerve biopsies from cytotoxic induced peripheral neuropthy in children

Forty male children with non-Hodgkin's lymphoma [NHL] were included in this study. Their mean age was 9.83 years +/- 1.59 and disease duration was 1.68 years +/- 0.71. They were on vincristine [VCR] that was given in courses, each course for 2-4 weeks during which a single intravenous dose of VCR was given [1[1/2] mg/m[2]] for 8 courses. We had two groups of patients: The first group included 20 children who had ended their courses [8 courses]. The second group comprised 20 children who were still on treatment seven children had four courses, six children had three courses, and seven children had two courses. A control group of 10 healthy volunteers who were matched for age and sex was added. Six patients of both groups had infrequent muscle cramps. All patients of both group were neurologically free. Fifteen children of the first group and only two of the second group had evidence of denervation potentials on electromyographic [EMG] study. Also motor nerve conduction velocity [MNCV] and sensory nerve conduction velocity [SNCV] showed more delay in the first group than in the second group. The mean +/- SD of MNCV was 33.2 m. /sec. +/- 2.8 and 38.1 +/- 4.2 in both groups respectively. SNCV was 29.9 m. /sec +/- 3.1 and 43.2 +/- 5.6 in both groups respectively. Ultrastructural study of nerve biopsies showed more degeneration of the nerve axons with fibrosis as well as degenerative changes in some areas through the myelin sheath in the first group than that of the second group as compared to the normal findings in the control group

Ultrastructural study of the dermal nerves in the cutaneous macular lesions of patients with early leprosy

Thirteen biopsies of macular lesions of early leprosy patients were studied ultrastructurally with transmission electron microscopy (TEM). All of the biopsies displayed at least one dermal nerve partially or completely encircled by mononuclear cells in the conventional histopathological study with light microscopy. The patients'diagnosis varied from indeterminate leprosy to borderline tuberculoide (BT). In the ultrastructural study, twenty-seven dermal nerve branches were found in the thirteen biopsies. Twenty dermal nerve branches in eleven biopsies were found to display no inflammatory involvement. Seven nerves in seven biopsies were morphologically associated with mononuclear leukocytic cells. Four biopsies exhibited nerves with and without inflammatory involvement concomitantly. Three nerves showed morphological evidence of endoneurial fibrosis, not morphologically associated with the inflammatory process at least in the sections examined. No detectable axional and Schwann cell ultra strutural changes even in the twenty-seven nerves were found. The sensorial loss exhibited by the patients before the institution of treatment was completely reversed in eight patients after the end of multidrug therapy regimen. These findings suggest that sensory loss in the early stages of leprosy may be caused by reversible pathological mechanisms, rather than anatomical damage. It is also possible, concerning the mechanisms of nerve damage in leprosy, to speculate on the existence of a pathological process which may precede the inflammation.

Importância da inervaçåo macro e microscópia do joelho no tratamento das gonalgias pela acupuntura e sua relaçåo com a teoria dos canais de energia e pontos de acupuntura da medicina chinesa / The importance of knee inervation (macro and micro) during acupuntural treatment and it's relationship with energy channels theory and the chosen acunputural points in Chinese medicine

O conhecimento da inervaçåo macro e micróscopia do joelhoé importante para compreender-se a origem e o mecanismo das gonalgias. Såo descritos os principais ramos dos nervos articulares e os nociceptores, que eståo presentes nas diferentes estruturas anatomicas que compöem o joelho. Ambos såo importantes na gênese dos arcos reflexos sômato-somático e sômato-viscerais, estes reflexos explicam as gonalgias bcom alteraçöes estruturais do membro inferior e com as manifestaçöes viscerais e psíquicas. Os autores relacionam estes conhecimentos neurofisiológicos com a teoria dos canais de energia e dos pontos de acupuntura da medicina tradicional chinesa, evidenciando-se a importância da inervaçåo e dos nociceptores no mecanismo de açåo das agulhas de acupuntura

Increased incidence of intranuclear rods and unusual cytoplasmic inclusions in clarke neurons in rats subjected to neonatal peripheral nerve injury: an ectron microscopic study

Clarke's nucleus of young adult rats whose right sciatic nerves were crushed in the first postnatal day was examined with the electron microscope and the ultrastructure of Clarke neurons was described on both the control [left] and experimental [right] ides. The general fine tructure of Clarke cells appeared quite similar to that of other neurons. Unlike other neurons. Clarke cells showed an eccentric nucleus and Nissl substance characterized by parallel cisternae of rough endoplasmic reticulum. Three types of intranuclear rods and two distinctive cytoplasmic inclusions, termed fibrillary inclusion bodies and nematosomes, have been identified in Clarke neurons on the experimental side. The nuclear rods consisted of microfilaments or microtubules and were called tubular, filamentous and filamentous.crystalloid rods. The fibrillary inclusion bodies appeared as balls of densely packed neurofilaments enveloped by a multiloculated membrane. The nematosomes consisted of a reticular network of filamentous strands, lacked a limiting membrane and were always present in ribosome-rich areas. On the control side, the cytoplasmic inclusion bodies were not seen, while the intranuclear rods were observed only in two clarke cells. Mother distinctive feature of Clarke cells on the experimental side was the presence of peculiar dendritie profiles packed with mitochondria and glyeogen particles

Neuropatias periféricas: aspectos morfológicos e importância da correlaçäo clínico-patológica / Peripheral neuropathies: morphological aspects and importance of clinicopathological correlation

As neuropatias periféricas säo doenças comumente encontradas, que algumas vêzes se tornam problemas diagnósticos. Embora possam ser desordens isoladas, elas säo mais frequentemente uma manifestaçäo de uma doença sistêmica subjacente. A abordagem diagnóstica das neuropatias é multidisciplinar. Dados clínicos, familiares, laboratoriais e eletrofisiológicos devem ser obtidos antes de ser indicada uma biópsia de nervo, a qual só deve ser realizada após consideraçäo cuidadosa de sua real contribuiçäo para o manejo do paciente. É preciso ser levado em conta que: o procedimento por si só, irá resultar em pelo menos algum grau de déficit neurológico; há situaçöes que, quando típicas, näo justificam a biópsia; os nervos exibem um espectro muito limitado de alteraçöes morfológicas, que säo semelhantes seja qual for a etiologia da neuropatia; e ainda, a biópsia só deve ser realizada se houver condiçöes técnicas apropriadas para o estudo completo do nervo, pois métodos histológicos convencionais säo, num grande número de casos, insuficientes para uma análise adequada. Nesta revisäo, säo apresentados os aspectos normais e patológicos do nervo periférico, as técnicas utilizadas no estudo do nervo, a classificaçäo morfológica das neuropatias e o valor das biópsias no diagnóstico destas afecçöes

Ultrastructural studies of peripheral nerves in lepromatous leprosy patients.

Ultrathin sections of the peripheral nerves taken from three lepromatous leprosy patients (One untreated, other treated and third in ENL reaction) was examined in the electronmicroscope. In the untreated patient, solid M. leprae organism inside the schwann cell and the degeneration of schwann cell was seen. In contrast, the treated patient showed the degeneration of bacilli and myelinated fibres. However, the characteristics of cells in the ENL reaction showed close similarities with the untreated case.