Experimental vaccinia virus meningoencephalitis in adult albino mice: Virological,light microscopic and ultrastructural studies

Summary Adult NMRI mice were inoculated intracerebrally with 1.5–5×10² pfu of the neurovirulent strain Ma1 of vaccinia virus. The animals usually became diseased on day 3 and died on day 5 or 6 p.i. Infectivity assay studies on the mouse brains yielded a steeply rising viral titer comparable with that exhibited by several other so-called neurovaccinia strains after intracerebral inoculation. Histological studies revealed a patchy leptomeningitis most apparent over the basal brain surface. Mononuclear elements of varied size were the most common infiltrate cells in the meningitic lesions. These often showed a pronounced hemorrhagic and necrotizing character. In some places, the cellular infiltrates within the subarachnoid space extended along the perforating vessels into the superficial cerebral cortex. By electron microscopy, all essential stages of virus morphogenesis including the discharge of mature virions could be seen in numerous mononuclear phagocytes and adventitial cells. In contrast to this, no unequivocal neuroectodermal cell showed morphologic evidence of productive viral infection. Likewise, no endothelia or vascular myocytes were encountered which contained newly formed virus particles. However, alterations of the walls of blood vessels and signs of disturbed vascular permeability were a frequent finding in the meningeal foci of inflammation as well as in and around the track left by the inoculating needle. Another ultrastructural feature of the meningitic lesions was the widespread occurrence of degenerating or disintegrating infiltrate cells both infected and uninfected ones.The virologic and morphologic findings observed in this model of experimental vaccinia virus meningo-encephalitis are discussed with regard to data previously published by other authors. The propensity of strain Ma1 of vaccinia virus (and most probably of other so-called neurovaccinia strains) for replication in mesenchymal cells in particularly emphasized.     

Cell populations in the lesion of human cutaneous leishmaniasis: A light microscopical,immunohistochemical and ultrastructural study

Summary To characterize the in situ cellular immune response in localized cutaneous leishmaniasis (LCL), the authors studied frozen skin biopsies from 50 patients with LCL due toLeishmania braziliensis guyanensis. A panel of 31 monoclonal antibodies was used, which defined the number and distribution of inflammatory cell subsets. Skin inflammatory infiltrates were composed of T cells (with a local CD4/CD8 ratio of 1.05±0.7 vs 1.48±0.3 in peripheral blood), macrophages and a smaller number of B cells, natural killer cells and granulocytes. Most of the T cells expressed activation markers (interleukin-2 and transferrin receptors, HLA-DR⁺) and an increase in T-cell-receptor expression was noted. Analysis of the CD4⁺ subpopulations with newly available reagents showed that helper T cells (CD4⁺CD45RO⁺) exceeded the suppressor/inducer subset (CD4⁺CD45RA⁺) by 1.41. There were no differences between local immune variables from patients with primary infection (45 patients) and those with recurrence (5). In 7 patients, biopsies were analysed before and 1 month after specific treatment, and did not show significant differences except for a small increase of dermal CD1a⁺ (Langerhans) cells/mm². The observed pattern of cellular skin infiltration suggests an immune-mediated tissue injury including T-cell-mediated cytotoxicity and delayed hypersensitivity reactions in addition to direct parasitic action.     

The ultrastructure of the extracellular phase of bone as observed in frozen thin sections

Summary The fine structure of the extracellular phase of avian medullary bone and embryonic chick femur was examined in thin sections prepared by ultracryotomy and ultramicroincineration. Since contact with solutions was completely avoided, little or no loss or dislocation of mineral constituents could occur. Amorphous bone mineral (ABM) was present in two forms: as 15–30 nm spheres and as a structure-free haze. Removal of all organic material by low temperature ashing left the ABM intact. Crystals were usually associated with the ABM. In newly ossifying regions clusters or nodules of randomly oriented crystals and ABM appeared to coalesce when they reached approximately 1 m in diameter. In highly calcified regions crystals appeared to be oriented along collagen fibers. ABM did not appear to be associated with collagen. Unmineralized collagen was visible in osteoid after staining with dry OsO₄ vapor and it appeared to be diverted around nodules. Structures which resembled matrix vesicles were present. Selected area electron diffraction patterns indicated the presence of hydroxyapatite.     

Cancellous bone in the anterior part of the iliac crest

Summary Calcified tissue in the iliac crest and the adjoining ala of the ilium was investigated by scanning electron micrographs of thick, polished sections from which the marrow had been removed. Some quantitative properties of the trabeculae and of the marrow spaces were obtained from measurements on the images of the polished surfaces. Most of the cortex of the crest was porous, about 25% void, of varying thickness, intruding into the cancellous space in some regions. A structure containing about 35% by volume of bone was found at and near the anterior superior spine. Compact bone of normal appearance began as thin medial and proximal sheets below the crest, and thickened until at 20–30 mm it was substantial. The cancellous bone contained by these structures was varied. Two main zones were distinguished, whose junction ran from just below the anterior superior spine to the lower portion of the iliac fossa. In the lateral zone, adjacent to the crest, there were arch-like structures, commencing from the medial and proximal walls, and meeting, or even crossing, near the centre. The medial zone was distinguished by large marrow cavities and strongly orientated trabeculae. The relative volume of bone was similar in the two zones, falling from a maximum of 15–20% to about 5% in the regions of the anterior inferior spine and the iliac fossa. The average width of the trabeculae was significantly greater in the medial than in the lateral zone (W _b (m)0.16 mm, W_b(l)0.12 mm). Inclusions of very heavily constructed trabeculae, having average widths of about 0.35 mm, were found in both zones.     

Dynamic aspects of peripheral nerve changes in progressive neural muscular atrophy

Summary Serial nerve biopsies were performed at an early, and at an advanced stage of the disease in 2 patients with progressive neural muscular atrophy. The early biopsy showed a complete loss of the large diameter and thickly myelinated fibres, as well as an expansion of the endoneurial interstitium in both cases. Myelinated and unmyelinated fibres exhibited axonal degeneration in all biopsies occasionally. Onion bulb formation, a typical feature of peripheral neuropathy in neural muscular atrophy, was found to be prominent only in the latter biopsies. As regards the formal pathogenesis of hypertrophic neuropathy in neural muscular atrophy, axonal dystrophy and interstitial changes of the endoneurium were regarded as primary phenomena, demyelination and onion bulb formation as secondary. A possible causal relation between axonal dystrophy and interstitial changes, observed in these cases, is discussed in the light of the present literature.

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Zusammenfassung Bei 2 Patienten mit progressiver neuraler Muskelatrophie wurden Nervenbiopsien jeweils in einem frühen und in einem fortgeschrittenerem Stadium der Erkrankung entnommen und verglichen. In beiden Fällen zeigten bereits die frühen Biopsien ein völliges Fehlen der großkalibrigen, dickbemarkten Axone. Ebenfalls als frühe Veränderung wurde eine Erweiterung des endoneuralen Interstitiums festgestellt. Eine geringe Anzahl der vorhandenen bemarkten und unbemarkten Axone in allen Biopsien wies degenerative Veränderungen auf. Die für die progressive neurale Muskelatrophie typische Zwiebelschalenbildung der Schwannschen Zellen — möglicherweise eine Reaktion auf wiederholte De-und Remyelinisierungsvorgänge um dystrophische Axone — trat erst in den späteren Biopsien deutlicher hervor. Hinsichtlich der formalen Genese der hypertrophischen Neuropathie bei neuraler Muskelatrophie sind nach diesen Beobachtungen axonale Dystrophie und interstitielle Veränderungen des Endoneuriums als primäre Entmarkung und Zwiebelschalenbildung als sekundäre Phänomene zu betrachten. Die Möglichkeit einer kausalen Beziehung zwischen axonaler Dystrophie und interstitiellen Veränderungen wird an Hand der vorliegenden Befunde und Literatur diskutiert.

     

The effect of grinding of human dentine examined by X-ray diffraction, infrared spectroscopy, and electron microscopy

Analysis of human dentine by infrared spectrophotometry suggests that ball-grinding may result in damage of the apatite crystallites. The present study includes further assessments of this effect by X-ray diffraction, infrared spectroscopy, and electron microscopy. Each of three coarse-ground dentine samples (Group I) was divided into three portions of 30 mg. One of these portions was ball-ground for approximately 1 min (Group II), the second portion for 6 min (Group III), and the third portion for 23 min (Group IV). The 002 reflection showed line broadening, most marked from Group II to III. Electron microscopy showed a gradual change in crystallite appearance with increased grinding, most pronounced from Group II to III. These observations indicate that by prolonged grinding a limit is approached where no further changes in the crystallites occur. Electron microscopy also indicated that fracture of the crystallites might have occurred. This was probably accompanied by strains in the lattice. The infrared spectra indicated that no breakdown of the apatite structure had occurred during the entire grinding.     

肾恶性横纹肌样瘤临床病理分析及超微结构观察

目的:探讨肾恶性横纹肌样瘤(MRTK)的临床病理表现及超微结构特点。方法:用光镜、免疫组化及电镜等方法观察其病理组织学特点、免疫组化表达及超微结构。结果:光镜的典型特点是肿瘤细胞弥漫性排列,呈圆形、椭圆形及不规则形,胞质丰富,嗜酸性,部分胞质内可见透明状小体,细胞核圆形或椭圆形,核膜厚,核仁清楚,有的细胞核呈空泡状。部分核偏向一侧似浆细胞样。细胞内外见红染的包涵体。免疫组化显示肿瘤可表达多种抗原。电镜观察特异性表现是胞质内见中间丝及圆形不规则形纤维状或轮状小体。结论:肿瘤细胞弥漫排列呈浆细胞样,免疫组化多方向表达,可向神经性、上皮性、肌性等方向分化。电镜下胞质内中间丝及纤维轮状小体是特异性诊断标志。     

Electron microscopic localization of nitric oxide I synthase in the organ of Corti of the guinea pig

Nitric oxide synthase (NOS) activity has been detected previously in the mammalian cochlea at a light microscopic level. Here we present results of electron microscopic analysis for post-embedding immunoreactivity of neural-type NOS I in the cochlea of the guinea pig. Strong enzyme immunoreactivity was identified in the cytoplasm of inner and outer hair cells. Gold-labeled NOS I antibodies were mainly located in electron-dense areas of the cytoplasm, whereas electron-lucent regions of the receptor cells were nearly free from any immunoreactivity. In both types of hair cells anti-NOS I antibodies were also visible in the cuticular plates, hair bundles and nuclei. Further ultrastructural analysis revealed that the submembranous cisternae of the outer hair cells were nearly free from any reaction product, demonstrating that the whole cytoplasm of this hair cell was not immunoreactive. Other NOS I immunoreactivity was identified in the cuticular plates of the inner and outer pillar cells and in the cytoskeletal elements located in the apical parts of Deiter cells, forming the lamina reticularis or in cytoskeletal-containing regions in basal Deiter cells. Anti-NOS antibodies were visible in the nuclei of various cell types. Our findings suggest that nitric oxide produced by NO I synthase in the organ of Corti may act as a modulator of hair cell physiology during the processes of signal transduction with frequence selectivity.     

Use of electron spectroscopic imaging to determine element composition of the melanin granules in the stria vascularis of the guinea pig

Electron spectroscopic imaging (ESI) was used to analyze the element content of melanin granules in the stria vascularis seen in ultrathin sections of Spurr-embedded cochleae of the guinea pig. To determine element composition, ESI images were taken at different ionization edges, and non-specific background signals were subtracted digitally by an image processing system. The presence of calcium and nitrogen in the melanin granules could be demonstrated clearly. The calcium identified in the melanin granules was then compared with the spatial distributions of calcium binding sites after the application of an antimonate precipitation method, which was used to localize loosely bound calcium. Despite a high calcium concentration within the granules, only very small single scattered calcium precipitates could be detected between these structures as compared with the amount of calcium precipitates attached to the plasma membrane or located within the cell nuclei. The nearly complete absence of precipitates within the melanin granules after the application of antimonate suggests differences in calcium binding and mobility involved in various physiological processes of ion balance regulation within the stria vascularis. Received: 14 October 1997 / Accepted: 11 February 1998     

镫骨全切除和吸引噪声对豚鼠耳蜗功能与超微结构的影响

本实验应用耳蜗电图和扫描、透射电镜等技术观察了6组(44只)豚鼠镫骨全切除后,前庭窗开放时间的长短和听泡内使用吸引器对耳蜗功能和超微结构的影响程度.结果如下:①随前庭窗开放时间的延长、复合蜗神经动作电位(compound action potential.CAP)反应阈进行性提高,开放180min可导致底回部分外毛细胞变性或破坏;②镫骨全切除后,听泡内较长时间地(60min)应用吸引器可引起CAP明显阈移和耳蜗底回部分外毛细胞的不可逆病变;③镫骨未切除时,听泡内吸引60min,仅引起CAP阈移,但外毛细胞未见损伤.结果提示:术中长时间开放前庭窗和鼓室内应用吸引器可能是引起镫骨全切除术后感音神经性聋的因素之一,但鼓室成形术中鼓室内吸引噪声可能因其强度不够大,不足以引起术后感音神经性聋.