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Notes: Abstract. Objectives and Methods: A fluorescence and electron microscopical approach, based on the transection of the rat optic nerve and the axotomy-induced transcellular labelling of activated retinal microglial cells, using the carbocyanine dye 4Di-10ASP, was employed to monitor phagocytosis in the injured central nervous system. After survival times ranging between two days up to three months, retinal flat-mounts were inspected and photoconverted.¶Results: Fluorescence microscopy revealed that within a few days microglia became transcellularly stained due to the phagocytosed 4Di-10ASP-labelled neuronal debris. Ultrastructural analysis confirmed that marked ganglion cell-derived material was incorporated into phagosomes of various sizes. Though immediate phagocytic intake was not observed, the nature of the detected phagosomes suggests that small fractions of degenerated neurons are incorporated.¶Conclusions: The approach presented, utilizing function-dependent transcellular fluorescent labelling of phagocytic microglia, might enrich further experimental studies of glia-neuron interactions in the injured nervous system.

Notes: Abstract  Extracellular matrix proteoglycans accumulated in perineuronal nets and in certain neuropil zones have been shown to influence the immediate neuronal microenvironment, and to contribute to the chemoarchitectonic characteristics of neuronal networks. Studies in different placental mammals, including the human, have suggested that the major principles of extracellular matrix distribution remained constant during phylogenesis of the different mammalian strains. However, the comparison of matrix distribution between various species also indicates that striking deviations from the basic pattern may occur, although their functional significance appears unknown as yet. This study examines the extracellular matrix in the forebrain of a basic American marsupial, which has evolved independently of placental mammals for more than 100 million years. Brain sections obtained from adult gray short-tailed opossums (Monodelphis domestica) were stained for extracellular matrix components using the N-acetylgalactosamine-binding lectin Wisteria floribunda agglutinin (WFA), a polyclonal antibody against chondroitin sulfate proteoglycans (CSPG), and biotinylated hyaluronectin for the detection of hyaluronan. In subcortical regions, the distribution patterns of WFA-stained and CSPG-immunoreactive perineuronal nets were similar to those reported previously in placental mammals. In contrast, a unique distribution was found in the neocortex. This distribution was characterized by the presence of perineuronal nets around pyramidal cells and matrix components within the adjacent neuropil that together form a continuously labeled zone in layer V. Weakly stained nets ensheathed less numerous pyramidal cells in the upper layers II/III and a few multipolar cortical neurons. Dual staining experiments showed that cortical net-associated neurons were rarely immunopositive for parvalbumin. This fact, in addition to the predominant association of extracellular matrix components with layer V pyramidal cells, differentiates the neocortex in Monodelphis from that of all placental mammals studied to date. Regarding the basic phylogenetic position of this marsupial species it remains to be shown if these distribution characteristics of extracellular matrix may represent also a basic feature of cortical organization.

Notes: Abstract A 7-year-old boy had clinical features of metachromatic leucodystrophy (MLD), however, an increased urinary sulphatide excretion was found in the presence of normal arylsulphatase A (and α-galactosidase A) activity. A rectal biopsy showed metachromatically staining storage macrophages as well as nonmetachromatic, but PAS-positive, submucosal neurons filled with membranous cytoplasmic bodies. These two types of storage material led to testing for a sphingolipid activator protein (SAP) deficiency. Loading tests with sulphatide and globotriaosylceramide showed deficient turnover of both sphingolipids in cultured fibroblasts. Using the Ouchterlony method, there was no reactivity between a described anti-SAP 1 antiserum and the patient's fibroblast extracts. This new case of SAP-1 deficient MLD was compared with the four cases of this variant known from the literature. Our results indicate that rectal biopsy morphology and lipid loading biochemistry should prove useful for the screening of SAP defects.

Notes: We should know the effects of soil use and management on the contents and forms of soil phosphorus (P) and the resulting potential for leaching losses of P to prevent eutrophication of surface water. We determined P test values, amounts of sequentially extracted forms of P, P sorption capacities and degrees of P saturation in 20 differently treated soils and compared these data with leaching losses in lysimeters. One-way analyses of variance indicated that most fractions of P were significantly influenced by soil texture, land use (grassland, arable or fallow or reafforestation), mineral fertilization and intensity of soil management. Generally, sandy soils under grass and given large amounts of P fertilizer contained the most labile P and showed the largest P test values. Fallow and reafforestation led to smallest labile P fractions and relative increases of P extractable by H2SO4 and residual P. Arable soils with organic and mineral P fertilization given to crop rotations had the largest amounts of total P, labile P fractions and P test values. The mean annual concentrations of P in the lysimeter leachates varied from 0 to 0.81 mg l–1 (mean 0.16 mg l–1) and the corresponding leaching losses of P from 〈 0.01 to 3.2 kg ha–1 year–1 (mean 0.3 kg P ha–1 year–1). These two sets of data were correlated and a significant exponential function (R2 = 0.676) described this relation. Different soil textures, land uses and management practices resulted in similar values for P leaching losses as those for the amounts of labile P fractions. Surprisingly, larger rates of mineral P fertilizer did not necessarily result in greater leaching losses. The contents of P extracted by NaHCO3 and acid oxalate and the degrees of P saturation were positively correlated with the concentrations of P in leachates and leaching losses. As the P sorption capacity and degree of P saturation predicted leaching losses of P better than did routinely determined soil P tests, they possibly can be developed as novel P tests that meet the requirements of plant nutrition and of water protection.

Notes: Borna disease virus (BDV) causes a non-purulent meningoencephalitis and retinitis in a variety of species. In Lewis rats infected intracerebrally with the highly neurotropic BDV the retina is one of the most severely affected central nervous system (CNS) structures. While BDV-induced damage in the brain has previously been shown to be caused by a T-cell-dependent process, the immunopathological mechanisms leading to BDV-induced retinitis, remain to be elucidated. RNA samples from retinae were subjected to RNase protection assays to detect transcripts of proinflammatory cytokines and chemokines known to be involved in the recruitment of T-cells and macrophages in the CNS. The observed expression profile of proinflammatory cytokines and chemokines, as well as the immunohistochemical detection of αβTCR-positive and CD8-positive T-cells in the BDV-infected retinae, is reminiscent of the situation observed in the brains of Lewis rats during the acute phase of Borna disease. This suggests, that similar immunopathological mechanisms are operating in retinae and brains of infected rats. This research was supported by grants from the Deutsche Forschungsgemeinschaft (J.S.-PA 615/1-1).

Notes: We investigated the embryotoxicity of ozone (O3), after gas exposure in ovo, to chick embryos of the Lohmann-Weiss strain. The main aim of the study is to reduce the microbiological content, particularly of Salmonella enteritides on the egg surface by the use of ozone in different concentrations and response times. The aim of our morphological study is to analyse the influence of gas exposure to the embryonal development of chick. Ozone has proved its worth as a disinfectant without the harmful adverse effects of formaldehyde. The chick embryo provides a useful and well-defined model for analysis of toxicity of chemicals, particularly with regard to an alternative disinfectant: ozone. Embryotoxicity of ozone in developing chick embryos was studied by examining different parameters: deformities and pathohistology of heart, liver, kidney and spleen, weight, length and survival of embryo. After oviposition the eggs were transferred to a chamber filled with ozon-oxygen-mixture in a defined concentration (1% - 5.22%). After achievement of the ozone concentration the eggs were incubated over various response times have 20, 30, 60 min or 17 h. After the incubator times of 18, 19 or 20 days we studied the development of the gas exposed embryos and unexposed controls. The organs were fixed in 4% formalin, embedded in paraffin, cut and stained with hematoxylin-eosin. High ozone concentration over 5% and high response time of 1 hour provided low graded survival rates of 3.3–13.3%. There was significantly increase of the survival rate after decline of ozone concentration and response time of 90–100%. We found also a remarkable decrease of weight and length in fewer surviving gas exposed embryos with high ozone concentration and response time (Fig. 1.), compared with the normal development of gas-exposed embryos with study of high survival rate (Fig. 2.). We calculated a 60% increase in the survival rate of embryos exposed to high ozone concentration without response time. In low ozone concentration (1.8%) with high response time of 17 hours the survival rate decreased to 10%. We found no signs of deformities and pathohistology of organs in our embryological examination. In conclusion, ozone has a dose-dependent effect. The response time constitutes an important factor in the survival and growth of chick embryos after exposure to ozone in ovo. 〈displayedItem type="text" xml:id="fu3"〉1〈mediaResource alt="image" href="urn:x-wiley:03402096:AHE669_44_44:AHE_669_fu3"/〉 [ High ozone concentration over 5% and high response time of 1 hour result in a low survival rate of 3,3% and the surviving embryo shows a decrease in body weight and length (right) in comparison to the biggest (left) and lowest (middle) unexposed embryo. ] 〈displayedItem type="text" xml:id="fu4"〉2〈mediaResource alt="image" href="urn:x-wiley:03402096:AHE669_44_44:AHE_669_fu4"/〉 [ Height comparison between the unexposed controls (both embryos on the left side) with gas exposed embryos (both embryos on the right side) in a study with low ozone concentration of 1% and low response time of 20 minutes. There are no differences in body weight and length between the unexposed and exposed embryos. ]

Notes: This combined light and scanning electron microscopic (SEM) analyses are the first investigation of the fine morphology of the fetal membranes of the placenta in the Asian elephant. We used two term placentas with gestation periods of 634 days (Zoological Garden Leipzig) and 657 days (Zoo Hagenbeck) and selected unequal sites as specimens, particularly - free membrane of the allantois, - lateral edge of the placenta band, the ‘haemophagous zone’, - allantochorion near the placental band, and allantochorion on the end of the chorioallantoic sac. Microscopically the free membrane of the allantois shows a simple, cuboidal epithelium with apical domains of microplicae and microvilli (fig. 1, inset). In the SEM analyses we documented a cobblestone-like architecture of the epithelial cells with various cell sizes, small, middle and giant (fig. 1.). Furthermore, we found pear-shaped cells with long pedicle, attached to the basement membrane and cell openings, presumably due to cell desquamation (fig. 2.). The haemophagous zone was characterized with a lamellate system between the long finger-like chorionic villi and the maternal blood lacunae. The simple, squamous and columnar trophoblast cells are here bathed in the maternal blood. In some chorionic villi brown-yellow material is deposited in the fetal connective tissue. A special result was the presence of strip-like microplicae in the middle of chorionic villi. The allantochorion near the placental band sometimes indicated simple and ramified chorionic villi with smooth, gyrus-like trophoblast cells and foamy cytoplasm. The attached allantois possesses a simple, columnar epithelium with microvilli. Furthermore, we observed villi-like projections rising up to the allantoic cavity. The white ‘pustules’ on the outer surface of the end of the chorioallantoic sac consist of stratified squamous epithelia. Numerous mitoses were documentedin the basal domain of the pustules. A surprisingly result was the presence of intra-epithelial capillaries. They invade the trophoblast cells to presume a nutrient exchange also outside of the placental band with its important functional metabolism. This study revealed unknown and interesting features of the epithelial layer of different locations in the placenta of the Asian elephant. Perhaps, with a detailed knowledge of the morphology of the epithelium we could convey a better understanding of functional cohesions of elephant placentation. 〈displayedItem type="text" xml:id="fu1"〉1〈mediaResource alt="image" href="urn:x-wiley:03402096:AHE669_43_43:AHE_669_fu1"/〉[ The free membranes of the allantois of the term placenta of the Asian elephant (Elephas maximus) shows a cobblestone-like architecture of the simple cuboidal epithelial cells with various sizes. The surface morphology was determined through microplicae and microvilli. ] 〈displayedItem type="text" xml:id="fu2"〉2〈mediaResource alt="image" href="urn:x-wiley:03402096:AHE669_43_43:AHE_669_fu2"/〉 [ Numerous pear-shaped cells with long pedicle exist among the cuboidal epithelial cells, attached on the basement membrane. The cells possessed modifications of their surface, particularly microplicae. Some pear-shaped cells shows only the pedicle, perhaps that is a loss through preparation or its presumable due to cell desquamation. One erythrocyte is localized between these cells. ]

Notes: Summary Intravenous digital subtraction angiography (IVDSA) was used to evaluate 44 patients with suspected otolaryngologic abnormalities. Sixteen had IVDSA for pulsatile tinnitus or suspected glomus tumor of the petrous bone. Nine patients were evaluated because of pulsatile neck masses, and 12 others had suspected tumors of the neck, face, and paranasal sinuses. Seven had IVDSA following head and neck trauma. The technique of examination is described. The current indications of IVDSA in head and neck radiology are discussed. It is concluded that IVDSA is a suitable substitute for conventional angiography for many otolaryngologic conditions and, because of its safety, can be used more liberally.

Notes: Summary Ossification of the posterior longitudinal ligament is a potentially curable cause of thoracic myelopathy. As this case illustrates, MRI can confirm the diagnosis, demonstrate other concomitant pathology such as disc herniation, and help assess the clinical significance of OPLL by showing spinal word compression.