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1.
Little is known about the spinal cords of phylogenetically ancient actinopterygeans. The spinal cords of the chondrostean Acipenser schrenckii (Amur sturgeon), holostean Lepisosteus oculatus (spotted gar), and teleost Carassius auratus (goldfish) were, therefore, analyzed by immunohistochemistry, electron microscopy and two-dimensional gel electrophoresis. Morphology showed numerous similarities between sturgeons and gars. In both, a dorsal column between the two dorsal horns was lacking, giving the grey matter an inverted Y-shape. In goldfish, a small dorsal column was seen, the grey matter occupied a larger area, neuronal density was much higher, and a ventral commissure was apparent, which was absent in sturgeons and gars. In the white matter of sturgeons and gars, small caliber axons predominated, whereas larger axons were frequent in goldfish. Choline acetyltransferase immunoreactive neurons were prevalent in the ventral horns of all three fish, mainly in motoneurons, but stained fibers were only found in sturgeons and gars. gamma-aminobutyric acid positive cells were seen in both the ventral and the dorsal horns of all three fish. Distribution of serotonin (5-HT) and tyrosine hydroxylase (TH) immunoreaction was similar in sturgeons and gars, being located in both the ventral and the dorsal horns. In goldfish, 5-HT label was confined to the ventral horn and TH label was mainly observed in a cell group located ventromedially. Two-dimensional gel electrophoresis showed a gradual increase in protein number from sturgeons to gars to goldfish. In conclusion, the spinal cords of sturgeons and gars share many morphological and chemical features, distinguishing them from the goldfish spinal cord.  相似文献   

2.
The ultrastructural localization of putative excitatory (glutamate, aspartate) and inhibitory (taurine, glycine) amino acid neurotransmitters is described in several selected rat brain regions. In general, axon terminal profiles immunoreactive for excitatory amino acids formed asymmetric synapses with non-immunoreactive small diameter dendritic profiles or dendritic spines. In the cerebellum, both mossy fiber terminals and parallel fiber terminals were immunoreactive for glutamate and aspartate. In the hippocampus, mossy fiber terminals within the stratum lucidum of the CA3 region were immunoreactive for glutamate. Localization of glutamate and aspartate to cerebellar parallel and mossy fibers, as well as the identification of glutamate in hippocampal mossy fibers, is consistent with the excitatory nature of these fibers as described in previous physiological studies. Glutamate-like immunoreactive terminals were also identified in subnucleus caudalis of the spinal trigeminal nucleus and in the dorsal horn of the spinal cord. Immunoreactive axon terminals for two putative inhibitory neurotransmitters, glycine and taurine, displayed a greater number of morphological variations in synaptic structure. In the cerebellum, taurine-like immunoreactivity was present in both basket cell axon terminals which formed symmetric synapses with Purkinje cell neurons, and in a few mossy fiber terminals which formed asymmetric synapses with dendritic spines. In the area dentata of the hippocampus, taurine-like immunoreactive profiles formed asymmetric synapses with dendritic elements. Glycine-like immunoreactive terminals formed symmetric synapses with cell perikarya in both the ventral horn of the spinal cord and in the cochlear nuclei, and on axon terminals in the spinal trigeminal and cochlear nuclei. In contrast, some glycine-like immunoreactive terminals formed asymmetric synapses with distal dendritic profiles in the spinal cord and spinal trigeminal nucleus. The localization of taurine to cerebellar basket cell axons and glycine to axon terminals that synapse on ventral horn motor neuron perikarya is consistent with the hypothesis that these amino acids are functioning as inhibitory neurotransmitters at these synapses. Taurine localization to cerebellar mossy fibers and to fibers in the molecular layer of the dentate gyrus may be more consistent with a proposed neuromodulator role of taurine.  相似文献   

3.
The spinal cord is a vital link between the brain and the body and mainly comprises neurons, glial cells and nerve fibres. In this work, nonlinear optical (NLO) microscopy based on intrinsic tissue properties was employed to label‐freely analyze the cells and matrix in spinal cords at a molecular level. The high‐resolution and high‐contrast NLO images of unstained spinal cords demonstrate that NLO microscopy has the ability to show the microstructure of white and grey matter including ventral horn, intermediate area, dorsal horns, ventral column, lateral column and dorsal column. Neurons with various sizes were identified in grey matter by dark spots of nonfluorescent nuclei encircled by cytoplasm‐emitting two‐photon excited fluorescence signals. Nerve fibres and neuroglias were observed in white matter. Besides, the spinal arteries were clearly presented by NLO microscopy. Using spectral and morphological information, this technique was proved to be an effective tool for label‐freely imaging spinal cord tissues, based on endogenous signals in biological tissue. With future development, we foresee promising applications of the NLO technique for in vivo, real‐time assessment of spinal cord diseases or injures.  相似文献   

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6.
Using the transneuronal viral tracing method, the central nervous system (CNS) connections of the uterine horn were studied in virgin, pregnant, and in lactating rats. The frequency of viral labeling in the brain and the distribution of virus-infected neurons from the uterine horn were compared among groups. There was a marked difference in the frequency of viral labeling in the brain stem. In virgin rats more than half of the brain stems (5 out of 9) were labeled. In contrast, in pregnant animals viral-labeled neurons were detected in only a few cases (3 out of 16) and almost each brain stem of the lactating group was labeled (12 out of 13). A similar, less marked difference was observed in the hypothalamus. The pattern of distribution of infected neurons was similar in each group. In the brain stem, the nucleus of the solitary tract, dorsal motor nucleus of the vagus, area postrema, gigantocellular and paragigantocellular nucleus, ventrolateral medulla, A5 cell group, and caudal raphe nuclei were the most frequently labeled structures. In the diencephalon, viral-infected neurons were detected primarily in the hypothalamic paraventricular nucleus. The telencephalon was devoid of infected cells. Data suggest that the CNS control of the uterine horn varies depending on reproductive status. The low frequency of brain labeling in pregnant rats may be related to the almost complete lack of sympathetic fibers in the uterus prior to parturition and the very high frequency of labeling in lactating animals to the postpartum hyperinnervation of the uterus.  相似文献   

7.
Clinical and experimental grafting in Parkinson's disease has shown the need for enhanced survival of dopamine neurons to obtain improved functional recovery. In addition, it has been suggested that a limited number of surviving dopamine neurons project to the dopamine-denervated host striatum. The aim of this study was to investigate if subpopulations of ventral mesencephalic dopamine neurons project to their normal targets, i.e., dorsal vs. ventral striatum. Following implantation of human ventral mesencepahlic tissue into the lateral ventricle of dopamine-depleted rats, human-derived dopamine reinnervation was achieved both in dorsal and ventral striatum. Treatment with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) resulted in a degeneration of tyrosine hydroxylase (TH)-immunoreactive nerve fibers in dorsal striatum but not in ventral areas in some animals, while MPTP was without effect in other animals. TH-immunoreactive neurons were small and appeared shrunken in animals carrying grafts affected by the MPTP treatment. In conclusion, grafted dopamine neurons projected nerve fibers into areas that they normally innervate. Thus, when searching for factors that may enhance survival of grafted dopamine neurons it is important to study which subpopulation(s) of ventral mesencephalic dopamine neurons is affected, such that a proper reinnervation may be achieved.  相似文献   

8.
Gonadotropin releasing hormone (GnRH) is a peptide that is conserved in both vertebrate and invertebrate species. In this study, we have demonstrated the distribution pattern of two isoforms of GnRH‐like peptides in the neural ganglia and testis of reproductively mature male abalone, H. asinina, by immunohistochemistry and whole mount immunofluorescence. We found octopus (oct) GnRH and tunicate‐I (t) GnRH‐I immunoreactivities (ir) in type 1 neurosecretory cells (NS1) and they were expressed mostly within the ventral horn of the cerebral ganglion, whereas in pleuropedal ganglia they were localized primarily in the dorsal horn. Furthermore, tGnRH‐I‐ir were strongly detected in fibers at the caudal part of the cerebral ganglia and both ventral and dorsal horns of the pleuropedal ganglia. In the testis, only octGnRH‐ir was found primarily in the granulated cell and central capillaries within the trabeculae. These results suggest that multiple GnRH‐like peptides are present in the neural ganglia which could be the principal source of their production, whereas GnRH may also be synthesized locally in the testis and act as the paracrine control of testicular maturation. Microsc. Res. Tech. 77:110–119, 2014. © 2014 Wiley Periodicals, Inc.  相似文献   

9.
Based on current literature, the afferents of the superior olivary complex (SOC) are described including those from the cochlear nucleus, inferior colliculus, thalamus, and auditory cortex. Intrinsic SOC afferents and non-auditory afferents from the serotoninergic and noradrenergic systems are also described. New data are provided that show a differential distribution of serotoninergic afferents within the SOC: serotoninergic fibers were relatively sparse in the lateral and medial superior olives and the medial nucleus of the trapezoid body and were most numerous in periolivary regions. There are variations in the density of serotoninergic fibers within periolivary regions themselves. New data is also provided on auditory and non-auditory afferents to SOC neurons, which have known targets. These include: cochlear nucleus afferents to periolivary (lateral nucleus of the trapezoid body, LNTB) cells that project to the inferior colliculus; cortical afferents to periolivary (ventral nucleus of the trapezoid body, VNTB) cells that project to the cochlear nucleus; and serotoninergic and noradrenergic afferents to periolivary (LNTB and VNTB) cells that project to the cochlear nucleus. The relationships between other types of afferents and SOC neurons with known projections are also described as functional circuits. The circuits include those that are part of the ascending auditory system (to the inferior and superior colliculi, lateral lemniscus, and medial geniculate nucleus), the descending auditory system (to the cochlea and cochlear nucleus), and the middle ear reflex circuits.  相似文献   

10.
The regeneration of axons after a spinal cord injury or disease is attracting a significant amount of interest among researchers. Being able to assess these axons in terms of morphology, length and origin is essential to our understanding of the regeneration process. Recently, two specific axon tracers have gained much recognition; biotinylated dextran amine (BDA) 10 kDa as an anterograde tracer and cholera toxin‐B as a retrograde tracer. However, there are still several complexities when using these tracers, including the volume that should be administered and the best administration site so that a significant amount of axons are labeled in the area of interest. In this article, we describe some simple procedures for injecting the tracers and detecting them. We also quantified the number of axons at different locations of the spinal cord. Our results show axons labeled from motor cortex injections traveled down to the lumbosacral spinal cord in 2 weeks, while BDA injections into the lateral vestibular nucleus and reticular formation took 3 weeks to label axons in the lumbosacral spinal cord. Moreover, this protocol outlines some basic procedures that could be used in any laboratory and gives insight into the number of axons labeled and how procedures could be tailored to meet specific researcher's needs. Microsc. Res. Tech. 76:1240–1249, 2013. © 2013 Wiley Periodicals, Inc.  相似文献   

11.
Immunohistochemistry is used to demonstrate histamine-immunoreactivity in the CNS of spiders. We found histamine-immunoreactivity in the photoreceptors of different spiders. Therefore, we suggest that histamine is a neurotransmitter of photoreceptors in all arthropods, since it is also known to occur in the photoreceptors of the other main arthropod taxa (Merostomata, Crustacea, and Insecta). We also describe a system of only six omnisegmental histamine-immunoreactive neurons within the central nervous system. These histamine-immunoreactive neurons can be divided into two subgroups: a dorsal system with two cells per hemisphere and a ventral system with only one cell per hemisphere. All six cells have extended arborizations in both the motor and the sensory areas of all neuromeres in the suboesophageal ganglionic mass. In contrast to araneomorph spiders, two additional sets of histamine-immunoreactive neurons were detected in mygalomorph spiders. The first set consists of seventeen cells with their cell bodies located in the cheliceral ganglion and projecting to central areas of the protocerebrum. The second set contains many if not all sensory projections from the tarsal organs on all eight legs and the pedipalps to the Blumenthal neuropil.  相似文献   

12.
The extrabulbar olfactory projections (EBOP) is a collection of nerve fibers that originate from primary olfactory receptor neurons. These fibers penetrate into the brain, bypassing the olfactory bulbs (OBs). While the presence of an EBOP has been well established in teleosts, here we morphologically characterize the EBOP structure in four species each with a different morphological relationship of OB with the ventral telencephalic area. Tract‐tracing methods (carbocyanine DiI/DIA and biocytin) were used. FMRFamide immunoreactive nervus terminalis (NT) components were also visualized to define any neuroanatomical relationship between the NT and EBOP. Unilateral DiI/DiA application to the olfactory chamber stained the entire olfactory epithelium, olfactory nerve fibers, and ipsilateral olfactory bulb. Labeled primary olfactory fibers running ventromedially as extrabulbar primary olfactory projections reached various regions of the secondary prosencephalon. Only in Moenkhausia sanctaefilomenae (no olfactory peduncle) did lipophilic tracer‐labeled fibers reach the ipsilateral mesencephalon. The combination of tracing techniques and FMRFamide immunohistochemistry revealed a substantial overlap of the label along the olfactory pathways as well as in the anterior secondary prosencephalon. However, FMRFamide immunoreactivity was never colocalized in the same cellular or fiber component as visualized using tracer molecules. Our results showed a certain uniformity in the neuroanatomy and extension of EBOP in all four species, independent of the pedunculate feature of the OBs. The present study also provided additional evidence to support the view that EBOP and FMRFamide immunoreactive components of the NT are separate anatomical entities. Microsc. Res. Tech. 78:268–276, 2015. © 2015 Wiley Periodicals, Inc.  相似文献   

13.
This review presents information about multiple neurochemical substances in the carotid body. Nerve fibers around blood vessels and glomus cells within the chemoreceptive organ contain immunoreactivities (IR) for tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), substance P (SP), galanin (GAL), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), calretinin (CR), calbindin D-28k (CB), parvalbumin (PV), and nitric oxide synthase (NOS). Parasympathetic neurons scattered around the carotid body contain VIP, choline acetyltransferase, and vanilloid receptor 1-like receptor. In the mammalian carotid body, transection of the carotid sinus nerve (CSN) causes the absence or decrease of CGRP-, SP-, and NOS-immunoreactive (IR) nerve fibers, whereas all NPY-IR nerve fibers disappear after removal of the superior cervical ganglion. Most VIP-IR nerve fibers disappear but a few persist after sympathetic ganglionectomy. In addition, the CSN transection appears to cause the acquisition of GAL-IR in originally immunonegative glomus cells and nerve fibers within the rat carotid body. On the other hand, 4%, 25%, 17%, and less than 1% of petrosal neurons retrogradely labeled from the rat CSN contain TH-, CGRP-, SP-, and VIP-IR, respectively. In the chicken carotid body, many CGRP- and SP-IR nerve fibers disappear after vagus nerve transection or nodose ganglionectomy. GAL-, NPY-, and VIP-IR nerve fibers mostly disappear after removal of the 14th cervical ganglion of the sympathetic trunk. The origin and functional significance of the various neurochemical substances present in the carotid body is discussed.  相似文献   

14.
This study aimed to evaluate the integration of transplanted choroidal plexus epithelial cells with organotypic spinal cord slices. Organotypic spinal cord slices, normally cultured for 6 days, were divided into control group (Ctrl) and transplanted group (T). The choroidal plexus epithelial cells were dissociated and primary cultured (C group). The choroidal plexus epithelial cells cultured for 6–7 days were labeled by 1,1’-dioctadecyl-3,3,3’,3’-tetramethyl-indocarbocyanineperchlorate (CM-Dil), and were identified by transthyretin (TTR) in immunocytochemistry. They were adjusted to the density of 0.5–1 × 107/ml, then 2 μl cells suspension were transplanted to the spinal cord slices in the T group. The same amount of basal medium was dripped on the spinal cord slices in the Ctrl group. After 14 days of transplantation, the differentiations into neurons and astrocytes, and the synapses were identified by immunofluorescence histochemistry. At the same time, the ratios of cell differentiations and synapses in new system, and the changes of MAPK signaling pathway were tested by western blotting. The choroid plexus epithelial cells were well labeled by CM-Dil and were immune-stained by TTR in immunocytochemistry. The choroid plexus epithelial cells bodies were small when transplanted on the spinal cord slices, but big when transplanted on the polyester membrane inserts. The transplanted cells could differentiate into astrocytes, and possibly differentiate into neurons, and there were a large number of synaptophysin positive vesicles between transplanted cells and organotypic spinal cord slices in immunofluorescence histochemistry. The levels of GFAP, TUB-III and synaptophysin in the T group were higher than which in the Ctrl and C groups in western blotting (P < 0.05). And the ratios of p-JNK/JNK and p-P38/P38 in the T group were significantly lower than which in the Ctrl and C groups (P < 0.05). But the ratio of p-ERK/ERK in the three groups was of no significant difference. The transplanted choroidal plexus epithelial cells can integrate with organotypic spinal cord slices into a new system.  相似文献   

15.
Locusts are attractive model preparations for cellular investigations of neurodevelopment. In this study, we investigate the immunocytochemical localization of histamine in the developing ventral nerve cord of two locust species, Schistocerca gregaria and Locusta migratoria. Histamine is the fast neurotransmitter of photoreceptor neurons in the compound eye of insects, but it is also synthesized in interneurons of the central nervous system. In the locust ventral nerve cord, the pattern of histamine-immunoreactive neurons follows a relatively simple bauplan. The histaminergic system comprises a set of single, ascending projection neurons that are segmentally arranged in almost every neuromere. The neurons send out their axons anteriorly, forming branches and varicosities throughout the adjacent ganglia. In the suboesophageal ganglion, the cell bodies lie in a posteriolateral position. The prothoracic ganglion lacks histaminergic neurons. In the posterior ganglia of the ventral nerve cord, the somata of the histaminergic neurons are ventromedially positioned. Histamine-immunoreactivity starts around 50% of embryonic development in interneurons of the brain. Subsequently, the neurons of the more posterior ganglia of the ventral nerve cord become immunoreactive. From 60% embryonic development, the pattern of soma staining in the nerve cord appears mature. Around 65% of embryonic development, the photoreceptor cells show histamine-immunoreactivity. The histaminergic innervation of the neuropile develops from the central branches toward the periphery of the ganglia and is completed right before hatching.  相似文献   

16.
Retrograde tracing with True Blue was combined with immunocytochemistry to determine the source of any calretinin-immunoreactive (CR-ir) nerves projecting to the rat ovary. In the ovary, a strong signal for calretinin immunoreactivity was localized in interstitial gland cells; however, no intraovarian CR-ir nerves could be demonstrated. When the superior ovarian nerve was isolated, cut, and True Blue applied to the proximal end, the fluorescent dye was retrogradely transported to a population of cells located in T-12, T-13, and L-1 dorsal root and paravertebral ganglia. There was virtually no dual labeling of cells in these ganglia with calretinin (< 0.009% dual labeling in dorsal root and <0.014% in paravertebral ganglia). However, greater than two-thirds of the True Blue-labeled cells were immediately adjacent to CR-ir cells in dorsal root ganglia. This arrangement is suggestive of a paracrine mechanism between CR-ir cells and cells projecting to the ovary. In paravertebral ganglia, 63% of cells projecting to the ovary were surrounded completely or partially by beaded CR-ir nerve fibers. The source of these fibers (sensory or preganglionic sympathetic) is unknown but hypothesized to be preganglionic. Collectively, these observations suggest a participatory role for calretinin in ovarian function, either directly via effects on the interstitial gland or indirectly by influencing neurons projecting to the ovary.  相似文献   

17.
In the cattle, SCO-spondin was shown to be a brain-secreted glycoprotein specifically expressed in the subcommissural organ (SCO), an ependymal differentiation located in the roof of the Sylvian aqueduct. Furthermore, SCO-spondin makes part of Reissner's fiber (RF), a structure present in the central canal of the spinal cord. Sequencing of overlaping cDNA inserts after successive screening of a cattle SCO cDNA expression library allowed characterization of the complete sequence of this novel protein. Conserved domains were identified including twenty-six thrombospondin type 1 repeats (TSRs), nine low-density lipoprotein receptor LDLr type A domains (LDLRA), two epidermal growth factor EGF-like domains, and homologies to mucins and the von Willebrand factor were found in the amino- and carboxy- termini. In addition, SCO-spondin shows a unique arrangement "in mosaic" of these domains. The putative function of SCO-spondin in neuronal differentiation is discussed regarding these features and homologies with other developmental molecules of the central nervous system exhibiting TSR domains, and involved in axonal guidance.To correlate molecular and functional features of SCO-spondin, we tested the effect of oligopeptides whose sequences include highly conserved regions of the TSRs, LDLRA repeats, and a potent site of attachment to glycosaminoglycan, on cortical and spinal cord neurons in primary cell cultures. Peptides corresponding to SCO-spondin TSRs markedly increased adhesivity and neuritic outgrowth of cortical neurons and induced disaggregation of spinal cord neurons. Thus, SCO-spondin is a candidate to interfere with neuronal development and/or axonal guidance during ontogenesis of the central nervous system in modulating side-to-side and side-to-substratum interactions, and in promoting neuritic outgrowth. RF proper has a wide range of activity on neuronal differentiation, including survival, aggregation, and disaggregation effects and neurite extension of cortical and spinal cord neurones "in vitro." Thus, the SCO/RF complex may interact with developmental processes of the central nervous system including the posterior commissure and spinal cord differentiation.  相似文献   

18.
The use of antisera directed against conjugates of histamine and serotonin has revealed the locations of neurons labeling for these transmitters in the nervous system of barnacles. Photoreceptors label for histamine but not serotonin and also satisfy a number of other criteria indicating that histamine is their neurotransmitter. Photoreceptors also take up radioactively labeled histamine but not serotonin. Within the barnacle's brain no somata are consistently found that label with antiserum against histamine, but one to three pairs of small cells, depending on species, label with antiserum against serotonin. The most impressive serotonin-like immunoreactivity in the brain, however, is in a pair of large fibers ascending through the circumesophageal connectives and ramifying extensively. Within the ventral ganglion, the only other ganglion in the barnacle, ten pairs of cells label with antiserum against histamine. These neurons are confined to the posterior portion of the ganglion but ramify extensively throughout the ganglion. Antiserum against serotonin labels about 15 cell pairs, depending on species, located throughout the ganglion. The positions of the arbors of many of these cells suggest that these amines have a role in modulating either the motor pathways underlying feeding or the visual pathways responsible for the detection of shadows.  相似文献   

19.
胆碱乙酰转移酶(choline acetyltransferase, CHAT)是催化乙酰胆碱(acetylcholine,Ach)合成的特异性酶,是监测中枢和周围神经系统中胆碱能神经元功能状态的最特异性指标。本实验应用激光扫描公共聚焦显微镜、免疫荧光纳米金标记技术和包埋前免疫标记电镜技术,观察ChAT在脊髓前角运动神经元中超微结构水平上的分布。包埋前免疫标记降低了电镜制样对抗原性的破坏;激光扫描共聚焦显微镜荧光图像为透射电镜超薄切片提供了精确定位支持,有利于选择免疫标记效果更好的细胞进行超薄切片;振动切片有利于结构及抗原的保存使纳米金颗粒更易渗透入细胞,提高了免疫标记阳性率。  相似文献   

20.
Trk neurotrophin receptors are transmembrane tyrosine kinase proteins known as TrkA, TrkB, and TrkC. TrkA is the high affinity receptor for nerve growth factor, TrkB is the one for both brain-derived neurotrophic factor and neurotrophin-4, and TrkC is the preferred receptor for neurotrophin-3. In the adult mammalian brain, neurotrophins are important regulators of neuronal function and plasticity. This study is based on Nothobranchius furzeri, a teleost fish that is becoming an ideal candidate as animal model for aging studies because its life expectancy in captivity is of just 3 months. In adult N. furzeri, all three investigated neurotrophin Trk receptors were immunohistochemically detected in each brain region. TrkA positive neuronal perikarya were localized in the dorsal and ventral areas of the telencephalon and in the cortical nucleus; TrkB immunoreactivity was observed in neuronal perikarya of the dorsal and ventral areas of the telencephalon, the diffuse inferior lobe of the hypothalamus, and Purkinje cells; TrkC positive neuronal perikarya were detected in the most aboral region of the telencephalon, in the magnocellular preoptic nucleus and in few neurons dispersed in the hypothalamus. Numerous positive fibers were widely distributed throughout the brain. Radial glial cells lining the mesencephalic and rhombencephalic ventricles showed immunoreactivity to all three Trks. These findings suggest an involvement of neurotrophins in many aspects of biology of adult N. furzeri.  相似文献   

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