Immunohistochemical analysis of development of desmin-positive hepatic stellate cells in mouse liver

Development of desmin-positive hepatic stellate cells was studied in mice using double immunofluorescent techniques and in vitro cultures with special attention given to their cell lineages. Several studies recently reported on the presence of cells that are immunologically reactive with both antidesmin and anticytokeratin antibodies in young fetal rat livers, and suggested the possibility that these cells give rise to hepatocytes and hepatic stellate cells. At early stages of mouse liver development, stellate cells with desmin-positive filaments were scattered in the liver parenchyma. However, the stellate cells definitely differed from hepatoblasts and hepatocytes in terms of their morphology and expression of desmin and hepatoblast and hepatocyte-specific E-cadherin in the liver. Fetal hepatoblasts and hepatocytes did not react with antidesmin antibodies, nor did desmin-positive stellate cells express E-cadherin in vivo and in vitro. Thus it is likely that desmin-positive stellate cells and hepatoblasts belong to different cell lineages. In the fetal liver, the desmin-positive stellate cells surrounded blood vessels, and extended their processes to haematopoietic cells and megakaryocytes. Many, but not all, hepatoblasts and hepatocytes were observed to be associated with the stellate cells. At fetal stages, cellular processes positive for desmin in the stellate cells were also thick compared with those in the adult liver, in which desmin-positive stellate cells lay in Disse's space and were closely associated with all hepatocytes. These developmental changes in the geography of desmin-positive cells in the liver parenchyma and their morphology may be associated with their maturation and interactions with other cell types.     

Changing distribution patterns of synaptophysin‐immunoreactive structures in the human dorsal striatum of the fetal brain

Within the striatum two compartments, matrix and patches, can be distinguished by differences in the expression of neuroactive substances, afferent and efferent connections and time of neurogenesis. The present study was done to demonstrate the pattern of synaptophysin (SYN) expression which is indicative of synaptogenesis in the human fetal striatum (15th–32nd weeks of gestation) with special reference to developmental changes. From the 15th to the 22nd gestational weeks an intense diffuse SYN immunolabelling of striatal patches is observed. In the matrix SYN‐immunoreactive fiber bundles are seen until the 20th week. Thereafter, the matrix is nearly devoid of SYN‐immunoreactive structures. From the 28th week of gestation the matrix contains diffuse SYN immunoreactivity which gradually becomes as intense as that of the patches. The latter, thus, can no longer be delineated in the 30th week. The results show that fibrous SYN immunolabelling most probably indicating intra‐axonal transport of synaptic vesicles can only be observed during the first half of gestation. Moreover, it becomes obvious that the patch compartment can selectively be visualized by anti‐SYN until the 28th week. This pattern may correspond to the early dopaminergic innervation from the substantia nigra which is known to reach the developing patches. From the 28th week a transition from patchy to diffuse immunolabelling is seen. The increase in matrix labelling may be due to the occurrence of new neuronal contacts. The changeover from patchy to homogeneous SYN immunolabelling takes place distinctly earlier than changes in the distribution of other neuroactive substances described before. Anat Rec 258:198–209, 2000. © 2000 Wiley‐Liss, Inc.     

Fine-needle aspiration cytology of malignant retroperitoneal paraganglioma

The cytologic findings in a fine-needle aspiration sample are described from a large retroperitoneal mass in a 56-yr-old male. The aspiration was performed under CT guidance using a 22-gauge needle, maintaining negative pressure. For cytologic study, on-site smears were prepared and stained by the Papanicolaou method. Additionally, cytospin, filter preparations, cell blocks from the aspirate, histology, and electron microscopy of the tumor tissue were performed. Also, immunodiagnostic staining for neuron-specific enolase (NSE), chromogranin, S-100 protein, vimentin, HMB45, cytokeratin, and Grimelius preparation was performed on cytologic and histologic material. The cytologic material was characterized by cords, suggestive acinar structures, and small follicle-like clusters of cells, while cell blocks from the aspirate showed oval or spindle-shaped nuclei with a somewhat fasciculated appearance. However, sections from the tumor tissue showed an organoid “zellballen” pattern. The nuclei were round, oval, spindly, and epithelioid, with moderate to scanty cytoplasm, smooth nuclear membranes, fine, evenly dispersed chromatin, slight hyperchromasia, and mild pleomorphism. No intranuclear vacuoles were seen. Positive immunohistochemical staining for NSE, chromogranin, and Grimelius preparation was noted in the tumor cells, while staining for vimentin, HMB45, and cytokeratin was negative. Electron microscopy of the tumor tissue revealed the presence of variable numbers of round, membrane-bound, electron-dense neurosecretory granules. The cytohistologic and ultrastructural findings are presented, as well as the results of immunodiagnostic staining which helped in the diagnosis of retroperitoneal paraganglioma, an infrequently reported tumor. Diagn. Cytopathol. 1998;18:287–290. © 1998 Wiley-Liss, Inc.     

Responses of cortical noradrenergic and somatostinergic fibres and terminals to adjacent strokes and subsequent treatment with NGF and/or the ganglioside GM1

The occurrence of sprouting by fibre systems in the neocortex following lesion is still a controversial issue. In previous studies, we showed a nerve growth factor (NGF)-induced sprouting and hypertrophy of presynaptic terminals in the cholinergic fibres of the rat neocortex following stroke-type lesions, effects that were potentiated by the monosialoganglioside GM1. The present study investigated whether exogenous NGF and/or GM1 treatment could also affect the noradrenergic and somatostinergic systems in the neocortex. Immediately following unilateral vascular decortication, adult rats received, via minipump, a 7-day infusion of vehicle, NGF (12 μg/day) and/or GM1 (1.5 mg/day) into the cerebroventricular space. Thirty days postlesion, the animals were perfused with histological fixatives, the brains were removed, and relevant sections were processed for dopamine β-hydroxylase and somatostatin immunocytochemistry at the light and electron microscopic levels. A Quantimet 920 image analysis system was used for the quantification of fibre length and size of presynaptic boutons. The lesion caused a reduction in the dopamine β-hydroxylase-immunoreactive fibre length, which was not attenuated by either NGF or GM1 treatment or both. The somatostatin-immunoreactive network, in contrast, was unaffected by the lesion, and there was no sprouting of somatostatin fibres following trophic factor therapy. We also found no significant differences in the size and number of synapses of both the dopamine β-hydroxylase-immunoreactive and somatostatin-immunoreactive boutons following lesion and drug treatments. These results indicate that NGF and/or GM1 therapies do not cause regrowth in the noradrenergic and somatostatinergic cortical fibre networks or their presynaptic elements following a cortical devascularizing lesion. J. Neurosci. Res. 50:627–642, 1997. © 1997 Wiley-Liss, Inc.     

Pure alveolar rhabdomyosarcoma of the uterine corpus

Herein is presented a very rare case of alveolar rhabdomyosarcoma in the uterine corpus of a 72-year-old woman. The wall of the uterine corpus was replaced by multiple whitish-yellow, friable nodules, measuring up to 6 cm. Microscopically, the tumor was predominantly composed of round to polygonal cells arranged in an alveolar, papillary or nest pattern intermingled with multinuclear giant cells with abundant eosinophilic cytoplasm. Extensive sampling failed to show epithelial elements. Immunohistochemically, the tumor was positive for striated muscle markers such as myoglobin, myoD1 and myogenin. Metastatic lesions were found in the retroperitoneum and pelvic lymph nodes. The patient was treated by postoperative chemotherapy, but she died of systemic metastases 12 months after surgery.     

Morphological abnormalities of embryonic cranial nerves after in utero exposure to valproic acid: implications for the pathogenesis of autism with multiple developmental anomalies

Autism is often associated with multiple developmental anomalies including asymmetric facial palsy. In order to establish the etiology of autism with facial palsy, research into developmental abnormalities of the peripheral facial nerves is necessary. In the present study, to investigate the development of peripheral cranial nerves for use in an animal model of autism, rat embryos were treated with valproic acid (VPA) in utero and their cranial nerves were visualized by immunostaining. Treatment with VPA after embryonic day 9 had a significant effect on the peripheral fibers of several cranial nerves. Following VPA treatment, immunoreactivity within the trigeminal, facial, glossopharyngeal and vagus nerves was significantly reduced. Additionally, abnormal axonal pathways were observed in the peripheral facial nerves. Thus, the morphology of several cranial nerves, including the facial nerve, can be affected by prenatal VPA exposure as early as E13. Our findings indicate that disruption of early facial nerve development is involved in the etiology of asymmetric facial palsy, and may suggest a link to the etiology of autism.     

Immunohistochemical co-localization of lymphatics and blood vessels in oral squamous cell carcinomas

BACKGROUND: Differentiating lymphatic vessels from blood vessels is difficult, partly due to the lack of a specific method for identifying lymphatics. A new lymphatic vessel-reactive antibody, D2-40 has recently become commercially available. We examined the selectivity of D2-40 for lymphatics in oral neoplastic lesions for discrimination from blood vessels. METHODS: Formalin-fixed, paraffin-embedded sections of oral lymphangiomas (n = 3), oral hemangiomas (n = 7), and oral squamous cell carcinomas (OSCC, n = 46) were double immunostained with D2-40 and anti-CD34 monoclonal antibodies (MoAb) using ENVISION-polymer technique with 5-bromo-4-chloro-3-indoxyl-phosphate (BCIP)/nitroblue tetrazolium chloride (NBT) and 3,3'-diaminobenzidine (DAB) as color reagents, respectively. Results: In the oral lymphangiomas and hemangiomas D2-40 was detected in all lymphatics, while all blood vessels were positive for CD34. In OSCC, number of vessels for lymphatics (P < 0.01) and for blood vessels in the perineoplastic areas were significantly greater than those in intratumoral areas. CONCLUSIONS: These results indicate that lymphatic proliferation might be much more extensive in the peritumoral area than intratumoral.     

免疫组织化学和PCR在早期麻风诊断中的应用

目的：探讨PCR、免疫组化应用于早期麻风诊断的价值。方法：采用PCR、免疫组化（PGL－1）和常规病理（HE、Fite抗酸染色）3种方法并用于临床诊断麻风病，用病理检查呈轻度非特异性炎症，查菌阴性的46例石蜡组织块，重新连续切片作对比观察。结果：3种检查结果比较，PCR阳性率（58．7％）＞PGL－1（36．9％）＞抗酸染色查菌AFB（15．2％），P＜0．05。结论：PCR、免疫组化两种检测技术，用于早期少菌型麻风的诊断比常规病理检查具有较高的敏感性和特异性。     

IMMUNOELEGTRON MICROSCOPIC STUDY OF S-100 β-POSITIVE HUMAN T-LYMPHOCYTES WITH SPECIAL REFERENCE TO DOUBLE IMMUNOSTAINING

S-100 β-positive T-lymphocytes in human peripheral blood were shown to be definitely T-cells containing the S-100β subunit in their cytoplasm by double immunostaining using immunoferritin labeling for CD8 antigen and im-munoperoxidase labeling for cytoplasmic S-100 β subunit at the ultrastructural level. These results ultrastructurally confirmed the presence of double markers of S-100 β-positive T-lymphocytes seen by light microscopic double immunostaining. Furthermore, this double immunostaining method for electron microscopy is applicable to other kinds of cells for the simultaneous detection of two different antigens present both on the cell surface and in the cytoplasm. ACTA PATHOL JPN 38: 1167∼1173, 1988.