Demonstration of interleukin-3 receptor-associated antigen in the central nervous system

We previously reported that interleukin-3 (IL-3) acts as a neurotrophic factor for cholinergic neurons. However, it has not yet been determined whether the action is derived from the interaction of IL-3 with IL-3 receptors. As the first step to study IL-3 receptors in the central nervous system, we examined the presence and localization of IL-3 receptor-associated antigen (IL-3RAA) in mouse and rat brain. Immunohistochemically, IL-3RAA, which is closely involved both in the IL-3 binding to IL-3 receptors and the tyrosine phosphorylation in the signal transduction for IL-3 in hematopoietic cells, was demonstrated in neurons throughout the brain. This was confirmed in primary cultured neurons and neuronal cell lines by immunocytochemistry and flow cytometry. The staining intensity varied among regions and the most intense immunoreactivity for IL-3RAA was found in large neurons in the magnocellular basal nuclei, pyramidal cells in the cerebral cortex, and neuronal cells in some nuclei of the brainstem. Not only cholinergic cell lines derived from the septal region but also other neuronal cell lines exhibited IL-3RAA immunoreactivity by flow cytometry. Therefore, we conclude that IL-3RAA is present in a wide variety of neurons in the brain including cholinergic neurons of the basal forebrain. Western blot analysis revealed that the candidates for IL-3RAA are 145, 100, and 50 kDa proteins both in neuronal and IL-3-dependent cell lines. © 1995 Wiley-Liss, Inc.     

3H-thymidine autoradiography of CSF cells in primary reticulum cell sarcoma of the brain

Summary CSF cells in a case of primary reticulum cell sarcoma of the brain with diffuse subarachnoid spreading were examined by ³H-thymidine autoradiography. Immediately after lumbar puncture, the CSF withdrawn was incubated at 37°C for 1 hr with an admixture of ³H-thymidine at a rate of 1 Ci/ml CSF. The cells were collected by centrifugation and the microautoradiographic procedure was performed. The labeling index (L.I.) of the total CSF cells was 10.5%, and when non-neoplastic cells, i.e. polymorphonuclear leukocytes, small lymphocytes, monocytes etc., were excluded, the real L.I. of the tumor cells in the CSF was supposed to be more than 14.4%. Referring to the results of various brain tumors reported in the literature, this belongs at least to the highest labeling group. The high L. I. of the tumor cells in this case was well consistent with the extremely rapid clinical course. It should be stressed that the examination of CSF cells by ³H-thymidine autoradiography in cases of brain tumors could be one of the valuable methods indicating the DNA synthesis of the tumor cells, which is an important parameter of malignancy.

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Zusammenfassung Liquorzellen in einem Fall von primärem Reticulumzellsarkom des Gehirns mit diffuser subarachnoidealer Ausbreitung wurden mit ³H-Thymidin-Autoradiographie untersucht. Sofort nach der Lumbalpunktion wurde der entnommene Liquor bei 37°C für 1 Std mit ³H-Thymidin inkubiert. Die Zellen wurden durch Zentrifugation gesammelt, und ein mikroautoradiographisches Verfahren wurde ausgeführt. Der Markierungsindex der gesamten Liquorzellen wurde mit 10.5% errechnet. Wenn nichtneoplastische Zellen, d. h. Granulocyten, Monocyten, Lymphocyten usw., ausgeschlossen wurden, dann betrug der echte Markierungsindex der Tumorzellen im Liquor mehr als 14.4%. Der Index entspricht somit den höchsten von den in der Literatur berichteten Markierungen bösartiger Gehirntumoren des Menschen. Der hohe Markierungsindex dieses Tumors entsprach dem außerordentlich rasch progredienten klinischen Bild. Es wird hier betont, daß die Untersuchung der Liquorzellen bei Gehirntumoren mit ³H-Thymidin-Autoradiographie für eine Beurteilung der Fähigkeit der DNS-Synthese der Tumorzellen verwendet werden kann, und damit einen Hinweis auf die Malignität gibt.

     

Experimental allergic encephalomyelitis induced by proteolipid apoprotein in Lewis rats

Experimental allergic encephalomyelitis (EAE) was induced in inbred Lewis rats by sensitization with bovine white matter proteolipid apoprotein (PLP). 18-61 days after a single injection of 100 micrograms of PLP, 12 of 31 rats (39%) developed clinical EAE and 18 of 23 (78%) showed pathologic EAE with significant demyelination. Lymphocyte proliferative responses and antibodies to PLP were elevated but did not correlate with the clinical or pathologic state. This is the first demonstration of PLP-induced EAE with significant demyelination in rats and will contribute to the study of autoimmune demyelination.     

A Novel Method for Observation of the Mandibular Foramen: Application to a Better Understanding of Dental Anatomy

Cone‐beam computed tomography gives us much useful morphological information about the mandibular bone. Many studies of the mandible include findings from this technique. However, there have been no endoscopic studies of the mandible. Sixteen sides of eight dry mandibles resected from cadavers (age range 38–83 years) were examined by endoscopy. The head of the endoscope was 2.0 mm in diameter. We examined the mandibular foramen, lingula, mylohyoid groove, and mandibular canal. The mylohyoid grooves showed variations such as double grooves and canals. The mandibular lingula was located superior or medial to the the mandibular foramen. In a single case, the medial wall inside the mandibular canal showed a porous surface. The retromolar canal was observed in three sides. None of the images in the present study have been seen in other studies. Observation of the retromolar canal from the mandibular canal in particular can help dental students as well as oral and maxillofacial surgeons to understand its morphology. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 300:1875–1880, 2017. © 2017 Wiley Periodicals, Inc.     

A novel method with which to visualize the human sinuatrial node: Application for a better understanding of the gross anatomy of this part of the conduction system

For various clinical/surgical procedures, it is important to accurately understand the location of the sinuatrial node (SAN). Therefore, this study's goal was to develop a new and simple method to visualize the SAN in human hearts. A total of 16 formalin-fixed human hearts were used in the study. After the epicardium was removed, the fat tissue on the myocardium's surface was brushed and removed in a solution of 40°C water with a surfactant to show the SAN's location. Once the structure considered to be the SAN was observed, histological observation was conducted with Masson's trichrome staining to confirm its identity. The working myocardium, SAN branch of the coronary artery, and the structure believed to be the SAN were observed in all specimens. Histological analysis confirmed this structure to be the SAN. We believe that the method described herein might contribute to a better understanding of the SAN's morphologic features and serve as an improved teaching aide. Clin. Anat. 33:232–236, 2020. © 2019 Wiley Periodicals, Inc.     

Prognostic value of mitotic metaphase rate in oesophageal cancer.

OBJECTIVE: To assess the prognostic value of the mitotic metaphase rate in patients with oesophageal cancer. DESIGN: Retrospective study. SETTING: University hospital, Japan. SUBJECTS: 41 patients with oesophageal cancer. INTERVENTIONS: We calculated the ratio of mitotic metaphase to anaphase cells among tumour cells in sections stained with haematoxylin and eosin, a ratio that shows the status of mitotic metaphase-anaphase transition. The DNA ploidy pattern was examined by flow cytometry. MAIN OUTCOME MEASURE: Correlation between survival and mitotic metaphase rate. RESULTS: A high mitotic metaphase rate was correlated with vascular invasion and is expected to be a useful prognostic factor. DNA diploidy combined with a low rate was an independent favorable prognostic factor. CONCLUSION: Mitotic metaphase rate is a useful index of malignant potential, independent of DNA ploidy. It can distinguish high malignant potential from low in a diploid tumour, which has a poor prognosis that is equal to that of the aneuploidy tumour.     

Report of a patient with CADASIL having a novel missense mutation of the Notch 3 gene--association with alopecia and lumbar herniated disk]

We report a 52-year-old man with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) presenting dementia, alopecia and lumbar herniated disk. He had an episode of stroke and migraine-like headache lasting for 5 minutes. A lot of members had cerebral infarction in this family. Brain magnetic resonance imaging demonstrated, on T2-weighted images, numerous hyperintense lesions suggestive of small infarcts in the basal ganglia and diffuse hyperintense lesions in the cerebral white matter. The clinical symptoms, the family history and the MRI findings suggested the diagnosis of CADASIL. However, the patient also showed alopecia and lumbar herniated disk, both are characteristic features of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). The DNA analysis of the Notch 3 gene identified a novel missense mutation Cys174Phe in this patient. Our case report indicated the importance of the DNA analysis for the diagnosis of CADASIL.     

A novel monoclonal antibody specific for the amino-truncated beta-amyloid Abeta5-40/42 produced from caspase-cleaved amyloid precursor protein

Deposition of beta-amyloid peptide (Abeta) as senile plaques and amyloid angiopathy are the major neuropathological features of Alzheimer's disease (AD). Heterogeneity is observed in the N- and C-termini of the deposited Abeta species. Recent evidence implicates caspase activation and apoptosis in AD neurodegeneration. We previously reported that a distinct N-terminally truncated Abeta species, Abeta5-40/42 is preferentially produced from the caspase-cleaved form of amyloid precursor protein (APP) lacking its C-terminal 31 amino acids and that it is deposited in AD brain tissues. Here, we generated a novel monoclonal antibody specific to the N-terminal end of Abeta5-40/42. Western blotting confirmed that this antibody recognizes Abeta5-40 but not Abeta1-40. We also showed that the antibody is able to immunoprecipitate Abeta5-40 but not Abeta1-40. Immunoprecipitation with the antibody followed by mass spectrometric analysis further detected Abeta5-40 in the conditioned media from neuroblastoma cells expressing the caspase-cleaved APP. The antibody reacted weakly with Abeta derived from AD brains. These results suggest that our novel monoclonal antibody is useful for detecting the N-terminally truncated Abeta produced in conjunction with caspase activation.     

Amyloid beta protein-related death-inducing protein induces G2/M arrest: Implications for neurodegeneration in Alzheimer's disease

Amyloid beta protein (Abeta)-related death-inducing protein (AB-DIP) is a novel Abeta binding protein expressed ubiquitously. Here we demonstrate that overexpression of AB-DIP in SH-SY5Y neuroblastoma cells causes G2/M arrest. By deletion mutant analysis, we have identified the minimal region within AB-DIP required for G2/M arrest. We have also shown that microtubule-interfering agents (MIAs) such as nocodazole, vinblastine, paclitaxel, and vincristine, known to arrest cells at G2/M, also phosphorylate AB-DIP. However, etoposide, which causes genotoxic stress; tunicamycin, an ER stress inducer; and rotenone, which causes mitochondrial damage, fail to phosphorylate AB-DIP, implying that phosphorylation of AB-DIP is specific to microtubule-disruption-induced G2/M arrest. By using different classes of kinase inhibitors, we also demonstrate that a putative tyrosine kinase phosphorylates AB-DIP. Mono- or multisite mutations of tyrosine or serine/threonine residues confirmed that mutation of tyrosine residues but not serine/threonine residues greatly reduces nocodazole-induced phosphorylation of AB-DIP. Furthermore, phosphorylation of AB-DIP can be induced in MCF-7 cells that lack functional p53, suggesting that AB-DIP phosphorylation is independent of p53. Mounting experimental evidence continues to support the role of cell cycle abnormalities in the pathogenesis of Alzheimer's disease, and our results suggest that AB-DIP might provide a mechanistic link between microtubule disruption, mitotic abnormalities, neuronal dysfunction, and death. Therefore, interfering with AB-DIP may have therapeutic applications in conditions such as Alzheimer's disease, in which microtubule disruption and mitotic abnormalities have been suggested to play a pathological role.