Electron microscopic and functional studies on platelets in gray platelet syndrome

The ultrastructure and functions of platelets and megakaryocytes in 8 patients of a family with gray platelet syndrome were investigated. Hemostatic examinations on these patients revealed prolonged bleeding time, decreased platelet retention rates and decreased platelet aggregation rates induced by ADP, collagen, Ristocetin and epinephrine. Marked decreases in ATP and ADP release in response to these agents were also noted. Clinical and coagulation studies on this family suggested that the hereditary nature of the syndrome is autosomal dominant. Platelets and megakaryocytes in the peripheral and bone marrow blood smear from the patients showed peculiar gray color by May-Giemsa stain due to a deficiency of alpha-granules. Electron microscopic examinations revealed slightly enlarged platelets containing a deficient amount of alpha-granules, whereas dense bodies and mitochondria appeared normal. Several morphological abnormalities of patient's platelets, such as aggregates of dense tubular systems, circular arrays of dense tubular systems, an area of cytoplasmic sequestration with an enclosing membrane, clumps of dense material and remnants of Golgi apparatuses were recognized. Megakaryocytes showed normally developed Golgi zones, defective alpha-granule synthesis and liberation of abnormal platelets as shown in the peripheral blood smear.     

Changes in fetal plasma adenosine and xanthine concentrations during fetal asphyxia with maternal oxygen administration in ewes

In this study, we measured fetal plasma adenosine and xanthine concentrations during and after severe asphyxia, and investigated the key issues related to oxygen therapy. Asphyxia was induced by occluding the umbilical cord for 5 minutes in 6 fetal sheep with and without the administration of oxygen to the ewe. Plasma adenosine concentration increased significantly during cord occlusion in the all fetuses, and the differences between the values in the fetuses with and without maternal oxygen administration was not significant. By 30 minutes after cord release, plasma adenosine concentration in all fetuses had returned to levels similar to those at the start of the experiment. Plasma xanthine concentration also increased during cord occlusion in all fetuses. However, 30 minutes after cord release, plasma xanthine concentration had decreased significantly in fetuses without additional oxygen, while it did not change significantly in fetuses with maternal oxygen administration. Thus, we speculated that maternal oxygen administration before fetal asphyxia may not contribute to additional ATP stores in fetal organs and may produce oxygen free radicals following asphyxia.     

Morphologic study of cytoskeletal systems of mouse hepatocytes using polyethylene glycol-embedding method

SM mouse livers extracted by immersion in 1% Triton X-100, or in 1% Triton X-100 followed by 0.3 M KI were studied electron microscopically using the polyethylene glycol-embedding method. After extraction with 1% Triton X-100, almost all the structural components of hepatocytes remained intact and cytoplasmic filaments could be seen three-dimensionally by using stereopairs of micrographs. It was difficult, however, to discriminate microfilaments, intermediate-sized filaments and microtubules from one anoter in these specimes . By immersion in 1% Triton X-100 followed by 0.3 M KI, hepatocytes were extracted remaining only plasma membranes, vesicles and filaments. These filaments were approximately 10 nm in diameter, that is intermediate in size. They were branched and were connected with plasma membranes, especilly at desmosomes. The combination method of immersion extraction and PEG-embedding seems to be suitable for the electron microscopic observation of the cytoskeleton of cells in situ.     

“Gliomatosis encephali” as a novel category of brain tumors by the first autopsy case report of gliomatosis cerebelli

Gliomatosis cerebri is a rare diffuse glioma that is neither mass‐forming nor necrotic, and does not disrupt existing structures. Gliomatosis occurring in the cerebellum is known as gliomatosis cerebelli, and only three such cases examined by biopsy have been reported. Here we describe the first autopsy findings of a patient who was diagnosed as having gliomatosis in the cerebellum. Neuropathological examination identified the tumor cells as being positive for glial fibrillary acidic protein, vimentin and nestin, with atypical nuclei that were cashew‐nut‐ or dishcloth‐gourd‐shaped. These tumor cells were dense in the right cerebellum, but also spread broadly throughout the brain including the left cerebrum and optic nerve. Mitotic figures were frequently seen in the cerebellum, brain stem and cerebrum. Scherer's secondary structures were evident not only in the cerebellum but also the cerebrum. No necrosis, microvascular proliferation or destruction of anatomical structures was detected in the whole brain. Differences in the origin of the tumors of the gliomatoses cerbri and cerebelli suggests these tumors are different types of brain tumors. Thus the findings support that the gliomatosis cerebelli is a novel type of brain tumor classification. Furthermore, by the similarities of the histological features among the tumors, it appears appropriate to establish a novel category of “gliomatosis encephali” which includes both gliomatosis cerebri and gliomatosis cerebelli.     

Regulatory Mechanisms of RANKL Presentation to Osteoclast Precursors

It is important to understand the molecular mechanisms regulating osteoclast formation, as excess activation of osteoclasts is associated with various osteopenic disorders. Receptor activator of nuclear factor kappa B (RANKL) is a central player in osteoclastogenesis. Recent findings suggest that osteocytes are the major supplier of RANKL to osteoclast precursors. It has also been suggested that osteocyte cell death upregulates the RANKL/osteoprotegerin (OPG) ratio in viable osteocytes adjacent to apoptotic osteocytes in areas of bone microdamage, thus, contributing to localized osteoclast formation. Indeed, viable osteocytes can provide RANKL through direct interactions with osteoclast precursors at osteocyte dendritic processes. In addition, OPG tightly regulates RANKL cell surface presentation in osteocytes, which contributes to the inhibition of RANKL signaling, as well as the decoy receptor function of OPG. By contrast, the physiological role of RANKL in osteoblasts is yet to be clarified, although similar mechanisms of regulation are observed in both osteocytes and osteoblasts.     

Common variants at 1p36 are associated with superior frontal gyrus volume

The superior frontal gyrus (SFG), an area of the brain frequently found to have reduced gray matter in patients with schizophrenia, is involved in self-awareness and emotion, which are impaired in schizophrenia. However, no genome-wide association studies of SFG volume have investigated in patients with schizophrenia. To identify single-nucleotide polymorphisms (SNPs) associated with SFG volumes, we demonstrated a genome-wide association study (GWAS) of gray matter volumes in the right or left SFG of 158 patients with schizophrenia and 378 healthy subjects. We attempted to bioinformatically ascertain the potential effects of the top hit polymorphism on the expression levels of genes at the genome-wide region. We found associations between five variants on 1p36.12 and the right SFG volume at a widely used benchmark for genome-wide significance (P<5.0 × 10⁻⁸). The strongest association was observed at rs4654899, an intronic SNP in the eukaryotic translation initiation factor 4 gamma, 3 (EIF4G3) gene on 1p36.12 (P=7.5 × 10⁻⁹). No SNP with genome-wide significance was found in the volume of the left SFG (P>5.0 × 10⁻⁸); however, the rs4654899 polymorphism was identified as the locus with the second strongest association with the volume of the left SFG (P=1.5 × 10⁻⁶). In silico analyses revealed a proxy SNP of rs4654899 had effect on gene expression of two genes, HP1BP3 lying 3′ to EIF4G3 (P=7.8 × 10⁻⁶) and CAPN14 at 2p (P=6.3 × 10⁻⁶), which are expressed in moderate-to-high levels throughout the adult human SFG. These results contribute to understand genetic architecture of a brain structure possibly linked to the pathophysiology of schizophrenia.