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High expression of gangliosides GD3 and GD2 is observed in human gliomas. The functions of GD3 and GD2 in malignant properties have been reported in glioma cells in vitro, but those functions have not yet been investigated in vivo. In this study, we showed that deficiency of GD3 synthase (GD3S, St8sia1) attenuated glioma progression and clinical and pathological features in a platelet‐derived growth factor B‐driven murine glioma model. Lack of GD3S resulted in the prolonged lifespan of glioma‐bearing mice and low‐grade pathology in generated gliomas. Correspondingly, they showed reduced phosphorylation levels of Akt, Erks, and Src family kinases in glioma tissues. A DNA microarray study revealed marked alteration in the expression of various genes, particularly in MMP family genes, in GD3S‐deficient gliomas. Re‐expression of GD3S restored expression of MMP9 in primary‐cultured glioma cells. We also identified a transcription factor, Ap2α, expressed in parallel with GD3S expression, and showed that Ap2α was critical for the induction of MMP9 by transfection of its cDNA and luciferase reporter genes, and a ChIP assay. These findings suggest that GD3S enhances the progression of gliomas by enhancement of the Ap2α‐MMP9 axis. This is the first report to describe the tumor‐enhancing functions of GD3S in vivo., Genetically engineered mice were used for evaluation of ganglioside GD3 synthase in murine gliomas. Deficiency of GD3 synthase resulted in prolonged life‐span of glioma‐bearing mice and low grade pathology in generated gliomas. Reduced activation of signaling molecules and MMP proteins in the KO mice has been described.

Loading is indispensable for the growth, development, and maintenance of joint tissues, including mandibular condylar cartilage, but excessive loading or reduced host adaptive capacity can considerably damage the temporomandibular joint (TMJ), leading to temporomandibular joint osteoarthritis (TMJ-OA). TMJ-OA, associated with other pathological conditions and aging processes, is a highly degenerative disease affecting the articular cartilage. Many treatment modalities for TMJ-OA have been developed. Traditional clinical treatment includes mainly nonsurgical options, such as occlusal splints. However, non-invasive therapy does not achieve joint tissue repair and regeneration. Growing evidence suggests that low-intensity pulsed ultrasound (LIPUS) accelerates bone fracture healing and regeneration, as well as having extraordinary effects in terms of soft tissue repair and regeneration. The latter have received much attention, and various studies have been performed to evaluate the potential role of LIPUS in tissue regeneration including that applied to articular cartilage. The present article provides an overview of the status of LIPUS stimulation used to prevent the onset and progression of TMJ-OA and enhance the tissue regeneration of mandibular condylar cartilage. The etiology and management of TMJ-OA are explained briefly, animal models of TMJ-OA are described, and the effectiveness of LIPUS on cell metabolism and tissue regeneration in the TMJ is discussed.

Summary Objective Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease characterized by progressive cartilage degeneration, abnormal bone remodeling, and chronic pain. In this study, we aimed to investigate effective therapies to reverse or suppress TMJOA progression. Design To this end, we performed intravenous administration of serum free conditioned media from human exfoliated deciduous teeth stem cells (SHED-CM) into a mechanical-stress induced murine TMJOA model. Results SHED-CM administration markedly suppressed temporal muscle inflammation, and improved bone integrity and surface smoothness of the destroyed condylar cartilage. Moreover, SHED-CM treatment decreased the number of IL-1β, iNOS, and MMP-13 expressing chondrocytes, whereas it specifically increased PCNA-positive cells in the multipotent polymorphic cell layer. Notably, the numbers of TdT-mediated dUTP nick end labeling (TUNEL)-positive apoptotic chondrocytes in the SHED-CM treated condyles were significantly lower than in those treated with DMEM, whereas the proteoglycan positive area was restored to a level similar to that of the sham treated group, demonstrating that SHED-CM treatment regenerated the mechanical-stress injured condylar cartilage and subchondral bone. Secretome analysis revealed that SHED-CM contained multiple therapeutic factors that act in osteochondral regeneration. Conclusions Our data demonstrated that SHED-CM treatment promoted the regeneration and repair of mechanical-stress induced mouse TMJOA. Our observations suggest that SHED-CM has potential to be a potent tissue-regenerating therapeutic agent for patients with severe TMJOA.

To analyze the binding specificity of a sialic acid–recognizing lectin, sialic acid-binding Ig-like lectin 7 (SIGLEC7), to disialyl gangliosides (GD3s), here we established GD3-expressing cells by introducing GD3 synthase (GD3S or ST8SIA1) cDNA into a colon cancer cell line, DLD-1, that expresses no ligands for the recombinant protein SIGLEC7-Fc. SIGLEC7-Fc did not recognize newly-expressed GD3 on DLD-1 cells, even though GD3 was highly expressed, as detected by an anti-GD3 antibody. Because milk-derived GD3 could be recognized by this fusion protein when incorporated onto the surface of DLD-1 cells, we compared the ceramides in DLD-1–generated and milk-derived GD3s to identify the SIGLEC7-specific GD3 structures on the cell membrane, revealing that SIGLEC7 recognizes only GD3-containing regular ceramides but not phytoceramides. This was confirmed by knockdown/knockout of the sphingolipid delta(4)-desaturase/C4-monooxygenase (DES2) gene, involved in phytoceramide synthesis, disclosing that DES2 inhibition confers SIGLEC7 binding. Furthermore, knocking out fatty acid 2-hydroxylase also resulted in the emergence of SIGLEC7 binding to the cell surface. To analyze the effects of binding between SIGLEC7 and various GD3 species on natural killer function, we investigated cytotoxicity of peripheral blood mononuclear cells from healthy donors toward GD3S-transfected DLD-1 (DLD-1–GD3S) cells and DLD-1–GD3S cells with modified ceramides. We found that cytotoxicity is suppressed in DLD-1–GD3S cells with dehydroxylated GD3s. These results indicate that the ceramide structures in glycosphingolipids affect SIGLEC7 binding and distribution on the cell surface and influence cell sensitivity to killing by SIGLEC7-expressing effector cells.

Cancer-associated glycosphingolipids have been used as markers for diagnosis and targets for immunotherapy of malignant tumors. Recent progress in the analysis of their implications in the malignant properties of cancer cells revealed that cancer-associated glycosphingolipids are not only tumor markers, but also functional molecules regulating various signals introduced by membrane microdomains, lipid rafts. In particular, a novel approach, enzyme-mediated activation of radical sources combined with mass spectrometry, has enabled us to clarify the mechanisms by which cancer-associated glycosphingolipids regulate cell signals based on the interaction with membrane molecules and formation of molecular complexes on the cell surface. Novel findings obtained from these approaches are now providing us with insights into the development of new anticancer therapies targeting membrane molecular complexes consisting of cancer-associated glycolipids and their associated membrane molecules. Thus, a new era of cancer-associated glycosphingolipids has now begun.

BackgroundAlthough recent studies have revealed the powerful antinociceptive effect of human dental pulp stem cells in an animal model for diabetes and osteoarthritis, its analgesic mechanisms are still largely elusive. We have previously reported that conditioned medium (CM) from dental pulp stem cells of deciduous teeth (SHED-CM) or its components, monocyte chemoattractant protein-1 (MCP-1) and the secreted ectodomain of sialic acid-binding Ig-like lectin-9 (sSiglec-9), directly induces anti-inflammatory M2 macrophages, however the antinociceptive activity of induced M2 is unknown. In this study, we investigated the antinociceptive effect of SHED-CM, MCP-1, and sSiglec-9 or secretome from M2-induced by SHED-CM (M2-CM) against neuropathic pain (NP) using a partial sciatic nerve ligation (PSL) mouse model and analyzed the mechanical bases of their antinociceptive effects. MethodsPSL mice were treated using SHED-CM with or without mannosylated-Clodrosome, specifically depleting M2 macrophages, recombinant MCP-1 and sSiglec-9 protein, M2-CM, or fibroblast-CM. Human Schwann cells activated by TNF-α in vitro were treated with M2-CM. The expression of pro-inflammatory mediators, neuroprotective factors, the nociceptive receptor, and markers for M1, M2, and activated glial cells in injured sciatic nerve (SCN), dorsal root ganglion, or spinal cord were evaluated by RT-PCR and immunohistochemistry. Mechanical allodynia of PSL mice was analyzed via von Frey test. ResultsIn the behavioral test, intravenous administration of SHED-CM greatly improved the PSL-induced hypersensitivity. SHED-CM treatment recruited M2 macrophages in the injured SCN and ipsilateral L4/L5 dorsal root ganglion and suppressed microglial activation in the spinal cord. Specific depletion of the M2 by mannosylated-Clodrosome markedly reduced the antinociceptive effect of SHED-CM. Intravenous administration of both MCP-1/sSiglec-9 and M2-CM ameliorated the PSL-induced hypersensitivity. We found that M2-CM directly suppressed the expression of nociceptive receptors as well as proinflammatory mediators in Schwann cells. ConclusionTaken together, our data suggest that SHED-CM ameliorates NP through the induction of the analgesic anti-inflammatory M2 macrophages. Thus, SHED-CM may present as a potential novel therapeutic candidate for NP.

Ganglioside GD2 is specifically expressed in small-cell lung cancer (SCLC) cells, leading to enhancement of malignant phenotypes, such as cell proliferation and migration. However, how GD2 promotes malignant phenotypes in SCLC cells is not well known. In this study, to reveal the mechanisms by which GD2 increases malignant phenotypes in SCLC cells, we used enzyme-mediated activation of radical sources combined with mass spectrometry in GD2+ SCLC cells. Consequently, we identified ASC amino acid transporter 2 (ASCT2), a major glutamine transporter, which coordinately works with GD2. We showed that ASCT2 was highly expressed in glycolipid-enriched microdomain/rafts in GD2+ SCLC cells, and colocalized with GD2 in both proximity ligation assay and immunocytostaining, and bound with GD2 in immunoprecipitation/TLC immunostaining. Malignant phenotypes of GD2+ SCLC cells were enhanced by glutamine uptake, and were suppressed by L-γ-glutamyl-p-nitroanilide, a specific inhibitor of ASCT2, through reduced phosphorylation of p70 S6K1 and S6. These results suggested that ASCT2 enhances glutamine uptake in glycolipid-enriched microdomain/rafts in GD2+ SCLC cells, leading to the enhancement of cell proliferation and migration through increased phosphorylation of the mTOR complex 1 signaling axis.

Gene knockout mice of glycosyltransferases have clearly showed roles of their products in the bodies, while there are examples where phenotype of knockout was much less severe than expected probably due to functional redundancy. The most striking novel finding obtained from ganglioside-deficient mice was that progressive inflammatory reaction took place, leading to neurodegeneration. In particular, dysfunction of complement-regulatory proteins due to deteriorated architecture of lipid rafts seemed to be essential mechanisms for the inflammation. Furthermore, roles of gangliosides in neurons were demonstrated by neuron-specific transgenic of B4galnt1 with genetic background of B4galnt1 deficiency. From study of gene knockout mice of St8sia1, new roles of b-series gangliosides in leptin secretion from adipocytes, and roles of a-series gangliosides in leptin receptor, ObR in hypothalamus were demonstrated, leading to apparent intact balance of energy. Essential roles of b-series gangliosides in malignant properties of gliomas were also shown, suggesting their roles in the regulation of inflammation and proliferation in nervous tissues. How to apply these findings for the control of newly discovered patients with ganglioside deficiency remains to be investigated. This article is part of a Special Issue entitled Neuro-glycoscience, edited by Kenji Kadomatsu and Hiroshi Kitagawa.

Acidic glycosphingolipids, i.e., gangliosides, are predominantly and consistently expressed in nervous tissues of vertebrates at high levels. Therefore, they are considered to be involved in the development and function of nervous systems. Recent studies involving genetic engineering of glycosyltransferase genes have revealed novel aspects of the roles of gangliosides in the regulation of nervous tissues. In this review, novel findings regarding ganglioside functions and their modes of action elucidated mainly by studies of gene knockout mice are summarized. In particular, the roles of gangliosides in the regulation of lipid rafts to maintain the integrity of nervous systems are reported with a focus on the roles in the regulation of neuro-inflammation and neurodegeneration via complement systems. In addition, recent advances in studies of congenital neurological disorders due to genetic mutations of ganglioside synthase genes and also in the techniques for the analysis of ganglioside functions are introduced.

Some gangliosides, sialic acid-containing glycosphingolipids, have been considered as tumor-associated antigens. GD1α or a GD1α synthase gene ST6GalNAc5 was reported to be involved in the metastasis of murine lymphomas or human breast cancers, respectively. But expression patterns of 0-series gangliosides GD1α and its precursor GM1b in human cancers have not yet been investigated mainly due to lack of specific antibodies. We established specific monoclonal antibodies (mAbs) reactive with GD1α or GM1b using gangliosides from brain tissues of GM3 synthase (St3gal5)-deficient mice as immunogens. We used GM2/GD2 synthase (B4galnt1)-deficient mice to immunize by liposomes embedded with GD1α or acidic glycolipid fractions from brain of St3gal5-deficient mice. Specificities of established mAbs as analyzed by enzyme-linked immunosorbent assay and thin-layer chromatography-immunostaining were very high among various gangliosides. Increased expression of GD1α and reduced GM1b in the St6galnac5 cDNA-transfected RAW117 cell line also substantiated the specificities of two mAbs. Then, we analyzed expression of GD1α and GM1b, and of relevant glycosyltransferase genes in various human cancer cell lines using generated anti-GD1α mAb 122 or anti-GM1b mAb MR155A-7. A few human cancer cell lines showed significant expression of these gangliosides with reasonable expression of relevant glycosyltransferase genes.

B4GALNT1 is an enzyme essential for the synthesis of complex gangliosides, whose absence leads to progressive neurodegeneration with aging in mice. Recently, eleven cases of hereditary spastic paraplegia with mutation in the coding region of B4GALNT1 were reported. However, changes in the enzymatic activity of their products have never been studied. We have constructed expression vectors for individual mutant cDNAs, and examined their activities by cell-free in vitro enzyme assays, and flow cytometry of cells transfected with their expression vectors. Among them, almost all mutant genes showed the complete loss of B4GALNT1 activity in both the in vitro enzyme assays and flow cytometry. Two mutants exceptionally showed weak activity. One of them, M4, had a mutation at amino acid 228 with a premature termination codon. Interestingly, the intensity of fluorescence of GM2 measured by flow cytometry was equivalent between the WT and M4 mutant, although the positive cell population was relatively small in M4. Western immunoblotting of cell lysates from transfectants with cDNA plasmids revealed 67-kDa bands except those containing premature termination codons or frame-shift mutation. Taken together with the clinical findings of patients, loss of enzyme activity may be responsible for the clinical features of hereditary spastic paraplegia, whereas the intensity of neurological disorders was relatively milder than expected. These clinical features of patients including those with male hypogonadism are very similar to the abnormal phenotypes detected in B4galnt1-deficient mice.

In this study, we immunized Gb3/CD77 synthase gene (A4galt) knockout (KO) mice with glycosphingolipids (GSLs) extracted from 3 renal cell cancer (RCC) cell lines to raise monoclonal antibodies (mAbs) reactive with globo-series GSLs specifically expressed in RCCs. Although a number of mAbs reactive with globo-series GSLs were generated, they reacted with both RCC cell lines and normal kidney cells. When we analyzed recognized antigens by mAbs that were specifically reactive with RCC, but not with normal kidney cells at least on the cell surface, many of them turned out to be reactive with sulfoglycolipids. Eight out of 11 RCC-specific mAbs were reactive with SM2 alone, and the other 3 mAbs were more broadly reactive with sulfated glycolipids, i.e. SM3 and SM4 as well as SM2. In the immunohistochemistry, these anti-sulfoglycolipids mAbs showed RCC-specific reaction, with no or minimal reaction with adjacent normal tissues. Thus, immunization of A4galt KO mice with RCC-derived GSLs resulted in the generation of anti sulfated GSL mAbs, and these mAbs may be applicable for the therapeutics for RCC patients.

There have been a few studies on the ganglioside expression in human glioma tissues. However, the role of these gangliosides such as GD3 and GD2 has not been well understood. In this study we employed a genetically engineered mouse model of glioma to clarify the functions of GD3 in gliomas. Forced expression of platelet-derived growth factor B in cultured astrocytes derived from p53-deficient mice resulted in the expression of GD3 and GD2. GD3-positive astrocytes exhibited increased cell growth and invasion activities along with elevated phosphorylation of Akt and Yes kinase. By enzyme-mediated activation of radical sources reaction and mass spectrometry, we identified PDGF receptor α (PDGFRα) as a GD3-associated molecule. GD3-positive astrocytes showed a significant amount of PDGFRα in glycolipid-enriched microdomains/rafts compared with GD3-negative cells. Src kinase family Yes was co-precipitated with PDGFRα, and its pivotal role in the increased cell invasion of GD3-positive astrocytes was demonstrated by silencing with anti-Yes siRNA. Direct association between PDGFRα and GD3 was also shown, suggesting that GD3 forms ternary complex with PDGFRα and Yes. The fact that GD3, PDGFRα, and activated Yes were colocalized in lamellipodia and the edge of tumors in cultured cells and glioma tissues, respectively, suggests that GD3 induced by platelet-derived growth factor B enhances PDGF signals in glycolipid-enriched microdomain/rafts, leading to the promotion of malignant phenotypes such as cell proliferation and invasion in gliomas.

Gangliosides play roles in the regulation of cell signaling that are mediated via membrane microdomains, lipid rafts. In this review, functions of gangliosides in the maintenance of nervous systems with a focus on regulation of inflammation and neurodegeneration are addressed. During analyses of various ganglioside-lacking mutant mice, we demonstrated that nervous tissues exhibited inflammatory reactions and subsequent neurodegeneration. Among inflammation-related genes, factors of the complement system showed up-regulation with aging. Analyses of architectures and compositions of lipid rafts in nervous tissues from these mutant mice revealed that dysfunctions of complement regulatory proteins based on disrupted lipid rafts were main factors to induce the inflammatory reactions resulting in neurodegeneration. Ganglioside changes in development and senescence, and implication of them in the integrity of cell membranes and cellular phenotypes in physiological and pathological conditions including Alzheimer disease have been summarized. Novel directions to further analyze mechanisms for ganglioside functions in membrane microdomains have been also addressed.

Glycosphingolipids are expressed on the cell membrane and act as important factors in various events that occur across the plasma membrane. Lactosylceramide (LacCer) is synthesized from glucosylceramide and is a common precursor of various glycosphingolipids existing in whole body. Based on the enzyme purification, β1,4-galactosyltransferase 6 (B4galt6) cDNA was isolated as a LacCer synthase-coding gene in the rat brain. We generated B4galt6 gene knockout (KO) mice and analyzed their phenotypes to examine roles of β4GalT6. B4galt6 KO mice were born and grew up apparently normal. LacCer synthase activity and the composition of acidic glycosphingolipids in the brain were almost equivalent or minimally different between wild-type and KO mice. Studies by mouse embryonic fibroblasts (MEFs) revealed that the silencing of B4galt5 gene resulted in the marked reduction in LacCer synthase activity and this reduction was more severe in MEFs derived from B4galt6 KO mice than those from wild-type mice. These results suggested that β4GalT6 plays a role as a LacCer synthase, whereas β4GalT5 acts as a main enzyme for LacCer biosynthesis in these tissues and cells.

Using mouse and human monoclonal antibodies, a number of cancer-associated antigens were defined, and many of their epitopes have turned out to be carbohydrates. Besides as a tumor marker, some of them really play roles as regulatory molecules on the cell membrane. Generally, disialyl glycosphingolipids (disialyl gangliosides) such as GD3 and GD2 transduce activating signals to enhance malignant properties of cancer cells, while monosialyl gangliosides such as GM1 transduce suppressive signals on cancer properties. In particular, roles of GD3 in melanomas and GD2 in small cell lung cancers, osteosarcomas, and breast cancers have been well investigated. Recently, roles of GM1 in the suppression of cancer metastasis have been demonstrated using a murine lung cancer system. These regulatory functions of cancer-associated glycolipids are performed at membrane microdomains named glycolipid-enriched microdomain (GEM)/rafts. As mechanisms for these regulatory functions, molecular interactions between glycolipids and various membrane proteins existing in the vicinity of them at GEM/rafts are of importance and now under vigorous investigation.

To investigate mechanisms for increased malignant properties in malignant melanomas by ganglioside GD3, enzyme-mediated activation of radical sources and subsequent mass spectrometry were performed using an anti-GD3 antibody and GD3-positive (GD3+) and GD3-negative (GD3-) melanoma cell lines. Neogenin, defined as a GD3-neighbored molecule, was largely localized in lipid/rafts in GD3+ cells. Silencing of neogenin resulted in the reduction of cell growth and invasion activity. Physical association between GD3 and neogenin was demonstrated by immunoblotting of the immunoprecipitates with anti-neogenin antibody from GD3+ cell lysates. The intracytoplasmic domain of neogenin (Ne-ICD) was detected in GD3+ cells at higher levels than in GD3- cells when cells were treated by a proteasome inhibitor but not when simultaneously treated with a γ-secretase inhibitor. Exogenous GD3 also induced increased Ne-ICD in GD3- cells. Overexpression of Ne-ICD in GD3- cells resulted in the increased cell growth and invasion activity, suggesting that Ne-ICD plays a role as a transcriptional factor to drive malignant properties of melanomas after cleavage with γ-secretase. γ-Secretase was found in lipid/rafts in GD3+ cells. Accordingly, immunocyto-staining revealed that GD3, neogenin, and γ-secretase were co-localized at the leading edge of GD3+ cells. All these results suggested that GD3 recruits γ-secretase to lipid/rafts, allowing efficient cleavage of neogenin. ChIP-sequencing was performed to identify candidates of target genes of Ne-ICD. Some of them actually showed increased expression after expression of Ne-ICD, probably exerting malignant phenotypes of melanomas under GD3 expression.

Engrafted mesenchymal stem cells from human deciduous dental pulp (SHEDs) support recovery from neural insults via paracrine mechanisms that are poorly understood. Here we show that the conditioned serum-free medium (CM) from SHEDs, administered intrathecally into rat injured spinal cord during the acute postinjury period, caused remarkable functional recovery. The ability of SHED-CM to induce recovery was associated with an immunoregulatory activity that induced anti-inflammatory M2-like macrophages. Secretome analysis of the SHED-CM revealed a previously unrecognized set of inducers for anti-inflammatory M2-like macrophages: monocyte chemoattractant protein-1 (MCP-1) and the secreted ectodomain of sialic acid-binding Ig-like lectin-9 (ED-Siglec-9). Depleting MCP-1 and ED-Siglec-9 from the SHED-CM prominently reduced its ability to induce M2-like macrophages and to promote functional recovery after spinal cord injury (SCI). The combination of MCP-1 and ED-Siglec-9 synergistically promoted the M2-like differentiation of bone marrow-derived macrophagesin vitro, and this effect was abolished by a selective antagonist for CC chemokine receptor 2 (CCR2) or by the genetic knock-out of CCR2. Furthermore, MCP-1 and ED-Siglec-9 administration into the injured spinal cord induced M2-like macrophages and led to a marked recovery of hindlimb locomotor function after SCI. The inhibition of this M2 induction through the inactivation of CCR2 function abolished the therapeutic effects of both SHED-CM and MCP-1/ED-Siglec-9. Macrophages activated by MCP-1 and ED-Siglec-9 extended neurite and suppressed apoptosis of primary cerebellar granule neurons against the neurotoxic effects of chondroitin sulfate proteoglycans. Our data suggest that the unique combination of MCP-1 and ED-Siglec-9 repairs the SCI through anti-inflammatory M2-like macrophage induction.

Gamma nailing is in principle performed for fractures proximal to the femur at our hospital. We conducted intraoperative and postoperative investigations in 97 cases who had undergone gamma nailing, and studied 86 cases for which follow-up data were available. This paper is a report of our findings.Complications were found in 2.3% intraoperatively and in 5.8% postoperatively.Studies of indications for surgery with consideration given to the medullary space diameter, complete reduction of the fractured region, skill in surgical techniques and regular postoperative follow-up are considered important as measures to deal with complications.

Of 213 patients who were treated for femoral neck fracture at our hospital 153 patients could be followed up. We investigated their condition and walking ability after discharge, and studied the ideal liaison medical service for this fracture. Twenty percent of these patients were sent home after discharg while the remaining 80 percent changed to another hospital. This shows the importance of liaison medical service, including welfare, for this fracture. Our department has prepared a critical path and is using it to dissipate anxiety and dissatisfaction of patients' families over the liaison medical service, and also to maintain the quality of medical care at hospitals receiving these patients. We plan to prepare a critical path incorporating the opinions of liaison hospitals for the overall improvement of the liaison medical service. In this paper, we report on our methods for carrying out the liaison medical service efficiently.

Orthognathic surgery was performed on 362 patients from 1987 to 1997 at the Second Department of Oral and Maxillofacial Surgery, Osaka Dental University.The results were as follows: 1. The male-female ratio was 1: 2.4.2. The mean age was 22.4 years old.3. The diagnosis of mandibular prognathism was made for most of the patients (89.2%).4. Sagittal splitting ramus osteotomy (SSRO) was performed on 274 cases (75.7%).5. The mean time of operation for SSRO was 3hr. 7 min±57min, and for mandibular body osteotomy was 3hr. 15min±60.7min.6. The mean blood loss during operation for SSRO was 275.8±221.4ml and for mandibular body osteotomy was 171.8±147.5ml.7. Paresthesia of the lip and mental area was observed in 404 sides after opration. 80.7% of patients treated for SSRO recovered within a year.8. TMJ were examined in 202 cases for 1 year after SSRO. The symptom of TMJ was recognized in 5 cases.

The activity indices of the masticatory and temporal muscles and the maxillofacial morphologies in patients with skeletal mandibular prognathism at the initial examination were compared with those in individuals with normal occlusion. The results were as follows:1. The muscle activity index was significantly lower in the prognathic patients than in the normal individuals.2. The items of maxillofacial morphometry that correlated with the muscle activity index were fewer and weaker in the prognathic patients than in the normal individuals.These findings suggested that patients with skeletal mandibular prognathism have functional and morphological disturbances.

The purpose of this study was to clarify the effect of long-term exercise on the maintenance of concentration, as an indicator of the activity level of the cerebral cortex, and on the Profile of Mood States (POMS) in healthy elderly persons. Forty-six subjects were divided into three groups: a long-term exercise group (LTE-G) of 21 subjects (mean age: 67 years); a health exercise group (HE-G) of 12 subjects (mean age: 69 years); and a sedentary control group (SC-G) of 13 subjects (mean age: 70 years). We assessed the target aiming function (TAF), the POMS, and the presence in the urine of the following compounds: 8-hydroxydeoxyguanosine (8-OHdG) and acrolein (ACR) as indicators of oxidative stress; adrenaline, noradrenaline, and dopamine as indicators of sympathetic nervous activity. The mean TAF-L values, which represent the level of mental concentration, were statistically lower in the LTE-G and HE-G than in the SC-G. The mean TAF-D values, indicating deviation levels, were significantly lower in the HE-G than in the SC-G. Total Mood Disturbance (TMD) scores, which are the scored totals of POMS tests, and the scores on the "Confusion" factor were clearly lower in the LTE-G and HE-G than in the SC-G. On the other hand, there were no significant differences in 8-OHdG, ACR, adrenaline, noradrenaline or dopamine values among the three groups. These results suggest that the continuation of exercise improves the cerebral cortical activity, the maintenance of concentration, and the POMS scores of healthy elderly persons.

Determining the oxygenation level of skeletal muscle is an effective non-invasive method of evaluating its physiological changes. Nevertheless, the relationship between muscle fiber composition and intramuscular oxygenation is still unclear. In the present study we examined the differences in muscle fiber composition and their relationship to muscle oxygenation levels by comparing power lifters (Training group), who regularly engaged in muscular resistance training, with ordinary healthy people (Control group). The vastus lateralis muscle oxygenation level was measured via near infrared continuous wave spectroscopy, and muscle tissues were collected and used for histochemical analyses in order to calculate muscle fiber compositions. When the rate of decrease in muscle oxygenation after performing a single squat at 50 and 80% of the participants’ maximal lifting weight (MLW) was examined, the rate of decrease was significantly higher at 80% of MLW than at 50% of MLW in Training group, and was significantly higher in the Training group than in the Control group. Moreover, after performing multiple squat movements at 80% of MLW, compared with the Control group, the Training group showed a significant delay in the time it took for their muscle oxygenation level to recover to 50% of its original level at resting time (T1/2). A significant correlation between the occupancy of Type IIa fiber and T1/2, or between an average cross-sectional area of muscle fibers and T1/2 was noted. The present study demonstrated that in the Training group, in a cross-sectional area of Type IIa fiber that had increased, the decreased muscle oxygenation level due to performing squat exercises exhibited a delay in recovery at the resting time, suggesting that the amount of oxygen consumption was increased in the Training group because the crosssectional area of Type IIa fiber was larger, compared with the Control group.

A clinical statistical study was carried out on surgical correction of jaw deformation in patients who visited Osaka Dental University.1) The annual number of the new patients with jaw deformation slightly increased. A marked increase was observed after 1990.2) The male/female ratio in the new patients was about 1: 2.3) The mean age at the time of initial consultation was 20.4 years for males and 20.3 years for females.4) According to age groups, the number of patients aged 16-20 years accounted for 47.6% and that of patients aged 21-25 years for 30% of all patients.5) Concerning area distribution in the new patients, Osaka prefecture accounted for the highest percentage (71%), followed in order by Hyogo, Nara, and Oyoto prefectures.6) According to the type of jaw deformation, skeletal mandibular protrusion accounted for the highest percentage (84%).

Expression and implication of carbohydrate antigens in squamous cell carcinomas (SCCs) in oral cavity was examined. In the cell lines, type 2H and Lewis y antigens were markedly expressed. In the tissues from SCC patients and benign disorders, type 2H was highly expressed in hyperplasia (96.4 %), displasia (92.9 %) and SCC (100 %). Lewis y was, in turn, expressed mainly in cancer tissues (91.3 %), suggesting that Lewis y is a cancer-associated antigen. Normal oral mucosa showed no expression of these blood group antigens. Surprisingly, Lewis y antigen disappeared in the invasion sites where Ki-67 was definitely stained. Over-expression of Lewis y with manipulation of a fucosyltransferase cDNA resulted in suppression of cell growth and invasion, and knockdown of Lewis y also brought about increased cell growth and invasion. In either situations, no changes in the expression of sialyl-Lewis x could be found. Lowered tumor growth and invasion into surrounding tissues were also shown in Lewis y-positive SCC grafts in nu/nu mice. All these results together with alternative staining between Lewis y and Ki-67 in cancer tissues and FUT1 transfectants suggested that loss of Lewis y is a crucial event for the late stage of SCCs.

Ganglioside GD3 is specifically expressed in human melanomas, and plays a role in the enhancement of malignant phenotypes of melanoma cells. To analyze the mechanisms by which GD3 enhances malignant properties and signals in melanomas, it is essential to clarify how GD3 interacts with membrane molecules on the cell membrane. In this study, we performed proteomics analysis of glycolipid-enriched microdomains (GEM) with current sucrose density gradient ultracentrifugation of Triton X-100 extracts and MS. We also examined GD3-associated molecules using enzyme-mediated activation of radical sources (EMARS) reaction combined with MS. Comparison of molecules identified as residents in GEM/rafts and those detected by EMARS reaction using an anti-GD3 antibody revealed that a relatively low number of molecules is recruited around GD3, while a number of membrane and secreted molecules was defined in GEM/rafts. These results suggested that EMARS reaction is useful to identify actually interacting molecules with gangliosides such as GD3 on the cell membrane, and many other microdomains than GD3-associating rafts exist. Representative examples of GD3-associated molecules such as neogenin and MCAM were shown.

Aluminum nitride powder was synthesized from aluminum polynuclear complexes. Basic aluminum chloride (BAC) provided the aluminum polynuclear complexes. The effect of the milling treatment for the precursor made from basic aluminum chloride and glucose on the particle size was investigated. BAC and glucose were dissolved in water and mixed homogeneously. AIN powder was obtained by calcining under a nitrogen gas flow after drying, milling by vibration mill, and precalcining at 800°C. Then excess carbon was removed by firing in air. It was found that the milling treatment affected the particle size of AIN powder and nitridation mechanism. Without the milling treatment, AIN powder was synthesized directly from the γ-alumina of an intermediate product. In using a milled precursor, however, α-alumina was formed during the calcining, and the particle size of the AIN powder obtained was about five times larger than that of AIN powder synthesized from a raw precursor. The formation of α-alumina phase began at the rather low temperature of 800°C. These results suggest that the mechanochemical effects added by the milling promotes the formation of α-alumina during calcining, and the α-alumina phase formed accelerates the particle growth. AIN powders obtained were very uniform. The oxygen content and the surface area of AIN powders synthesized from the raw and milled precursors were 2.9 wt% and 17.5 m2/g, and 1.1 wt% and 3.6 m2/g, respectively.

AIN powder was synthesized from aluminum polynuclear complexes. Basic aluminum chloride and basic aluminum lactate were used as the aluminum polynuclear complexes. These starting materials and glucose were dissolved in water and mixed homogeneously. AIN powder was obtained by calcining after drying and precalcining at 800°C under nitrogen gas flow. Then excess carbon was removed by firing in air. Nitridation in the system was investigated and compared with that in the alumina–carbon black system. It was found that in our reaction system nitridation began and proceeded at lower calcination temperatures above 1200°C than in the alumina–carbon black system. Using aluminum polynuclear complexes, AIN was synthesized through the nitridation of γ-alumina and produced in a very fine and sharp particle size distribution.

The sintering behavior of fine AIN powder synthesized from an aluminum polynuclear complex was investigated. The focus of this work was to investigate the densification behavior of the AIN powder with different particle sizes (specific surface area: 3.2–22.8 m2/g). The AIN powder was synthesized from basic aluminum chloride and glucose mixed in a water solution. This powder was divided into two groups: one with 2 wt% Y2O3 added as the sintering aid and the other without such an additive. The AIN powder investigated possessed favorable densification potential. The density of the AIN powder with a surface area of 16.6 m2/g and without additives attained theoretical density at 1700°C. Adding Y2O3 further decreased the sintering temperature required for full densification to 1600°C. It is speculated that low-temperature sintering of our fine AIN powder with Y2O3 proceeds in two steps: in the initial stage, sintering proceeds predominantly through interdiffusion between yttrium aluminates formed on the AIN powder surface; in the second stage, the densification may occur by the interdiffusion between solid phases formed by a reaction between the yttrium aluminates and AIN. To investigate the effect of oxygen on sintering, the content of oxygen in AIN powder was varied while the particle size was kept constant. In this study, the difference in surface oxygen content scarcely affected the sintering behavior of fine AIN powder.

AIN powder was synthesized from aluminum polynuclear complexes. Basic aluminum chloride and basic aluminum lactate were used as the aluminum polynuclear complexes. These starting materials and glucose were dissolved in water and mixed homogeneously. AIN powder was obtained by calcining after drying and precalcining at 800°C under nitrogen gas flow. Then excess carbon was removed by firing in air. Nitridation in the system was investigated and compared with that in the alumina–carbon black system. It was found that in our reaction system nitridation began and proceeded at lower calcination temperatures above 1200°C than in the alumina–carbon black system. Using aluminum polynuclear complexes, AIN was synthesized through the nitridation of γ-alumina and produced in a very fine and sharp particle size distribution.

Effect of carbon on the synthesis of AlN powder from aluminum polynuclear complexes was investigated. Aqueous solutions of basic aluminum chloride and glucose were used as starting materials. C/Al molar ratio was varied to investigate the effect on the reactivity of nitridation and particle size of synthesized AlN powder. The structure of the solution was investigated by 27Al and 13C-NMR spectroscopy. It was found that reactivity of nitridation took a maximum value at 1.8-2.0 of C/Al molar ratio and thereafter decreased gradually. This indicates that excess carbon inhibits the transportation of reaction gases at a gap among the particles. In this reaction system, C/Al molar ratio for the maximum reactivity nitridation is much smaller than that in the alumina-carbon black mixture system. Furthermore, this value is closer to the theoretical value for a complete nitridation reaction. These results suggest that carbon in our system is distributed more homogeneously than that in the alumina-carbon black mixture system. The particle size of the synthesized AlN powder decreases with increasing C/Al molar ratio. Obviously, carbon has a strong effect on the particle size of γ-alumina, which is formed as an intermediate in the nitridation process, and prevents the grain growth of the synthesized AlN particles. It was found that aluminum polynuclear complex and glucose formed a precursor in the solution.

We have previously shown (J. Exp. Med. 1984. 160: 1170) that murine B cells activated with lipopolysaccharide (LPS) and rabbit anti-mouse Ig antibodies together (LPS-RaMIg blasts) express high-affinity interleukin 2 receptors (IL 2R) and respond to IL 2. This is not the case for B cells activated with either LPS alone (LPS blasts) or anti-Ig alone. In the present study IL 2R function and expression were further investigated by using this model. First, it was found that LPS-RaMIg blasts internalize IL 2 in a time- and temperature-dependent manner very similar to that occurring in CTLL (T lineage) cells. LPS blasts, however, did not internalize IL 2. LPS blasts were found to express 12 times less binding sites for anti-IL 2R monoclonal antibody (PC 61 monovalent Fab) as compared to LPS-RaMIg blasts and at least 30 times less IL 2 binding sites of high as well as of lower affinity. Second, with regard to the requirements for receptor expression, it was observed that either anti-Ig or phorbol diester (phorbol-12-myristate 13-acetate) can induce IL 2R and IL 2 responsiveness (proliferation assay) in LPS blasts but not in fresh B cells. Taken together these results provide further evidence for the similarity of IL 2R function in activated B and T cells, confirm that surface-Ig cross-linkage and phorbol-12-myristate 13-acetate have similar effects on B cells and suggest that LPS on the one hand, and phorbol-12-myristate 13-acetate or anti-Ig on the other provide complementary signals necessary for IL 2R expression.

In a recent report from our laboratory (J. Exp. Med. 1984. 160: 1070), it has been shown that B cells which have been activated with lipopolysaccharide (LPS) plus anti-Ig antibodies express interleukin 2 (IL2) receptors and proliferate in response to pure IL2. In the present study, we have investigated the role of IL2 during proliferative B cell responses supported by various T cell supernatants (SN) as well as by LPS itself. The following results were obtained: (a) B cells activated with LPS plus anti-Ig were found to proliferate in response to either recombinant IL2, T cell SN or fresh LPS; (b) a monoclonal anti-IL2 receptor antibody (PC61) could completely inhibit the effects of recombinant IL2 or various T cell SN (cloned T helper cell SN, EL4 SN, concanavalin A-spleen cell SN or mixed leukocyte culture SN), but did not interfere with the effect of LPS; (c) B cells activated with LPS or LPS plus anti-Ig were not found to secrete detectable amounts of IL2 by themselves. Thus, the data demonstrate that LPS-stimulated B cell proliferation requires neither B cell autocrine nor T cell-derived IL2. On the other hand, with regard to LPS plus anti-Ig-activated B cells, IL2 appeared to be the only growth-promoting activity that could be detected in the supernatants obtained from total normal spleen cell populations.

In this study we show that a rapid colorimetric determination of alkaline phosphatase (APase) activity can be used in conjunction with a multiwell scanning photometer to quantitatively measure lymphokine-dependent B cell proliferation. Despite the fact that only activated but not resting B cells exhibit APase activity, i.e., that APase expression in B cells is variable, the results obtained by monitoring the enzyme activity were very similar to those obtained by measuring thymidine incorporation in two murine B cell systems. With regard to a potential application to the screening of the effects of hybridoma supernatants on B cells, it is relevant that HAT medium has practically no influence on the colorimetric B cell proliferation assay. Another advantage of the method is that since T cells and macrophages lack APase activity, one can detect the proliferation of B cells even in the presence of a high number of such cells, for example, in studying a T cell-dependent response.

The antibody against putative TRF-acceptor site(s) on B cells was raised in (DBA/2Ha × BALB/c) (DC)F 1 male mice (TRF-low-responders, putatively TRF acceptor site(s)-negative) against activated B cells from BALB/c mice (TRF-high-responders). Neonatal injection of this antibody resulted in a selective suppression of development of B cells responsive to TRF, whereas that of B cells capable of interacting directly with helper T cells through antigen was not affected by such treatment. Likewise, treatment of DNP-primed splenic B cells with biotinated DCF 1 anti-BALB/c B-cell antibody followed by treatment with avidin-ricin A conjugates could eliminate antibody-reactive B cells and remaining B cells did not respond to TRF, whereas the activity of the B cells responsive to cognate interaction with helper T cells was intact. These results clearly indicate that B cell subpopulation responsive to TRF is distinct from the other B cell subpopulation cooperating directly with helper T cells via antigen.

A 71-year-old woman with symptomatic Rathke's cleft cyst is reported. She had visual disturbance, diabetes insipidus and hypopituitarism. Imaging studies with computed tomography and magnetic resonance image revealed the presence of suprasellar mass. After the transsphenoidal surgery and treatment with DDAVP, her symptoms showed an improvement.

By using purified TRF and BCGF from two T cell hybridomas, B151K12 and FS6-14.13, genetic control of responsiveness of B cells to those respective factors was analyzed. Results revealed that B cells from DBA/2Ha mice were low-responders to TRF but not to BCGF. In contrast, B cells from CBA/N mice were low-responders to BCGF. In both cases, low responsiveness was inherited in X-linked recessive manner.

It is widely accepted that B cells possess two distinct receptors. One is the surface immunoglobiilin that recognizes the antigenic determinant; the other transmits the signals for proliferation and differentiation of the B cells after receiving differentiation stimuli mediated by a soluble factor derived from helper T cells. Previously1–4, we demonstrated that the responsivity of B cells to T cell-replacing factor (TRF) was under X-chromosome control, as determined by genetic analysis of the TRF-nonres-ponsiveness of B cells from DBA/ 2Ha mice. An X-linked B-cell defect of DBA/2Ha mice to TRF seemed to be due to the absence of an acceptor site(s). We have immunized the defective (DBA/2Ha♀×BALB/c♂)F1 male mice with antigen-primed parental BALB/c splenic B cells. The resulting alloantiserum preferentially inhibited the B-cell response mediated in vitro by TRF added continuously to the culture. We report here that this antiserum, which presumably contains antibody against the putative TRF-acceptor site, augments primary IgM responses to simultaneously administered suboptimal doses of heterologous erythrocytes injected into TRF high-responder animals. These augmented IgM-PFC (plaque -forming cell) responses were also observed in other X-Iinked B-cell defective CBA/N mice, but not in TRF low-responder DB A/2Ha mice, suggesting that the putative TRF-acceptor site(s) functionally detectable by this antiserum is distinct from the Lyb 3 and Lyb 5 components.