Search

The regeneration of wounded stratified epithelium is accomplished via the migration of keratinocytes from the margins of the wound. However, the process of keratinocyte migration on the wound surface and the role of epithelial stem cells during re-epithelialization remain to be elucidated. Therefore, we administered BrdU to embryonic mice and generated epithelial defects on the buccal mucosa of these mice at two weeks after birth, using CO(2) laser irradiation, with which we removed the entire thickness of the epithelium. In the unwounded epithelium, cytokeratin 14, p63, and BrdU were localized within the basal layer of the epithelium, but the majority of cells within the regenerated epithelium were immunopositive for these proteins. PCNA-negative and BrdU-positive basal keratinocytes, which evidence a slow cell cycle, were localized solely within the basal layer of the unwound epithelium facing the tips of dermal papillae. After laser irradiation, these basal keratinocytes facing the tips of the papillae evidenced positive immunoreactivity for PCNA, in addition to BrdU. These results indicate that epithelial stem cells of oral mucosa may be localized in the basal layer of the epithelium facing the tips of dermal papillae, and may migrate laterally with other basal keratinocytes in response to external stimuli.

We investigated the expression and localization of connexins (CX) 26 and 43 in the rat gingival epithelium. RT-PCR analysis revealed CX26 gene expression in both the upper and lower layers of the gingival epithelium and in the total epithelial layer, whereas CX43 gene expression was limited to the lower layer and the total epithelial layer. Immunoreactivity for CX43 was observed in the membranes of adjacent cells from the basal layer to the middle of the prickle cell layer, while immunoreactivity for CX26 was observed in the granular cell layer and lower part of the squamous cell layer. Merged images revealed the co-localization of CX26 and CX43 in the middle of the prickle cell layer. By immuno-electron microscopy, gap junctions appeared curved, hemi-circular, or annular within the cytoplasm, and gold particles indicating the presence of CX43 were localized at the outer edges of these cytoplasmic formations. These results suggest that CX43 is associated with the regulation of cell proliferation and that increased CX26 expression is associated with differentiation of keratinocytes. Thus, degradation of CX43 is considered to play an essential role in differentiation of the rat gingival epithelium.

Many morphological and developmental studies have demonstrated the characteristics of tight junctions (TJs) between odontoblasts. However, detailed localization of TJ-associated proteins in odontoblasts and their functions has not yet been clarified. To elucidate the relationship between the establishment of TJ structures and the differentiation of odontoblasts during early dentinogenesis, we studied the expression and localization of constituent proteins of TJs (claudin-1, occludin, ZO-1 and ZO-2) between odontoblasts in rat lower incisors using Western blotting, immunofluorescence and immunoelectron microscopy. When the expression of claudin-1 increases at the distal portion of mature odontoblasts, the TJs form complex networks of strands, and odontoblasts differentiated by developing distal membrane domains and by secreting specific molecules for mineralization. We conclude that the TJs of odontoblasts may play an important role in the differentiation of odontoblasts in rat lower incisors during early dentinogenesis.

The submandibular gland (SMG) is a tissue that can be regenerated in a tissue injury model and that has adult stem cells capable of self-renewal and differentiation into functional cells. We have analyzed the localization of label-retaining cells (LRCs), which are putative progenitor cells, by using the BrdU-labeling method. 5-Bromo-2'-deoxyuridine (BrdU) injection followed by a long chasing period permitted the identification of LRCs based on the slow-cycling characteristic. In order to confirm the accurate localization of LRCs, BrdU and SMG-specific markers, including aquaporin5, cytokeratin, and smooth muscle actin, were examined by double-immunofluorescence staining. We found that LRCs were distributed in the acinus, duct, myoepithelium, and connective tissue. Moreover, ABCG2 (a known stem cell marker) was used for the characterization of LRCs and the localization of cells as putative stem/progenitor cells. ABCG2-expressing cells were distributed in various regions of the SMG but did not co-localize with LRCs. We suggest that putative progenitor cells exist in various regions of the SMG and have diverse capacities to differentiate into specific cells.

We herein report on a Japanese boy with infantile neuronal ceroid lipofuscinosis (INCL). He was born of incest to a girl and her maternal uncle. His development was normal at 12 months, and began to display regression at 14 months. He lost his social smile and tracking eye movement at 16 months, and could not stand and developed severe hypotonic tetraplegia at 19 months. Myoclonic movement was observed in his trunk, eye and extremities. His height, body weight and head circumstance had been normal. Both MRI and CT scans of his head showed severe cerebral, cerebellar and brainstem atrophy. The electroretinogram showed a decrease in amplitude. Enzyme studies revealed a deficiency of palmitoyl-protein thioesterase activity in his lymphocytes at 0.98 nmol/h/mg protein (control: 90.99+/-34.23). This is the first case of INCL in Japan diagnosed by enzyme activity deficiency.

Aquaporins (AQPs) are integral membrane proteins that function as channels for the transfer of water and small solutes across membranes. In mammalian cells, 13 isoforms (AQP0-12) have been identified, and these exhibit unique patterns of expression in various cell types and tissues. Among these isoforms, AQP6 is considered to function not as water channel, but as an anion channel. We investigated the presence and localization of AQP6 in rat parotid acinar cells. AQP6 mRNA was detected in these cells by using reverse transcription/polymerase chain reaction, and Western blotting analysis identified a protein band that reacted with an anti-AQP6 antibody in the membrane fraction and secretory granule membrane. In order to localize AQP6, we used the anti-AQP6 antibody for histological immunodetection. Under confocal microscopy, we observed positive immunoreactions near the tight junctions of parotid acinar cells. Immunolabeling of ultrathin cryosections detected AQP6 near tight junctions and around secretory granule membranes. Immunoelectron microscopy confirmed the presence of AQP6 in the membranes of isolated secretory granules. These results suggest that AQP6 participates in water and anion transport in plasma membranes near tight junctions and secretory granule membranes in rat parotid acinar cells.

Aim: To investigate the responses of cultured rat pulp cells to heat stress., Methodology: Pulp cells were obtained from rat incisors and cultured at 37 degrees C. The cells were cultured at 42 degrees C for 30 min and then cultured at 37 degrees C again. Morphology, alkaline phosphatase (ALP) activity and expression of heat shock protein 25 (HSP25) were investigated at 0, 1, 3, 5, 7, 10 and 14 days following stimulation. As a control, the cells were maintained at 37 degrees C., Results: Although there were few cells of apoptosis immediately after heat stress, there were mitotic cells from day 1 after heat stress. ALP activity in the heat stress group significantly increased at days 7 and 14 compared with the control group (about 1.7-fold, P < 0.01, Friedman test). HSP25 expression increased in both groups, with HSP25 in the heat stress group being expressed earlier than in the control group, and nuclear localization of HSP25 was observed at days 0 and 1 in heat-stressed cells., Conclusion: These results suggest that heat stress not only induces HSP25 but also enhances ALP activity in pulp cells.

We herein report a 16-year-old boy who presented with fever and generalized convulsion. His symptoms progressively worsened, and electroencephalography (EEG) showed status with bi-frontal focus. One month after the initial presentation, examination of the cerebrospinal fluid showed pleocytosis, an increase in the IgG index and a positive oligoclonal IgG band. Brain MRI suggested slightly high signal intensity in the cerebral gray matter on FLAIR images. We considered that the patient had autoimmune meningoencephalitis, and therefore treated him with methyl-predonisolone pulse therapy, which substantially improved both his clinical condition and EEG findings. Before the therapy, the IgM and IgG auto-antibodies against glutamate receptor (GluR) epsilon 2 and delta 2 were positive, however they became negative after the therapy. In conclusion, some cases of meningoencephalitis with auto-antibodies against GluR may show an electrical status, mild brain MRI findings, and a good response to methylprednisolone pulse therapy. These clinical features were quite different from those observed for cases of Rasmussen's encephalitis.

The effect of insulin-dependent diabetes mellitus (IDDM) on bone metabolism was evaluated using the streptozotocin (STZ)-induced diabetic rat 1 week after the induction of diabetes. The urinary excretion of cross-linked N-telopeptides of type I collagen (NTx) and deoxypyridinoline (Dpd) in diabetic rats increased to 3.6-fold and 1.2-fold the control level, respectively. The amount of hydroxyproline and calcium in the distal femur of diabetic rats significantly decreased to 76% and 90% of the control, respectively. The levels of serum osteocalcin and alkaline phosphatase (ALP) activity in the distal femur of the diabetic rats were significantly reduced to about 40% and 70% of the control levels, respectively. The decrease in the expression osteocalcin was observed in distal femur of the diabetic rats, although the level of ALP mRNA was unchanged. The activity and the mRNA level of tartrate-resistant acid phosphatase (TRAP) increased to 1.5- and 2.3-fold the control level, respectively, in distal femur of the diabetic rats. The activity, protein, and mRNA levels of cathepsin K of diabetic rats also elevated to about 2-, 2.3-, and 2-fold the control levels, respectively. These results suggest that IDDM contributes to bone loss through changes in gene expression of TRAP and cathepsin K in osteoclasts as well as osteocalcin in osteoblasts resulting in increased bone resorptive activity and decreased bone formation.

Pleomorphic adenomas (PAs) of salivary glands are characterized by the mixed appearance of epithelial and mesenchymal-like components such as myxoid and chondroid tissues. Although various studies have examined PAs, thus far it is not clear how PAs make these multiple components. Thus, clarification of the histodifferentiation of this unique salivary gland tumor using not only tissues in vivo but also PA cells cultured in vitro is necessary. However, no in vitro model of PA has been reported, because normal and benign tumor cells tend to grow slowly and senesce quickly in culture. Therefore, we immortalized cells using transfection of the hTERT gene without otherwise altering the nature of those cells. The immortalized PA cells expressed mRNA of the pleomorphic adenoma gene 1 and showed epithelial and neoplastic myoepithelial characteristics by immunohistochemical immunofluorescence analyses and ultrastructural study. Our findings suggest that these cells will be a useful model to study the cellular differentiation of PA.

Connexins are gap-junction proteins forming hexameric structures in the plasma membranes of adjacent cells, thereby creating intercellular channels. Connexin 43 (CX43) is expressed in pulp tissue. However, its function in dental pulp tissue has yet to be fully investigated. We have employed antisense oligonucleotides (AS) against rat CX43 to study the role of CX43 in dental pulp cells. Cultured dental pulp cells were treated with AS or sense (S) oligonucleotides. The number of cells in the AS-treated groups was approximately 1.3-fold that in the S-treated controls. Growth rates were significantly different between the AS- and S-treated groups at 48 h (P < 0.01). An alkaline phosphatase assay revealed that AS-treated pulp cells dramatically decreased at 48 h after AS incorporation, whereas S-treated pulp cells showed no marked changes. Western blot analysis revealed that heat-shock protein 25 was highly expressed in S-treated cells but was only weakly expressed in AS-treated cells at 48 h. Furthermore, AS-treated cells highly expressed CX45, whereas S-treated cells exhibited high expression of CX32. These results suggest that CX43 is involved in cell growth, mineralization, and differentiation to odontoblasts in rat pulp cells, and that CX43 plays the opposite role to that of CX45.

We herein report the findings of a 2-year-6-month-old boy, who had been experiencing monocular pendular nystagmus, strabismus, and episodic eye deviation nystagmus, intractable dystonia and apneic attack which all began when he was 2 days of age. He underwent a complete blood count test, blood chemistry test, analysis of amino acids in the blood and urine, analysis of pyruvate/lactate in blood and cerebrospinal fluid, head computed tomography and magnetic resonance imaging and no abnormal results were identified. His attacks were resistant to multiple antiepileptic and dopaminergic drugs. He showed transient left and/or right hemiplegia after nystagmus, dystonia and/or apneic attacks at 8-months of age with retardation in intelligence. We diagnosed him to have alternating hemiplegia of childhood (AHC). We were unsure how to deal with his attacks after he was discharged from the hospital, however, resuscitation with the ambu bag by his mother at home and the intravenous infusion of diazepam or thiamylal at the hospital together was proven to be an effective method for treating his severe apneic attacks. The effect of diazepam and amantadine on these attacks was transient, however, the administration of flunarizine with amantadine resulted in an improvement in his attacks. We therefore consider the administration of flunarizine to be essential for the effective treatment of AHC in this case.

Benzothiadiazole (BTH) is a so-called plant activator and protects plants from diseases by activating the salicylic acid (SA) signaling pathway. By microarray screening, we identified BTH- and SA-inducible WRKY transcription factor (TF) genes that were upregulated within 3 h after BTH treatment. Overexpression of one of them, WRKY45, in rice (Oryza sativa) markedly enhanced resistance to rice blast fungus. RNA interference-mediated knockdown of WRKY45 compromised BTH-inducible resistance to blast disease, indicating that it is essential for BTH-induced defense responses. In a transient expression system, WRKY45 activated reporter gene transcription through W-boxes. Epistasis analysis suggested that WRKY45 acts in the SA signaling pathway independently of NH1, a rice ortholog of Arabidopsis thaliana NPR1, which distinguishes WRKY45 from known Arabidopsis WRKY TFs. Two defense-related genes, encoding a glutathione S-transferase and a cytochrome P450, were found to be regulated downstream of WRKY45 but were not regulated by NH1, consistent with the apparent independence of the WRKY45- and NH1-dependent pathways. Defense gene expression in WRKY45-overexpressed rice plants varied with growth conditions, suggesting that some environmental factor(s) acts downstream of WRKY45 transcription. We propose a role for WRKY45 in BTH-induced and SA-mediated defense signaling in rice and its potential utility in improving disease resistance of rice, an importance food resource worldwide.

Neuronal activities have recently been reported to exhibit power-law scaling behavior. However, it has not been demonstrated that the power-law component can play an important role in human perceptual functions. Here, we demonstrate that the power spectrum of magnetoencephalograph recordings of brain activity varies in coordination with perception of subthreshold visual stimuli. We observed that perceptual performance could be better explained by modulation of the power-law component than by modulation of the peak power in particular narrow frequency ranges. The results suggest that the brain operates in a state of self-organized criticality, modulating the power spectral exponent of its activity to optimize its internal state for response to external stimuli.

We previously demonstrated a dominant IgG response against XAGE-1b antigen in a lung cancer patient by serological analysis of antigens by recombinant expression cloning (SEREX) analysis using a cDNA library from the autologous OU-LU-6 tumor cell line. In this study, we investigated recognition of XAGE-1b on OU-LU-6 tumor by the patient CD4-expressing tumor-infiltrating lymphocytes (CD4 TIL). The response of CD4 TIL obtained from malignant pleural effusion was determined against autologous OU-LU-6 tumor cells and XAGE-1b mRNA-transfected PHA-stimulated CD4 T-cells (T-APC) from healthy individuals sharing HLA-DR with the patient by performing IFNgamma secretion and ELISPOT assays. The patient CD4 TIL recognized OU-LU-6 in an HLA-DR-restricted manner and XAGE-1b mRNA-transfected T-APC derived from DRB1 *0901-sharing healthy donor (HD)1 and HD2, but not DRB1 *1101-sharing HD3 or HD4. Epitope analysis using 17 18-mer peptides with 12 overlapping amino acids revealed that the CD4 TIL recognized XAGE-1b 33-49. The findings suggest that the patient CD4 T-cells recognized the XAGE-1b 33-49-related epitope on autologous OU-LU-6 tumor cells in a manner restricted by DR *0901.

XAGE-1 is a cancer/testis (CT) antigen and has been shown to be immunogenic in lung cancer patients. Among 4 alternative splicing variants, XAGE-1a, b, c and d, XAGE-1b mRNA was dominantly expressed in cancer. In this study, we generated a XAGE-1b mAb, USO9-13. The B cell epitope recognized by the USO9-13 mAb was in the C-terminal region of the XAGE-1b protein and is also recognized by sera from patients with lung adenocarcinoma. Using USO9-13 and an anti-Flag mAb, we examined the translation products of the 4 transcripts. The XAGE-1a and b transcripts translated to the XAGE-1b protein. The XAGE-1c transcript possibly translated to 9- and 17-aa polypeptides. The XAGE-1d transcript translated to a protein consisting of 69 amino acids. Immunofluorescence analysis using USO9-13 mAb showed that the XAGE-1b protein is located in the nuclei of cells. Immunohistochemically, nuclear staining was heterogeneously observed in 25/47 lung adenocarcinomas, 1/12 hepatocellular carcinomas and 1/11 gastric cancers, but not in adjacent normal tissues. These findings suggested that XAGE-1b is a promising target molecule for a cancer vaccine against lung cancer.

Temporomandibular disorders (TMD) represent a group of chronic painful conditions in the masticatory musculature and temporomandibular joint. To examine possible changes in cortical machinery in TMD patients, we compared neuromagnetic signals evoked by cortical neurons between healthy subjects and TMD patients while they were carefully observing the video frames of jaw-opening movements performed by another person. During the movement observation task in the healthy subjects, we found cortical activation in the following sequence with left hemisphere dominance: (1) the occipitotemporal region near the inferior temporal sulcus (human homologue of MT/V5 in monkeys), (2) the inferior parietal cortex (IPC), and (3) the anterior part of the inferior-lateral precentral gyrus (PrCG). In the TMD patients, however, we found deficit or marked attenuation of the neuromagnetic responses in the PrCG and IPC, while the activity of the MT/V5 showed no differences from that in the healthy subjects. In addition, we could not find any differences in cortical magnetic responses between healthy subjects and TMD patients when they were observing palm-opening movements, indicating that cortical dysfunction associated with jaw-movement observation is specific phenomena in the patients of TMD. Thus the present study provides new neuropathological evidence that TMD patients exhibit dysfunction of recognition mechanisms in cerebral cortex during motor observation, and suggests that disturbance of cortical functions regulating visuomotor integration would play a crucial role in development as well as aggravation of TMD.

Background: Osteopontin (OPN) expression in squamous cell carcinoma (SCC) of the tongue has not been clearly elucidated., Methods: We selected 46 cases of tongue SCC and investigated the expression of OPN by immunohistochemical staining. The immunopositive reaction and score for each case were semiquantitatively evaluated., Results: Scores were significantly higher in carcinoma nests than in neighboring normal epithelium or epithelial dysplasia. The OPN was expressed clearly in the cytoplasm of carcinoma cells. In cases of early invasive carcinoma, in particular, expression of OPN showed a remarkable increase at the invasion front compared with the non-invaded regions. However, there was no significant correlation between expression of OPN in the primary tumor nest and lymphatic metastasis, recurrence, or survival rate., Conclusion: This suggests that OPN is a useful biomarker of early invasion by SCC in tongue.

The inhibitory effects of tyrosine kinase inhibitors including tyrphostins 25, 47 and 51 on Shiga toxin 1-induced cell death and p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation were examined in Vero cells. Tyrphostin 47 significantly inhibited Shiga toxin 1-induced cell death and p38 MAPK phosphorylation. In contrast, tyrphostins 25 and 51 had no significant effect on the Shiga toxin 1-induced responses. These data indicate that Shiga toxin 1-induced cell injury occurs through a pathway sensitive to tyrphostin 47, and the target molecule for tyrphostin 47 opens up new opportunities for pharmacological intervention against Shiga toxin-producing Escherichia coli infectious diseases.

The present study investigated the expression and distribution of osteopontin in the mouse major salivary glands. The level of osteopontin expression in the mouse submandibular gland was higher (12.7-fold) than that in parotid and sublingual glands at the mRNA level. By Western blot analysis, intense positive bands were seen at the predicted molecular mass (about 55 kDa) in all the major salivary glands, while an approximately 30 kDa band of osteopontin was detected only in the submandibular gland. Indirect immunofluorescent and immuno-electron microscopy analyses demonstrated the localization of osteopontin in the luminal (apical) membranes of acinar cells in all the salivary glands. Osteopontin was also localized at the lumen of acini in the submandibular gland. These results suggest that the expression of osteopontin in the submandibular gland is different from that in the parotid and sublingual glands and that osteopontin may be degraded in the mouse submandibular gland.

Konjak mosaic virus (KoMV) belongs to the genus Potyvirus, family Potyviridae. The complete nucleotide sequence of KoMV F isolate (KoMV F) was determined. The genome is 9,544 nucleotides long excluding the 3' terminal poly A tail and encodes a typical potyviral 350-kDa polyprotein of 3,087 amino acids. Phylogenetic analysis using known potyvirus polyproteins shows that KoMV constitutes a branch with yam mosaic virus, close to another branch including Japanese yam mosaic virus, turnip mosaic virus, scallion mosaic virus and lettuce mosaic virus. The 3' terminal 1,842 nucleotides of a different isolate of KoMV, K-2, was also determined, covering the C-terminal 292 amino acids of the nuclear inclusion protein b (NIb), coat protein (CP), and the 3' untranslated region. The amino acid sequences of the KoMV F CP and the nucleotide sequences of the KoMV F 3' untranslated region showed 92.5 and 90.5% identity to the corresponding genes of K-2, 88.7-96.8 and 92.7-94.4% to those of Zantedeschia mosaic virus (ZaMV) isolates, 87.5-89.7% and 85.5-90.3% to those of Japanese hornwort mosaic virus (JHMV) isolates. These results showed that KoMV is a distinct potyvirus and that KoMV, ZaMV, and JHMV are members of the same potyvirus species. Considering that KoMV was the first of these to be described, ZaMV and JHMV may be considered isolates of KoMV.

Background and Objective: It is known that epithelial islands are embedded in the cementum during tooth root formation, but details of this process remain unknown. The purpose of this study was to investigate the dynamic characteristics of Malassez's epithelial rest cells in the cementum during tooth root formation in pigs in vivo., Material and Methods: The first molars of 6-mo-old pigs were used in this study. Specimens were decalcified before being embedded in paraffin. Paraffin sections were investigated using TdT-mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemical, and ultrastructural techniques., Results: Malassez's epithelial rest cells were located close to the root surface at the apical one-third of the periodontal ligament, and epithelial clusters surrounded by distinct lamina cementia were sometimes observed in the cementum. TUNEL-positive cells were detected only in the cementum. Malassez's epithelial rest cells in the periodontal ligament were completely surrounded by basement membranes, but epithelial clusters in the cementum were only intermittently surrounded by such membranes. Cytokeratin-positive cells in the superstratum of the cementum were directly connected by cementocytes and by desmosome-like structures. However, organelles were scarce in the cytokeratin-positive cells in the substratum of the cementum, and the matrix of the cementum was deposited in the cells., Conclusion: These results suggest that the majority of the fragmented Hertwig's root sheath remains in the periodontal ligament and that some cells, which are connected to cementoblasts, are embedded in the cementum and progress to apoptosis.

To investigate the relationship between the expression of connexin in basaloid squamous cell carcinomas (BSCC) and their rapid proliferation and invasive potential, we examined the effect of overexpression of connexin 43 (Cx43) in a BSCC-derived cell line (BSC-OF). BSC-OF was transfected with Cx43 to obtain 15 clones with a stable expression of Cx43. In these cells, although Cx43 was distributed throughout the cytoplasm, it did not form connexon plaque. In almost all of the clones, cell proliferation was clearly suppressed. Furthermore, we investigated cell migration and invasion in three clones that showed a remarkable down-regulation in cell growth, and found that Cx43 transfection showed no significant effect on either. These results suggest that Cx43 plays a role as a tumor suppressor in the cytoplasm of Cx43-transfected BSC-OF cells. However, no definite correlation was found between Cx43 and cell migration and invasion.

Adsorption of antimicrobial peptide histatin 5 on a poly(methyl methacrylate) denture base may serve to prevent biofilm formation, leading to a reduction of denture-induced stomatitis. This study focused on adsorption behavior of histatin 5 onto PMMA surfaces modified using a cold plasma technique and the effectiveness of histatin 5 adsorption for reducing Candida albicans biofilm formation by the quartz crystal microbalance-dissipation (QCM-D) technique. PMMA spin-coated specimens were treated with oxygen (O(2)) plasma using a plasma surface modification apparatus. The amount of histatin 5 adsorbed onto the PMMA treated with O(2) plasma is more than six times greater than that adsorbed onto untreated PMMA. The degree of histatin 5 adsorption had a negative correlation with the contact angle, whereas that of zeta-potential showed no significant correlation. XPS analysis revealed that the introduction of the carboxyl and O(2) functional groups were observable on the O(2) plasma-treated PMMA. Increased surface hydrophilicity and the formation of the carboxyl could be responsible for histatin 5 adsorption on plasma-treated PMMA. There is no significant difference between histatin-adsorbed PMMA and control PMMA for C. albicans initially attached. On the contrary, the amount of C. albicans colonization on histatin-adsorbed PMMA was significantly less than the control.

Purpose: NY-ESO-1 belongs to a class of cancer/testis antigens and has been shown to be immunogenic in cancer patients. We synthesized a complex of cholesterol-bearing hydrophobized pullulan and NY-ESO-1 protein (CHP/ESO) and investigated the in vitro stimulation of CD8 and CD4 T cells from peripheral blood mononuclear cells in healthy donors with autologous CHP/ESO-loaded dendritic cells as antigen-presenting cells., Experimental Design: In vitro stimulation of CD8 or CD4 T cells was determined by IFNgamma ELISPOT assays against autologous EBV-B cells infected with vaccinia/NY-ESO-1 recombinant virus or wild-type vaccinia virus as targets and by ELISA measuring secreted IFNgamma., Results: NY-ESO-1-specific CD8 and CD4 T cells were induced. In a donor expressing HLA-A2, CD8 T cells stimulated with CHP/ESO-loaded dendritic cells recognized naturally processed NY-ESO-1(157-165), an HLA-A2-binding CD8 T cell epitope. NY-ESO-1 CD4 T cells were Th1-type. We identified a new HLA-DR15-binding CD4 T cell epitope, NY-ESO-1(37-50)., Conclusions: These findings indicate that CHP/ESO is a promising polyvalent cancer vaccine targeting NY-ESO-1.

We examined expression of syndecan-1 in squamous cell carcinoma (SCC) of tongue using immunohistochemistry. Forty-three cases of SCC arising in lateral border of tongue were investigated. From the immunohistochemical staining pattern, the cases were divided into two groups based on expression of syndecan-1 at the supra-peripheral cells of the tumor nest: Group A, completely or mainly positive; Group B, sporadically positive or negative. Most poorly differentiated SCC cases were classified into Group B (81.8%). The number of Group B cases in T1-2 was different from that in T3-4. The number of cases where syndecan-1 expression was reduced was much greater in T3-4, and represented the majority of Group B (86.7%). More than 80% of Grade 4D cases were in Group B (83.3%) based on the Yamamoto-Kohama criteria. These results indicate that reduction of syndecan-1 correlates to histological grade, tumor size and mode of invasion in tongue SCC.

Simvastatin acid (SVA) has been reported to stimulate bone formation with increased expression of BMP-2. Therefore, immobilization of SVA onto dental implants is expected to promote osteogenesis at the bone tissue/implant interface. The aim of this study was to evaluate the immobilization behavior of SVA onto titanium (Ti), O(2)-plasma treated titanium (Ti + O(2)), thin-film coatings of hexamethyldisiloxane (HMDSO), and O(2)-plasma treated HMDSO (HMDSO + O(2)) by using the quartz crystal microbalance-dissipation (QCM-D) technique. HMDSO surfaces were activated by the introduction of an OH group and/or O(2)-functional groups by O(2)-plasma treatment. In contrast, titanium surfaces showed no appreciable compositional changes by O(2)-plasma treatment. The QCM-D technique enabled evaluation even at the adsorption behavior of a substance with a low molecular weight such as simvastatin. The largest amount of SVA was adsorbed on O(2)-plasma treated HMDSO surfaces compared to untreated titanium, HMDSO-coated titanium, and O(2)-plasma treated titanium. These findings suggested that the adsorption of SVA was enhanced on more hydrophilic surfaces concomitant with the presence of an OH group and/or O(2)-functional group resulting from the O(2)-plasma treatment, and that an organic film of HMDSO followed by O(2)-plasma treatment is a promising method for the adsorption of SVA in dental implant systems.

Objectives: The aim of this study was to investigate the differential immunolocalization of laminin gamma(2) and integrin beta(4) in primary cultures of the rat gingival epithelium., Methods: The gingival epithelium was obtained from Sprague-Dawley rats and was cultured in serum-free keratinocyte growth medium (DK-SFM). Western blotting analysis, immunofluorescence, confocal laser scanning microscopy (CLSM), and immuno-gold labeling for laminin gamma(2) and integrin beta(4) were employed. CLSM images for laminin and integrin were analyzed in horizontal (x-y axis) and in vertical (x-z axis) sections., Results: Both laminin gamma(2) and integrin beta(4) were detected by Western blot analysis in the gingival epithelium. Immunolocalization of laminin gamma(2) was distinct in the cytoplasm to form one or two irregular rings in gingival epithelial cells. By contrast, integrin beta(4) was localized diffusely in the cytoplasm. F-actin (indicating actin filaments) was clearly discernible at the periphery of the cytoplasm to form a cellular fringe. In x-z axis images obtained by CLSM, laminin gamma(2) was recognized as large foci in the most inner portion just above the basal plasma membrane. Integrin beta(4) existed in the area where F-actin was labeled surrounding the membrane. Immuno-electron microscopy showed that 10nm colloidal gold particles indicating laminin gamma(2) were mainly localized at the extracellular portion and in the peripheral cytoplasm, whereas integrin beta(4) was distributed in the cytoplasm close to the basal plasma membrane but not in extracellular regions., Conclusions: In primary cultures of the rat gingival epithelium, both laminin gamma(2) and integrin beta(4) may be produced by the epithelium, and irregular rings of laminin gamma(2) are formed in areas where gingival cells adhere to the extracellular matrix.

Previous reports have shown the expression of several mechanosensitive ionic channels on the plasma membrane in odontoblasts, which are the cells responsible for dentin formation. The membrane characteristics of odontoblasts imply that they could play critical roles in the mechano-transduction of fluid displacement within dentinal tubules into the electrical cell signals, to carry dentin sensation to the central nervous system. However, the direct ionic mechanism underlying such a dentin nociceptive function remains unclear. In the present study, we investigated the expression of the transient receptor potential vanilloid subfamily member 1 (TRPV1) channel--which essentially contributes to the detection of pain sensation--in rat odontoblasts by immunohistochemical and nystatin perforated patch-clamp techniques. Immunohistochemical observation showed the localization of TRPV1-immunoreactions on the distal regions of odontoblast membranes. In the patch-clamp experiments, we observed capsaicin-induced inward currents that were inhibited by capsazepine, a TRPV1 channel antagonist. Our results indicate a significant expression of TRPV1 channels in odontoblasts, suggesting that odontoblasts may directly respond to noxious stimuli such as a thermal-heat stimulus, and point to the necessity for a reconsideration of the cellular mechanisms of dentin sensation based on the transmembrane ionic signals in odontoblasts.

Aim: To investigate the expression of osteocalcin mRNA in young and in aged human dental pulp tissue to determine the characteristics of osteocalcin expression., Methodology: Human dental pulp tissues of the third molars were obtained from healthy young (17-23 years) and aged (>50 years) subjects, and total RNA was extracted. Osteocalcin mRNA expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between the value in young and aged pulp was statistically analysed., Results: Osteocalcin mRNA was detected in all samples of human dental pulp tissue homogenates by RT-PCR analysis. Osteocalcin mRNA was expressed in young adult dental pulp but was decreased in aged human dental pulp. QRT-PCR analysis also showed a reduced expression of osteocalcin mRNA in aged human pulp. Expression of osteocalcin in young human pulp was significantly higher (about sixfold) than in aged pulp (P<0.01, Mann-Whitney U-test)., Conclusion: Reduction of osteocalcin expression may be associated with the loss of viability in human dental pulp tissue, and may be a characteristic of aged human dental pulps.

Laminin-5 (Ln-5) is an important molecule associated with epithelial cell adhesion and migration. In the gingiva around the tooth, Ln-5 localizes within basement membranes between the junctional epithelium (JE) and the tooth or connective tissue. Recently, we reported that in the oral mucosa around a dental implant, Ln-5 is expressed within the basement membranes at the implant-peri-implant epithelium (PIE) interface, and at the PIE-connective tissue interface. However, the ultrastructural localization of Ln-5 within or along the PIE has not yet been reported. Therefore, peri-implant oral mucosa was treated with anti-Ln-5 (gamma2 chain) antibody and examined using immuno-electron microscopy. Ln-5 was localized in the cells of the innermost-third layer and basal layer of the PIE. A 100-nm-wide Ln-5-positive internal basal lamina (basement membrane) and hemidesmosomes as adhesion structures were formed at the apical portion of the implant-PIE interface. However, at the upper-middle portion of the interface, these adhesion structures were not observed. Furthermore, at the PIE-connective tissue interface, the Ln-5-positive external basal lamina (basement membrane) and hemidesmosomes were partially deficient. Judging from these findings, we concluded that Ln-5 contributes to the attachment of the PIE to the titanium surface, and that PIE attached to titanium at the apical portion of the dental implant-PIE interface.

Aquaporin-11 (AQP11) has been identified with unusual pore-forming NPA (asparagine-proline-alanine) boxes, but its function is unknown. We investigated its potential contribution to the kidney. Immunohistochemistry revealed that AQP11 was localized intracellularly in the proximal tubule. When AQP11 was transfected in CHO-K1 cells, it was localized in intracellular organelles. AQP11-null mice were generated; these mice exhibited vacuolization and cyst formation of the proximal tubule. AQP11-null mice were born normally but died before weaning due to advanced renal failure with polycystic kidneys, in which cysts occupied the whole cortex. Remarkably, cyst epithelia contained vacuoles. These vacuoles were present in the proximal tubules of newborn mice. In 3-week-old mice, these tubules contained multiple cysts. Primary cultured cells of the proximal tubule revealed an endosomal acidification defect in AQP11-null mice. These data demonstrate that AQP11 is essential for the proximal tubular function. AQP11-null mice are a novel model for polycystic kidney diseases and will provide a new mechanism for cystogenesis.

Objective: It is still an open question whether cells directly attached to the tooth (DAT) cells are migratory or non-migratory cells. The purpose of this study was to examine cytoskeletal and surface structures of DAT cells that might be involved in migration., Methods: We investigated the distribution of stress fibers composed of actin filaments in DAT cells using phallacidin fluorescent dye methods in a confocal laser scanning microscope. To observe the three-dimensional structure of the DAT cell surface, the osmium maceration scanning electron microscope (SEM) method, which removes various soluble materials between DAT cells and the enamel, was employed., Results: Stress fibers were found in the most apically located DAT cells, and were arranged in parallel to the presumable cervical-line, whereas some of the fibers ran parallel to the tooth axis in the more coronally located DAT cells. The parallel arrangement to the tooth axis of the fibers may be involved with migration for turnover, and the parallel accumulation to the presumable cervical-line may be concerned with the cervical contraction of DAT cells. Osmium maceration SEM images at high magnification revealed the existence of microvilli-like structures on the enamel surfaces (facing to the tooth surface) of DAT cells after removal of the soluble matrices. The thicknesses of the microvilli-like structures on the enamel surfaces and cell processes of intercellular bridges were significantly different., Conclusion: DAT cells possess stress fibers arranged in parallel to the tooth axis and to the presumable cervical-line in the cytoplasm, and microvilli-like structures on their enamel surfaces. These results suggest that these structures contribute to DAT cell migration.

Purpose: XAGE-1 was originally identified by the search for PAGE/GAGE-related genes using expressed sequence tag database and was shown to exhibit characteristics of cancer/testis-like antigens. Four transcript variants XAGE-1a, XAGE-1b, XAGE-1c, and XAGE-1d have been identified thus far. We recently identified XAGE-1b as a dominant antigen recognized by sera from lung adenocarcinoma patients. We here investigated the mRNA expression of four XAGE-1 variants and XAGE-1 protein expression in non-small cell lung cancer (NSCLC). Humoral immune response to XAGE-1b was also evaluated in patients., Experimental Design: Forty-nine NSCLC specimens were analyzed for the expression of four XAGE-1 transcript variants by conventional 30-cycle and real-time reverse transcription-PCR and XAGE-1 protein expression by immunohistochemistry. Sera from 74 patients were analyzed for XAGE-1b antibody production by ELISA and Western blot., Results: XAGE-1b and XAGE-1d mRNA were detected in 15 and 6 of 49 lung cancer specimens, respectively. No XAGE-1a or XAGE-1c mRNA expression was observed. XAGE-1b mRNA expression was observed in 14 of 31 (45%) adenocarcinoma and 1 of 18 (6%) lung cancer with other histologic types. Immunohistochemical analysis using a XAGE-1 monoclonal antibody showed that 14 of 15 XAGE-1b mRNA-positive and 3 of 34 XAGE-1b mRNA-negative specimens expressed XAGE-1 protein. Seropositivity was observed in 5 of 56 patients with adenocarcinoma, whereas none of 18 patients with other histologic types produced XAGE-1b antibody., Conclusion: XAGE-1b is highly and strongly expressed in lung adenocarcinoma and immunogenic in patients, suggesting that XAGE-1b is a promising antigen for immunotherapy against lung adenocarcinoma.

The morphogenesis of fungiform papillae occurs in a stereotyped pattern on the dorsal surface of the mammalian tongue via epithelial-mesenchymal interactions. These interactions are thought to be achieved via intercellular communication. Gap junctions can be observed in many developing tissues and have been suggested to participate in a variety of functions, including the regulation of cell proliferation, differentiation, and apoptosis. Here, we demonstrate that the expression of Connexin 43 (Cx43), a gap junction protein, is correlated significantly with the development of fungiform papillae, which exhibit a pattern formation and morphogenesis similar to the development of other epithelial appendages. Antisense-oligodeoxynucleotide (AS-ODN) against Cx43 was used to assess the developmental functions of Cx43. The expression patterns of the signaling molecules were disrupted by Cx43 inhibition. Interestingly, the expression patterns of Shh, a key molecule in the determination of the spacing patterns of fungiform papillae, were disturbed after treatment with Cx43 AS-ODN. We have also attempted to determine the functions of Bmp-2 by applying NOGGIN protein to tongue cultures. Our results indicate that upstream regulation via Cx43 controls the Shh and Bmp-2 pathways for the morphogenesis and pattern formation of fungiform papillae.

Background: The purpose of this study was to elucidate why odontogenic keratocysts (OKC) can form cystic lesions but not tumor masses, notwithstanding their prominent proliferative activity., Methods: We investigated cellular proliferation, cell death, and expression of apoptosis-related proteins in the lining cells of OKCs and of dentigerous cysts (DGCs)., Results: TdT-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells were observed in the surface layers of OKCs and of DGCs. However, no TUNEL-positive cells were seen in the basal or intermediate layers of both cysts. Ki67-positive ratio in the intermediate layer was the highest in OKCs. The p53-positive ratio of the intermediate layer was highest in OKCs. Bcl-2-positive cells were discernible exclusively in the basal layer of OKCs., Conclusions: These results suggest that cellular proliferation and death is regulated in association with apoptosis-related proteins in the lining epithelia of OKCs, and subsequently those cysts are seen as cystic lesions but not as tumor masses.

The purpose of the current study was to evaluate the bone formation when beta-tricalcium phosphate (TCP) was implanted in bone defects of rat femurs. beta-TCP granules were applied to defects created in the femurs of 65 male rats who were sacrificed 3, 7, 10, 14 or 30 days later. Bone tissues were embedded in paraffin, serial sections were cut and then stained with hematoxylin-eosin. Histomorphometric analyses were also conducted. Furthermore, total mRNAs were extracted, homogenized, and reverse transcribed, after which quantitative PCR assays were conducted with a LightCycler using the double-stranded DNA dye Syber Green I with primers for either rat osteopontin or osteocalcin. Tissues in defects without beta-TCP were used as controls. The amount of newly formed bone tissue in the beta-TCP implanted group was significantly greater in both the side areas and the central area of defects than in the control group. Expressions of osteopontin and osteocalcin mRNAs of cells in the defects of the experimental group were up-regulated compared with the control group at all time periods. Taken together, these results prove that beta-TCP is an appropriate material for osteoconduction and promotes bone formation in bone defects.

Aquaporins (AQPs) are a family of channel proteins that allow water or very small solutes to pass, functioning in tissues where the rapid and regulated transport of fluid is necessary, such as the kidney, lung, and salivary glands. Aquaporin-5 (AQP5) has been demonstrated to localize on the luminal surface of the acinar cells of the salivary glands. In this paper, we investigated the expression and function of AQP5 in the secretory granules of the rat parotid gland. AQP5 was detected in the secretory granule membranes by immunoblot analysis. The immunoelectron microscopy experiments confirmed that AQP5 was to be found in the secretory granule membrane. Anti-AQP5 antibody evoked lysis of the secretory granules but anti-aquaporin-1 antibody did not and AQP1 was not detected in the secretory granule membranes by immunoblot analysis. When chloride ions were removed from the solution prepared for suspending secretory granules, the granule lysis induced by anti-AQP5 antibody was inhibited. Furthermore, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, an anion channel blocker, blocked the anti-AQP5 antibody-induced secretory granule lysis. These results suggest that AQP5 is, expressed in the parotid gland secretory granule membrane and is involved in osmoregulation in the secretory granules.

We examined expression and function of osteopontin (OPN) in oral cancer cell lines using antisense oligonucleotide (AS). Quantitative real-time RT-PCR showed that expression in BSC-OF cells was significantly higher (10-fold) than that in KB cell. AS-study showed that foci of AS-treated BSC-OF cells possessed thin processes and radiated morphologically, although BSC-OF cells showed round foci. Cell growth in AS-group was lower (<80%) than the control. Invasion ability in AS-group became significantly lower (P<0.01). These results suggest that BSC-OF cell is useful for over-expression of OPN, and that OPN contributes to morphology, growth and invasion.

Aim: To investigate the expression of connexin 43 (CX43) mRNA in young and old human dental pulp tissues to determine the characteristics of CX43 expression., Methodology: Samples were obtained from human dental pulp of healthy young (17-23 years) and aged (>50 years) subjects. CX43 expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between value in the young pulp and that in the aged pulp was statistically analysed., Results: RT-PCR analysis of human dental pulp tissue detected CX43 mRNA in all the samples. CX43 was abundantly expressed in young adult dental pulp, but expression of CX43 mRNA was dramatically decreased in aged human dental pulp. QRT-PCR analysis also showed the reduced expression of CX43 in aged pulp, and expression of CX43 in young pulp was significantly higher (about 10-fold, P < 0.01, Mann-Whitney U-test)., Conclusion: Reduction of CX43 expression may be associated with the loss of viability in human dental pulp, and is considered as one characteristic of aged pulp.

Screening tests for glutamate producing strains were carried out, with the media containing carbohydrate and ammonia source as chief ingredients. Glutamate as well as certain other amino acids was detected by paper chromatography in culture broth of many microorganisms tested. Accumulation of L-glutamate in a significant amount (at least a few mg of glutamate per ml of broth) has been demonstrated by various strains of bacteria, streptomycetes, yeasts, and fungi. The highest level of glutamate production has been obtained by a new species of Micrococcus, yielding as much as 0.25 mole of it from one mole of glucose. The courses of fermentations mainly by known strains of microorganisms are shown. The importance of the cultural condition and strain specificity for the production of amino acids are briefly described.

The purpose of this study was to investigate the behavior of rat bone marrow cells (RBM) growing on surfaces with different pore sizes. RBM behavior on Millipore filters (MF-Millipore membrane filter) made from cellulose mixed esters with 5 different pore surfaces (0.45 microm, 1.2 microm, 3.0 microm, 5.0 microm and 8.0 microm) were compared in terms of morphological changes on the different pore sizes. Furthermore, the expressions of osteopontin and osteocalcin mRNAs were investigated. On the 1.2 microm and 3.0 microm pore surfaces, RBM attached to the substrate well, but cells on the 5.0 microm and 8.0 microm pore surfaces invaded deeply into the pores. Higher levels of both osteopontin and osteocalcin mRNA expression were always observed in cells cultured on the 1.2 microm filter. These results suggest that the 1.2 microm Millipore filter pore size is the most suitable for inducing RBM to differentiate into an osteoblastic phenotype among these surfaces and is probably related to production of the ECM but not to the phenomenon of cell spreading.

This case report describes a rare case of canalicular adenoma arising in the upper lip of a 61-year-old male patient. Macroscopic examination of the tumor revealed a well-defined, smooth, firm, elastic hard, round nodule with a diameter of 1.0 cm. The cut surface was white. Histopathology showed that the tumor was an encapsulated mass with a complex cellular pattern of anastomosing duct-like or trabecular structures lined by a single layer of tall columnar epithelial cells, which were embedded in a loose, fibrous, and highly vascular connective tissue stroma. The tumor cells were immunoreactive to AE1/AE3, CK19 and S-100, were partially positive for CK7, CK8, GFAP and PCNA, but were negative for SMA, CK13, CK14 and vimentin.

Case Report: The authors present a case of a patient with an abdominal CSF pseudocyst that resulted from an allergic reaction to silicone. The patient underwent repair surgery of the meningomyelocele associated with the Chiari II malformation, and the V-P shunt was instituted at 6 months of age. A formation of the abdominal CSF pseudocyst and the consequent shunt malfunction were observed 40 days after the V-P shunt. An increase in the number of the peripheral eosinophils and serum immunoglobulin E (IgE), and an infiltration of eosinophils in the specimen harvested from the pseudocyst wall suggested an allergic reaction as the cause of the pseudocyst. A sixth operation to revise the V-P shunt was performed using the shunt system made of "extracted silicone", which was produced extracting the allergic substances., Outcome: The serum IgE was normalized after surgery and the abdominal CSF pseudocyst has not recurred for 22 months.

Background and Objective: It is still an open question why long junctional epithelium can proliferate and occupies the root surface following periodontal surgery or experimentally produced periodontitis, and why the epithelium repopulated once on the root surface is replaced by the connective tissue. The aim of this study is to investigate the proliferative activity of the newly formed regenerative connective tissue and long junctional epithelium during wound healing by staining argyrophilic proteins of the nucleolar organizer regions (AgNORs)., Methods: Regenerative connective tissue and long junctional epithelium were experimentally created by insertion of a rubber piece between maxillary molars of rats for 1 week. After removal of the rubber, AgNORs parameters including nuclear area (NA), AgNORs area (AA), AgNORs percentage nuclear area (APNA), AgNORs number (AN) and nuclear number (NN) in regenerative connective tissue and long junctional epithelium were measured and analyzed statistically., Results: APNA in long junctional epithelium after 1 and 4 weeks was over two times greater than that in the regenerative connective tissue. AA in long junctional epithelium was significantly higher than in regenerative connective tissue at 1 and at 4 weeks post-treatment. AN was higher in the central portion than at the root surface except at 20 weeks. APNA and AA decreased remarkably in long junctional epithelium at 12 weeks post-treatment (approximately half at 4 weeks), whereas in regenerative connective tissue, they did not change distinctly., Conclusions: These results imply that long junctional epithelium cannot supply sufficient epithelial cells because of their significantly low rates of proliferation, consequently long junctional epithelium becomes shorter after 12 weeks, whereas the proliferative activity of regenerative connective tissue maintains the same level of proliferation, and ultimately long junctional epithelium is replaced by regenerative connective tissue.

This case report describes an acinic cell carcinoma found by a recurrence of a ranula in the sublingual gland. A 42-year-old male was admitted to the hospital of the Tokyo Dental College with a swelling in his right oral floor but without pain. The lesion was treated by windowing the same day under the diagnosis of a ranula, but the swelling appeared again at the same area eight months after the first operation. A resection was performed, and the specimen was sent to the clinical laboratory for pathological diagnosis. Proliferating serous cells were seen in part of the wall of an exudative mucous cyst. PAS staining was partially positive, and immunohistochemical staining for S-100 protein, lactoferrin, and amylase were also positive in cytoplasmic granules. This report concludes that the pathological diagnosis is beneficial in clarifying the reasons for the recurrence of a benign lesion.

By serologic identification of antigens by recombinant expression cloning (SEREX) analysis using an autologous lung adenocarcinoma cell line, OU-LU-6, as a cDNA library source, we demonstrated that XAGE-1 was the dominant antigen recognized by serum from a patient. By immunoscreening, we obtained 38 positive cDNA clones consisting of 16 genes designated as OY-LC-1 to -OY-LC-16. OY-LC-1, represented by 18 clones, was identical to XAGE-1. OY-LC-2 to -16, represented by either a single or 2 clones, were identical to known genes shown to be ubiquitously expressed in various normal tissues. RT-PCR analysis showed that of 4 XAGE-1 transcripts-XAGE-1a, b, c and d-XAGE-1b was expressed in OU-LU-6 dominantly. Furthermore, XAGE-1b mRNA was expressed in 4 of 10 lung cancer tissues, whereas no expression was observed in normal tissues. Of 4 XAGE-1b mRNA positive cancer tissues, 3 were adenocarcinoma and one was poorly differentiated squamous cell carcinoma. Of 32 sera from lung cancer patients, 8 sera were reactive with the XAGE-1b product. Those 8 sera were from patients with adenocarcinoma. These findings indicated strong immunogenicity of XAGE-1b in lung adenocarcinoma and suggested its potential use as a target for vaccine-based immunotherapies., (Copyright 2003 Wiley-Liss, Inc.)

To investigate the time sequence of the neural processes underlying face perception, magnetoencephalography was performed using a seeing-as-face task, in which visual inputs were identical across two conditions, but subject perceptions differed: one being a non-specific pattern of geographical shapes, the other being a percept of a face. Subtraction between the two conditions revealed a response occurring 120 ms after stimulus onset in right occipital, approximately 50 ms earlier than previously reported response at a latency of 170 ms at the right fusiform gyrus. As our novel task completely excluded differences in low-level properties of visual stimuli between control and face conditions, these two responses were considered specific to face perception. The result supported the two-stage theory of face processing.