Large scale replication analysis of loci associated with lipid concentrations in a Japanese population.

BACKGROUND: Recent genome wide association studies discovered seven novel loci that influence plasma concentrations of triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol in Europeans. To date, large scale replication studies using populations with known differences in genome-wide linkage disequilibrium (LD) pattern have not been undertaken. METHODS: To address this issue, we tested associations between single nucleotide polymorphisms (SNPs) within the seven novel loci and plasma lipid profiles in 21 010 Japanese individuals. RESULTS: Multiple linear regression analyses showed that the rs3812316 in MLXIPL was strongly associated with triglyceride concentrations (p approximately 3.0x10(-11), 7.1 mg/dl decrease per minor C allele) and that rs599839 in CELSR2/PSRC1/SORT1 was strongly associated with LDL cholesterol concentrations (p approximately 3.1x10(-11), 4.7 mg/dl decrease per minor G allele) in the Japanese population. SNPs near ANGPTL3, TRIB1 and GALNT2 showed evidence for associations with triglyceride concentrations (3.6x10(-6)

Gene expression profiles in Ciona intestinalis tailbud embryos.

A set of 3423 expressed sequence tags derived from the Ciona intestinalis tailbud embryos was categorized into 1213 independent clusters. When compared with DNA Data Bank of Japan database, 502 clusters of them showed significant matches to reported proteins with distinct function, whereas 184 lacked sufficient information to be categorized (including reported proteins with undefined function) and 527 had no significant similarities to known proteins. Sequence similarity analyses of the 502 clusters in relation to the biosynthetic function, as well as the structure of the message population at this stage, demonstrated that 390 of them were associated with functions that many kinds of cells use, 85 with cell-cell communication and 27 with transcription factors and other gene regulatory proteins. All of the 1213 clusters were subjected to whole-mount in situ hybridization to analyze the gene expression profiles at this stage. A total of 387 clusters showed expression specific to a certain tissue or organ; 149 showed epidermis-specific expression; 34 were specific to the nervous system; 29 to endoderm; 112 to mesenchyme; 32 to notochord; and 31 to muscle. Many genes were also specifically expressed in multiple tissues. The study also highlighted characteristic gene expression profiles dependent on the tissues. In addition, several genes showed intriguing expression patterns that have not been reported previously; for example, four genes were expressed specifically in the nerve cord cells and one gene was expressed only in the posterior part of muscle cells. This study provides molecular markers for each of the tissues and/or organs that constitutes the Ciona tailbud embryo. The sequence information will also be used for further genome scientific approach to explore molecular mechanisms involved in the formation of one of the most primitive chordate body plans.

Role of hydrogen peroxide in antitumor activity induction by sodium 5,6-benzylidene-L-ascorbate.

Role of hydrogen peroxide (H2O2) in the induction of antitumor activity against chemically-induced rat hepatocellular carcinoma by sodium 5,6-benzylidene-L-ascorbate (SBA) was investigated. ESR spectroscopy demonstrated that rat liver homogenate of cancerous tissue significantly enhanced the radical intensity of SBA more potently than that of precancerous or normal tissue. The peroxyoxalate chemiluminescence method demonstrated that SBA significantly enhanced the production of H2O2-derived chemiluminescence intensity in the liver homogenates, and the effect of SBA was greater in cancerous tissue than in precancerous or normal tissue. Addition of ascorbic acid, a degradation product of SBA, showed similar but slightly weaker stimulation effects. These data suggest that antitumor activity of SBA in vivo might, at least in part, be due to H2O2 production.

Ameloblastic fibrosarcoma arising de novo in the maxilla.

An ameloblastic fibrosarcoma (AFS) arising in the maxilla of a 14-year-old male is described. The tumor originated from the alveolar bone of the right maxilla with no apparent history of pre-existing lesion. Histologically, the lesion was composed of benign-appearing epithelial islands and strands scattered within an exceedingly cellular mass of mesenchymal tissue comprising a large number of stellate-and spindle-shaped fibroblast-like cells with marked pleomorphism. Occasional cementum-like calcification was also noted. Immunohistochemically, the neoplastic mesenchymal cells were positive only for vimentin, whereas the ameloblast-like epithelial component showed a distinctly positive reaction for wide-spectrum keratin and squamous cytokeratin. Clinicopathological features of the current case, as well as previously reported examples of AFS originating from the maxilla, are briefly discussed.

DNA-based diagnostics in the study of heritable and acquired disorders.

Recent advances in molecular biology techniques have made DNA diagnostics a reality. A major advance was the development of the polymerase chain reaction, which allows the direct analysis of a region of a known gene from minute quantities of template DNA. The DNA can even be isolated from such sources as saliva, hair roots, microscope slides, paraffin-embedded tissue, and clinical swabs, in addition to blood and biopsy material. Another major improvement is the development of automated procedures suitable for robotic stations. Together these improvements make DNA analysis an important part of diagnostic procedures in studying the etiology of diseases.

Immunohistochemical analysis of EGF in epiphyseal growth plate from normal, hypophysectomized, and growth hormone-treated hypophysectomized rats.

Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.

Computer-aided image analysis applied to immunogold-silver staining: evaluation of proliferating cell nuclear antigen (PCNA)-reactive sites in paraffin sections.

Feasibility of the combination of the immunogold-silver staining method (IGSS) and computer-aided image analysis was assessed for the detection of antigen in an immunostained, paraffin-embedded section. Using low-temperature IGSS, we stained a specimen of human oral squamous cell carcinoma with a monoclonal antibody, PC 10, against a proliferating cell nuclear antigen (PCNA/cyclin), and the section was analyzed by ACAS 570 interactive laser cytometry. The PCNA-positive cells, exhibiting a heteromorphic texture, were contrasted by the dark staining of their nuclei, but showed heterogeneity in staining intensity from cell to cell. Using a conventional microscope light source rather than a laser, and by employing the COMPLEMENT DATA program (which permits inversion of the data values) installed in the ACAS 570 software system, we were able to obtain a 'complemental image' which replicated the real immunohisto-morphology. Approximately 30-35 cells from three different areas in the same section were selected by DEFINE CELL and MARK AREA programs, and quantitative image analysis was performed in terms of cell integrated value, area, perimeter, and shape factor indicated in histogram form. The combined utilization of IGSS with computer-aided image analysis was demonstrated to offer a crucial advantage for the quantitative assessment of immunostained sections.

Increased expression of type VI collagen genes in drug-induced gingival enlargement.

Fibrotic gingival enlargements induced by phenytoin or nifedipine were examined with special reference to type VI collagen expression. Immunolocalization studies showed abnormal accumulation of type VI collagen around the collagen fiber bundles in the fibrotic gingival enlargements. Examination of total RNA extracted from fibroblasts and tissues of enlarged gingivae demonstrated increased type VI collagen steady-state mRNA levels. These results suggest that excessive deposition of type VI collagen in drug-induced gingival enlargement is attributed to increased expression of the collagen genes.

Epidermoid carcinoma originating from the gingival sulcus.

This paper describes a case of epidermoid carcinoma originating from the gingival sulcular epithelium with proper plane of histopathology. Relevant articles on carcinoma of gingiva are briefly reviewed, and the necessity for early diagnosis of this lesion is emphasised.

Immunohistochemical detection of sialyl Le(x) antigen on mucosal Langerhans cells of human oral mucosa following neuraminidase pretreatment.

Histochemical assessment of selected carbohydrate sequences on Langerhans cells of human oral mucosa was made by combined use of enzyme digestion and immunostaining with monoclonal antibodies against specific carbohydrate structures. In both frozen sections and epithelial sheets without the enzyme pretreatment, mucosal Langerhans cells, identified by positive staining with anti-CD1a and HLA-DR antibodies, did not express any carbohydrate antigens on their surface. In contrast, following neuraminidase pretreatment of both types of material, the fucosylated type 2 chain (Le(x)) became detectable on Langerhans cells, indicating that sialic acid is the terminal residue of this sequence. Other enzymes were ineffective in this apparent unmasking, and the staining patterns of the other related carbohydrate sequences (Le(y)+, Le(a), Le(b)) remained unaffected by pretreatment with any of the enzymes used. These findings suggest that the mucosal Langerhans cells possess a unique carbohydrate chain, the sialyl fucosylated type 2 sequence (sialyl Le(x) antigen).

Immunohistochemical demonstration of epidermal growth factor in chondrocytes of mouse femur epiphyseal plate.

The distribution of epidermal growth factor (EGF) has been studied immunohistochemically in chondrocytes of femur epiphyseal plate cartilage from adult mice. The chondrocytes of the resting and proliferating zones reacted positively, while those of maturing and calcifying zones showed a relatively weak reaction. The immunoreaction was observed over both the cytoplasm and nuclei of resting and proliferating chondrocytes and was prominent in the proliferating chondrocytes. These findings, taken together with the suggested synergistic action of EGF with other growth factors, raise the possibility that chondrocytes, particularly in the proliferating zone, participate in a regulatory mechanism by EGF on cell division and growth in the epiphyseal plate cartilage.

Ameloblastoma arising in calcifying odontogenic cyst. Report of a case.

An unusual case of an ameloblastoma with adenoid-cribriform features arising in calcifying odontogenic cyst is reported. A possible relationship between this peculiar variant of calcifying odontogenic cyst and a conventional ameloblastoma is discussed.

Cylindrical cell papilloma of the nasal cavity and paranasal sinuses. A histochemical and cytochemical study.

A cylindrical cell papilloma occurring in the sino-nasal mucosa of a 61-year-old woman was studied histochemically at both light and electron microscopic levels. The cylindrical cells demonstrated distended intracytoplasmic microcysts with numerous microvillous projections on the apical cell membrane. The retained mucosubstance in the cystic spaces stained intensely with HID-TCH-SP sequence and showed no apparent communication with the extracellular space. These findings indicate that the cylindrical cells may be of mucous-secreting cell derivation, being derived from cells possessing abnormal and/or failed secretory function. The property of sulfated mucin in the 'cylindric' intracytoplasmic cysts is suggested to be the result of endodermal displacement or metaplasia of the Schneiderian epithelium.

Increased gene expression of matrix metalloproteinase-3 (stromelysin) in skin fibroblasts from patients with severe recessive dystrophic epidermolysis bullosa.

Gene expression of matrix metalloproteinase 3 (MMP-3 = stromelysin) was examined in the skin fibroblasts obtained from patients with severe recessive dystrophic epidermolysis bullosa (RDEB). Steady-state mRNA level of MMP-3 was selectively increased in the unstimulated RDEB cells by a post-transcriptional mechanism. A parallel study on the susceptibility of type VII collagen to MMPs revealed that this type of collagen is degraded by MMP-3, but not by MMP-1 (collagenase). These data suggest that MMP-3 may play an important role in the blister formation fo the skin in RDEB patients by the degradation of anchoring fibrils consisting of type VII collagen.

Immunocytochemical detection of S-100 protein in rat mandibular condylar cartilage.

Light and electron microscopic immunocytochemistry was used to localize S-100 protein. It was demonstrated in chondrocytes of the proliferative and hypertrophic zones but none could be found in the fibrous articular layer. The staining intensity for S-100 protein was stronger in the hypertrophic cells than in chondrocytes from the proliferative zone. Ultrastructurally, immunoreactive gold particles were detected in the cytosolic region, closely apposed to the profiles of endoplasmic reticulum with occasional dense aggregations. A few gold particles were seen on the nuclear chromatin and on condensing vacuoles of Golgi complexes. Because of the calcium-binding and possibly fatty acid-binding properties of the protein molecule, the immunocytochemical expression of S-100 protein in the chondrocytes may be involved in cartilage cell differentiation, metabolism and mineralization.

Light and electron microscopic demonstration of wheat germ agglutinin binding in the pericellular matrix of rat mandibular condylar cartilage.

Binding of wheat germ agglutinin to rat mandibular condylar cartilage was investigated with the avidin-biotin peroxidase complex. Binding sites were observed in the pericellular matrix of the hypertrophic cell zone. Two distinct patterns were identified: one showed binding to the pericellular matrix without apparent contour; in the other binding was confined to the matrix but with conspicuous condensation forming a pericellular rim. The first binding pattern was seen particularly in the upper part of the hypertrophic cell zone adjoining the mature cell zone; the second was localized in the lower part of this zone adjacent to the site of endochondral calcification. The binding sites in the pericellular matrix are assumed to be NANA and/or GlcNAc in view of the sugar specificity of this lectin. The presence of these binding sites in this region may be due to a structural alteration or modification of proteoglycans in the course of preparation for endochondral calcification.

An ameloblastoma-like neoplasm of the buccal mucosa. Report of a case.

A case of an unusual neoplasm of the buccal mucosa nearly identical to an ameloblastoma is reported. The tumor probably arose from the overlying squamous epithelium in the vicinity of the parotid duct.

[Report on clinico-pathological examinations in Meikai University (formerly Josai Dental University) Hospital (4)].

This clinico-pathological study was conducted to examine biopsies and surgical specimens for histopathological diagnosis from April 1983 to March 1988 in Meikai University (formerly Josai Dental University) Hospital. The following statistical results were obtained: 1) The total number of the specimens examined was 2705, 2026 from the dental clinic and 679 from the medical clinic. 2) Concerning age distribution, most specimens from the dental clinic were from patients in the 40's age bracket, whereas most from the medical clinic were from those under 10 years of age. With regard to sex distribution, the difference between males and females was not so much in the dental clinic specimens, but the number from the medical clinic was greater for males than for females. 3) Concerning classification of the tumors from the dental clinic, 181 cases were benign; and of them 41 were odontogenic and 140 were non-odontogenic tumors. Among the odontogenic tumors, ameloblastoma, cementoma, and odontoma were the most numerous. Among non-odontogenic tumors, fibroma was the most common, followed by pleomorphic adenoma. Malignant tumors comprised 77 cases. Of them squamous cell carcinoma was the most commonly found, with the grade-2 type being most numerous. Of the medical clinic tumors, nevus was the most common benign tumor observed; and adenocarcinoma, the most common malignant one. 4) Concerning the distribution of main lesions in the dental clinic specimens, most specimens were cysts or cystic lesions, followed by inflammatory lesions and tumors. Among medical clinic specimens, inflammatory lesions were the most common. 5) Regarding the dental clinic specimens, the most frequently observed case was the radicular cyst (301 samples) among the odontogenic cysts (514 samples); and among non-odontogenic cysts (290 samples), the most common was the mucous cyst (159 samples). 5) In terms of the distribution by location, tissue or organ, most tumors from the dental clinic were from the jaws, followed in descending frequency by those from gingiva and alveolar mucosa, maxillary sinus, lip, buccal mucosa, and tongue. Among the medical clinic tumors, the paranasal cavity and tonsils were the most frequent locations.