Insulin-like growth factor-binding protein-2 and -3 in gingival crevicular fluid.

BACKGROUND AND OBJECTIVE: Insulin-like growth factor-binding proteins (IGFBPs) are crucial regulators of insulin-like growth factor (IGF). They enhance or inhibit IGF functions, but also exhibit IGF-independent effects. In a previous study, we detected, qualitatively, IGFBP-2 and -3 in gingival crevicular fluid using a cytokine antibody array. Here we extended these results using an ELISA to determine the concentrations of IGFBP-2 and -3 in gingival crevicular fluid. In addition, we explored whether the expression of IGFBP-2 and IGFBP-3 correlates with periodontal disease severity. MATERIAL AND METHODS: Gingival crevicular fluid samples from 92 sites of 12 patients affected with periodontal disease and from 100 sites of 19 healthy volunteers, were collected, divided into two groups and analyzed by ELISA for IGFBP-2 and -3 expression. The potential correlation among probing depth, gingival index and the concentrations of IGFBP-2 and -3 was analyzed. RESULTS: Positive correlations were observed between the concentration of IGFBP-2 and probing depth and gingival index, but not for IGFBP-3. The IGFBP-2 concentrations at bleeding on probing-positive sites and at sites with a probing depth of ≥ 4 mm were higher than at bleeding on probing-negative sites and at sites with a probing depth of ≤ 3 mm. CONCLUSION: These results indicate that IGFBP-2 is a potential novel marker for periodontal disease progression. As IGFBP-2 modulates bone metabolism and cell migration, IGFBP-2 in the gingival crevicular fluid may reflect periodontal disease activity.

TGF-ß signaling in gingival fibroblast-epithelial interaction.

The underlying mechanism and the therapeutic regimen for the transition of reversible gingivitis to irreversible periodontitis are unclear. Since transforming growth factor (TGF)-ß has been implicated in differentially regulated gene expression in gingival fibroblasts, we hypothesized that TGF-ß signaling is activated in periodontitis-affected gingiva, along with enhanced collagen degradation, that is reversed by TGF-ß inhibition. A novel three-dimensional (3D) gel-culture system consisting of primary human gingival fibroblasts (GF) and gingival epithelial (GE) cells in collagen gels was applied. GF populations from patients with severe periodontitis degraded collagen gels, which was reduced by TGF-ß-receptor kinase inhibition. Up-regulation of TGF-ß-responsive genes was evident in GF/GE co-cultures. Furthermore, the TGF-ß downstream transducer Smad3C was highly phosphorylated in periodontitis-affected gingiva and 3D cultures. These results imply that TGF-ß signaling is involved in fibroblast-epithelial cell interaction in periodontitis, and suggest that the 3D culture system is a useful in vitro model for therapeutic drug screening for periodontitis.

p73 is highly expressed in myoepithelial cells and in carcinomas with metaplasia.

p73, a homologue of p53 gene, is expressed in several normal tissues including central nervous system, but data regarding tumors are scant. In this study, we have analyzed the status and expression of the p73 gene in primary breast tumors, as well as 4 normal salivary glands, 2 carcinomas with metaplasia and mixed tumors. We found that periductal myoepithelial cells of all of the mammary gland examined were clearly stained with the specific anti-p73 antibody. Furthermore, we found the expression of p73 in the neoplastic myoepithelial cells in carcinomas with metaplasia and in mixed tumors. The findings in the present study provide valuable information on the characteristics of myoepithelial cells.

Complete primary structure of rainbow trout type I collagen consisting of alpha1(I)alpha2(I)alpha3(I) heterotrimers.

The subunit compositions of skin and muscle type I collagens from rainbow trout were found to be alpha1(I)alpha2(I)alpha3(I) and [alpha1(I)](2)alpha2(I), respectively. The occurrence of alpha3(I) has been observed only for bonyfish. The skin collagen exhibited more susceptibility to both heat denaturation and MMP-13 digestion than the muscle counterpart; the former had a lower denaturation temperature by about 0.5 degrees C than the latter. The lower stability of skin collagen, however, is not due to the low levels of imino acids because the contents of Pro and Hyp were almost constant in both collagens. On the other hand, some cDNAs coding for the N-terminal and/or a part of triple-helical domains of proalpha(I) chains were cloned from the cDNA library of rainbow trout fibroblasts. These cDNAs together with the previously cloned collagen cDNAs gave information about the complete primary structure of type I procollagen. The main triple-helical domain of each proalpha(I) chain had 338 uninterrupted Gly-X-Y triplets consisting of 1014 amino acids and was unique in its high content of Gly-Gly doublets. In particular, the bonyfish-specific alpha(I) chain, proalpha3(I) was characterized by the small number of Gly-Pro-Pro triplets, 19, and the large number of Gly-Gly doublets, 38, in the triple-helical domain, compared to 23 and 22, respectively, for proalpha1(I). The small number of Gly-Pro-Pro and the large number of Gly-Gly in proalpha3(I) was assumed to partially loosen the triple-helical structure of skin collagen, leading to the lower stability of skin collagen mentioned above. Finally, phylogenetic analyses revealed that proalpha3(I) had diverged from proalpha1(I). This study is the first report of the complete primary structure of fish type I procollagen.

Isolation and characterization of a novel human gene (NESH) which encodes a putative signaling molecule similar to e3B1 protein.

Using a conventional cloning technique, a novel full-length cDNA was isolated and sequenced from a human placental cDNA library. This cDNA consists of 2129 bp and has a predicted open reading frame encoding 366 amino acids. It possesses a Src homology 3 (SH3) motif, proline-rich region, serine-rich region and no catalytic domain, suggesting that it seems to be a signaling protein most similar to e3B1, an eps8 SH3 binding protein. PCR-based mapping with both a monochromosomal hybrid panel and radiation hybrid cell panels placed the gene to human chromosome 17q21.3 near the marker D17S1795.

Resection of a hilar cholangiocarcinoma in a patient with absent portal bifurcation.

Absence of the bifurcation of the portal vein is an extremely rare anomaly and the misled ligation of the portal vein would result in the lethal hepatic failure. In this paper, a resected case of hilar cholangiocarcinoma with this anomalous portal venous system is first presented. Preoperative percutaneous transhepatic portography disclosed an absence of portal bifurcation and a transversely running portal vessel from the right anterior segment to the left lateral segment. The patient underwent left hepatic lobectomy with caudate lobectomy, in which the portal trunk was successfully preserved.

Molecular cloning and characterization of a novel human gene (NESCA) which encodes a putative adapter protein containing SH3.

A full-length cDNA encoding a novel protein was isolated and sequenced from a human placental cDNA library. This cDNA consists of 1990 bp and has a predicted open reading frame encoding 433 amino acids. It possesses an Src homology 3 (SH3) motif, a leucine zipper motif and no catalytic domain, suggesting that it seems to be an adapter protein. PCR-based mapping with both a monochromosomal hybrid panel and radiation hybrid cell panels placed the gene to human chromosome 1q21-22.

Critical amino acid substitutions in the Src SH3 domain that convert c-Src to be oncogenic.

The Src homology 3 (SH3) domain, originally identified in v-Crk, plays an important role in signal transduction. The comparative study with c-src has revealed that v-src oncogene of Schmidt-Ruppin strain of Rous sarcoma virus has three point mutations in its SH3 domain and one in the upstream of SH3. To assess the role of these mutations, each of the single mutations was introduced into c-Src by oligonucleotide-directed mutagenesis and its effect on cell transformation was examined. While variant Src proteins that carry each one of single mutations could not transform cells, double mutation at positions 95 and 117 converted c-Src to be oncogenic and active in kinase. An additional mutation at position 124 together with one at 95 and 117 further activated Src kinase. By use of GST-fusion forms of v-Src SH3 and c-Src SH3, we found that these mutations in SH3 suppressed the binding of SH3 with c-Src protein, possibly with a linker region, while v-SrcSH3 retained the ability to bind a subset of cellular protein to the level similar to those of c-SrcSH3. Taken together, our results suggest that point mutations accumulated in SH3 region can activate, in concert, Src kinase by relaxing the interaction between SH3 and the linker region and subsequently convert Src to be oncogenic.

Report of a case of benign cystic mesothelioma.

A 32-yr-old man was admitted to Anjo Kosei Hospital for the examination of a cystic peritoneal lesion. This patient had a past medical history significant for an appendectomy. Ultrasonography and computed tomography revealed multiple cysts in the abdominal and pelvic cavities. An exploratory laparotomy was performed on July 1, 1993, revealing numerous thin-walled and translucent cysts in the serosal tissues of the parietal and visceral abdominal peritoneum. These cysts were excised surgically because of clinical suspicion of pseudo-myxoma peritonei. Microscopic examination revealed cysts within the thin cyst wall that were composed of fibrous connective tissue and lined by a single layer of cuboidal or flattened epithelium. Immunohistochemical and ultrastructural studies demonstrated that the lining cells were of mesothelial origin, and the diagnosis was benign cystic mesothelioma. In this paper, we present appropriate diagnostic and treatment procedures for benign cystic mesothelioma and discuss the particular usefulness of using immunohistochemical methods to achieve a histological diagnosis.

Release of lipoteichoic acid from Staphylococcus aureus by treatment with cefmetazole and other beta-lactam antibiotics.

The effect of cefmetazole on the growth together with the release of cellular lipoteichoic acid from cefazolin-resistant strains of Staphylococcus aureus was compared with that of cefazolin, cefotiam, cefoxitin and cefuroxime. Bacteriolytic actions were measured by turbidity and bactericidal actions were followed by viable cell count. Release of cellular lipoteichoic acid was measured by the radioactivity in the supernatant of the cultures. Cefmetazole exerted more potent effects on the bacterial growth and induced more marked release of cellular lipoteichoic acid from resistant strains as compared with other beta-lactams.

Purification and properties of chromosomally mediated beta-lactamase from Citrobacter freundii GN7391.

Both a penicillinase and a cephalosporinase are present in a strain of Citrobacter freundii (GN7391) resistant to beta-lactam antibiotics. The penicillinase was identical to the type Ia penicillinases (Type III by Richmond classification), mediated by Rms212 and R-TEM. A cephalosporinase, typical of enterobacteriaceae chromosomal beta-lactamase (Type I by Richmond classification), was purified from the strain. It gave a single protein band on polyacrylamide gel electrophoresis and immunoelectrophoresis; the pI was 8.6 and its molecular weight was approximately 38 000. Cysteine was not found among its amino acids. The specific activity was 388 units (mg protein)-1 for the hydrolysis of cephaloridine, and the optimal pH was 8.0. Rabbit antiserum obtained against the purified enzyme showed cross-reaction with cephalosporinases produced by strains of Enterobacter cloacae in a neutralization test.

An autopsy case of atypical senile dementia with atrophy of the temporal lobes--a clinical and histopathological report.

A man aged 70, descendant of an apparently healthy family, showed disorientation, delusional ideas and rages at 66. Later there was slowly advancing deterioration with muteness, disorientation and dysphagia. He died of cardiac failure. There was the diffuse atrophy of the cerebrum, which was remarkably accentuated on both temporal lobes (poles, T2 and T3), where the loss of nerve cells and proliferation of astrocytes were found in the cortex and pallor and conspicuous fibrillary gliosis were noted in the white matter. These findings fundamentally suggest Pick's disease. On the one hand, numerous senile plaques and Alzheimer's neurofibrillary changes, suggestive of Alzheimer's disease (senile dementia), were observed throughout the cerebral cortex. On the other hand, a few inflated cells were also seen in the cingulate, superior frontal gyri and temporal lobes. Basilar artery was moderately atherosclerotic and cerebrovascular disorders were distributed throughout the cerebral cortex and basal ganglia, especially in the field of supply of middle cerebral artery. This case is similar to rare cases reported by Berlin (1949), Neumann (1949) and Oyanagi et al. (1975). The nosological situation as a disease entity remains to be determined.