A novel heterophilic antibody interaction involves IgG4.

IgG4 has been implicated in a diverse set of complex pathologies - e.g. autoimmune pancreatitis (AIP), idiopathic membranous nephropathy - and carries unique features including lack of activation of the classical complement pathway and a dynamic Fab-arm exchange. We recently showed that the rheumatoid factor (RF)-like activity of IgG4 is achieved through a hitherto unknown, Fc-Fc (and not Fab-Fc as is the case in classical RF; CRF) interaction; hence the name, novel RF (NRF). Here, we further explore the resemblance/difference between CRF and NRF. As heterophilic interactions of human IgM RF (CRF) are well known, we checked whether this is the case for IgG4. Human IgG4 showed variable reactivity to animal IgGs: reacting intensely with rabbit and mouse IgGs, but weakly with others. The binding to rabbit IgG was not through the Fab (as in CRF) but via the Fc piece, as was recently shown for human IgG (NRF). This binding correlates with the IgG4 concentration per se and could therefore be of diagnostic usage and incidentally explain some observed interferences in biological assays. In conclusion, here is defined a novel heterophilic antibody interaction and is established the universality of the unique Fc-Fc binding, both involving IgG4.

Local circular polarization observed in surface vortices of optical near-fields.

We demonstrate local circular polarization in surface vortices of an optical near-field generated by superposing two cross-propagating evanescent waves with transverse electric fields on a planar dielectric surface. The circularly rotating local electric fields are converted into circularly polarized propagating light waves in free space via a near-field interaction with a sub-wavelength size local probe. The results show that optical near-fields generated under the influence of a material environment with local rotational symmetry carry angular pseudo-momentum with respect to the symmetry axis. The local circular polarization is of fundamental significance in spin-related and magneto-optical phenomena in nanophotonics.

3D morphological assessment of occlusal treatment by measuring dental casts with a micro-focus X-ray CT.

The analysis of occlusal relationship is important for the success of dental treatment. Three-dimensional (3D) computer models of upper and lower dental casts can play a significant role. In this study, we proposed and applied a new method in actual clinical assessment to measure dental casts with occlusal relationship by using a micro-focus X-ray CT system. We examined the modelling accuracy by comparing multiple 3D images taken by shifting the dental cast position. Modelling accuracy was confirmed as 0.03 mm. One occlusal treatment in clinical practice was selected as a case example. The dental casts and bite impression, taken before treatment, were scanned and the occlusal contacts and distance distribution between the upper and lower casts were visualized by a coloured map and overlaid on the computer models. Distances between the upper and lower casts of selected points were compared before and after the treatment. Initially, the subject had early contact on the anterior teeth, where distance was measured as 0.04 mm, and only one area measured less than 0.15 mm. After treatment, five areas measured less than 0.15 mm. Also, by comparing the dental cast models taken before and after occlusal adjustment of the tooth, the position and amount of adjustment were visualized. We successfully demonstrated the quantitative clinical assessment of occlusal treatment.

Ingestion of paddy rice increases intestinal mucin secretion and goblet cell number and prevents dextran sodium sulfate-induced intestinal barrier defect in chickens.

Paddy rice is a potential feed grain for chickens, whose strong gizzards can crush the hull. Here, we investigated whether paddy rice rich in hull-derived water-insoluble dietary fiber stimulates intestinal mucin secretion and production, as well as the possible involvement of paddy rice in intestinal barrier function. Layer male chicks at 7 d of age were divided into four groups according to the diet: corn, polished rice, brown rice, or paddy rice (650 g/kg diet), which they ate for 14 consecutive days. At 21 d of age, the birds were refed their experimental diets, and small intestinal mucin fractions were collected to determine intestinal mucin content. Small intestinal mucin secretion was induced most strongly in the paddy rice group (Experiment 1). The rank order of diet-induced mucin secretion was paddy rice > corn = brown rice > polished rice. Ileal MUC2 gene expression and ileal number of goblet cells were highest in the paddy rice group (Experiment 1). A study of bromodeoxy-U uptake into ileal epithelial cells indicated the increase in goblet cells in the paddy rice group was related to accelerate epithelial cell migration (Experiment 2). A single supplementation of isolated rice hulls without kernels increased MUC2 gene expression and goblet cell numbers (Experiment 3), suggesting the importance of the hull's bulk-forming capacity on mucin production. Finally, chicks fed corn or paddy rice were orally administered dextran sodium sulfate (DSS) to disrupt intestinal barrier function. In the DSS-treated birds, the intestinal permeability of fluorescein isothiocyanate dextran in the everted gut sacs was much lower in the paddy rice group than in the corn group (Experiment 4), showing that paddy rice protects against mucosal disruption. In conclusion, ingestion of paddy rice increases intestinal mucin secretion and production through enhanced MUC2 gene expression and epithelial turnover and prevents DSS-induced intestinal barrier defects in chickens.

Comparison of Swedish interactive threshold algorithm and full threshold algorithm for glaucomatous visual field loss.

PURPOSE: To compare the prevalence of visual field loss, the sensitivity distribution, and the size and depth of glaucomatous visual field defects using the standard full threshold (FT) and the Swedish interactive threshold algorithm (SITA) standard (SS) procedures in patients with early or suspected glaucoma. METHODS: Automated perimetry findings were retrospectively evaluated in 53 patients (105 eyes) with early or suspected glaucoma. RESULTS: The number of eyes judged to have glaucomatous visual field loss by SS (48 eyes) was significantly larger than what was found with FT (35 eyes), and 70 eyes were classified as pre-perimetric glaucoma. In these 70 eyes, there were many locations where the sensitivity was significantly higher with SS than with FT (intrasubject difference), and SS had less intersubject variability than FT at most locations. The cumulative decibel scores at the region of glaucomatous defects were larger with SS (206.2+/-103.3 dB) than with FT (162.1+/-87.5 dB) (p=0.02), which indicated that the depth of defects measured by SS was shallower than that by FT. The sizes of defects were significantly larger with SS (11.2+/-5.6) than with FT (9.7+/-5.1) (p<0.05). CONCLUSIONS: Glaucomatous defects were measured as being significantly shallower and larger with SS than with FT. In addition, the prevalence of visual field defect was higher with SS according to some of the criteria for glaucomatous visual field defects. These results might be related to the fact that SS strategy has a lower variability and to the Bayesian statistical properties of the SITA algorithm.

Dominant optic atrophy in a Japanese family with OPA1 frameshift mutation (V942fsX966).

PURPOSE: The authors report the ophthalmic characteristics of a male proband in a Japanese family with autosomal dominant optic atrophy (DOA) harboring a frameshift mutation in the OPA1 gene. METHODS: Conventional ophthalmologic examinations including static automated perimetry were performed, as well as assessment of the three-generation family history. The peripapillary retinal nerve fiber layer (RNFL) was evaluated using scanning laser polarimetry. Mutation screening of the OPA1 gene was performed with polymerase chain reaction amplification and direct sequencing. RESULTS: A frameshift mutation (p.V942fsX966) was identified in the proband and his mother. In comparison with the adolescent onset of visual loss in the proband and his maternal grandfather, the mother presented with only subtle temporal disc pallor and has never been aware of any visual disturbances. Symmetric thinned peripapillary RNFL was detected in the proband, whose visual field abnormalities were limited to central scotomas and were without mean deviation worsening between 11 to 17 years of age in both eyes. The proband's logMAR visual acuity (0.52 to 0.7) has remained almost unchanged for more than 10 years since initial evaluation at age 10. CONCLUSIONS: The OPA1 mutation may be correlated with slow progression of DOA, and with phenotypic variations within the family. Further study is necessary to determine whether symmetric thinned peripapillary RNFL represents a feature of DOA.

Comparison of telomerase and CD44 expression as diagnostic tumor markers in lesions of the thyroid.

The analysis of the tissue expression patterns of both the telomerase enzyme and the adhesion molecule CD44 has highlighted these molecules as potential tumor markers. In this study, the expression of these markers was analyzed in frozen tissue samples of the same human thyroid lesions, and the data were compared to evaluate their application to tumor diagnosis. The study analyzed 12 malignant specimens, including 5 papillary, 3 follicular, 2 anaplastic, 1 medullary, and 1 low-grade Hurthle cell carcinoma and 17 specimens from benign lesions, including cases of adenoma, hyperplasia, and Graves' disease. Telomerase expression was analyzed by assay of enzyme activity using the telomeric repeat amplification protocol and by reverse transcription-PCR detection of human telomerase reverse transcriptase (hTERT) mRNA. Nine of 12 (75%) malignant samples and the two Graves' disease samples were evaluated as positive for telomerase activity by the telomeric repeat amplification protocol assay. The presence of hTERT mRNA was detected in 8 (67%) of 12 malignant tissues and in 5 (29%) of 17 benign thyroid tissue samples. The expression of CD44 transcripts containing variant exons 7, 8, and 11 was evaluated by reverse transcription-PCR/Southern blot analysis. Of the 12 malignant samples, 9 (75%) included transcripts containing exon 7, 10 (83%) included transcripts containing exon 11, and 11 (92%) included transcripts containing exon 8. However, these CD44 exons were also present in transcripts in a high proportion of benign samples. Five (28%), 10 (59%), and 6 (35%) benign samples contained CD44 transcripts, including variant exons 7, 8, and 11, respectively. The measurement of telomerase activity proved to be the most specific for the detection of thyroid carcinoma in frozen tissue samples as a single analyte, but diagnostic accuracy was increased by the combination of telomerase and CD44 analyses.

Telomerase activity in lesions of the thyroid: application to diagnosis of clinical samples including fine-needle aspirates.

The telomerase enzyme is capable of replacing telomeric DNA sequences that are lost at each cell division. It has been suggested that the function of this enzyme is necessary for cells to become immortal, and in concordance with this hypothesis, telomerase activity has been detected in malignant tumor cells, whereas the enzyme is inactive in normal somatic cells. The measurement of this activity in human tissue samples may have diagnostic value, and in this study, we examined whether such a measurement may be useful for the detection of malignant cells within the thyroid. Telomerase activity was assayed using the telomeric repeat amplification protocol and related to the histological diagnosis of thyroid biopsy tissue samples and of cells obtained from the thyroid by fine-needle aspiration (FNA). Extracts from 9 of 11 (82%) carcinoma biopsy tissue samples contained telomerase activity, whereas enzyme activity was detected in only 2 of 14 (14%) benign tissue sample extracts. These two positive cases were subsequently diagnosed as Graves' disease with severe lymphocytic infiltration. Five of six (83.3%) histologically confirmed carcinoma FNA samples were identified by using the telomeric repeat amplification protocol assay, and two samples considered to be suspicious by FNA cytology were also positive. Conversely, only 4 of 48 (8.3%) benign FNA samples had telomerase. These promising data indicate that this sensitive assay could become a useful adjunct to microscopic cytopathology in the detection of cancer cells in small tissue biopsies and in fine-needle aspirates of the thyroid.

Optical coherence tomography in enhanced S-cone syndrome: large macular retinoschisis with disorganized retinal lamination.

PURPOSE: The authors previously reported clinical and molecular features of a boy (at age 17) with enhanced S-cone syndrome (ESCS) who had bilateral cystoid maculopathies. The purpose of the current study was to describe the patient's optical coherence tomography (OCT) findings. METHODS: OCT was performed when the patient was 23 years old. RESULTS: OCT images revealed formations of single large hyporeflective cystic spaces in the cystoid maculopathies of both eyes. The cystic spaces were much larger than those of previously reported cases. In the temporal region of each cystic space, symmetric disorganized retinal lamination was observed with the retina lacking well-defined and hyporeflective bands of the inner nuclear layer and the outer nuclear layer, which are seen in normal retina. Splitting in the retinal thickness was at the level of the outer retinal layer rather than at the inner retinal layer in both eyes. CONCLUSIONS: Splitting is likely to occur close to the outer plexiform layer in which the cleavage plane of familial juvenile retinoschisis is identified. The unique OCT manifestation of symmetric large macular retinoschisis with disorganized retinal lamination may indicate a severe form of ESCS.

The expression of stat91, ras-independent signal transducer and activator, in human gastric carcinomas.

Upon binding of interferon (IFN) and epidermal growth factor (EGF) to their cell surface receptors, tyrosine phosphorylation of latent cytoplasmic Stat91 (Ras-independent signal transducer and activator of transcription, Stat1 alpha) protein is promptly induced and translocate from the cytoplasm to the nucleus to transduce the signal. The expression of mRNA for Stat91 was examined in 8 gastric carcinoma cell lines and 21 gastric carcinoma tissues as well as corresponding normal mucosa. Of the 8 gastric carcinoma cell lines, all expressed a 4.7 kb Stat91 mRNA and a 91 kD protein at various levels. In gastric carcinoma cell lines, the levels of Stat91 mRNA expression were compatible with those of Stat91 protein expression. In surgical cases, all the gastric carcinoma tissues and their adjacent non-neoplastic mucosa expressed Stat91 mRNA and protein. Interestingly, 14 (66%) out of 21 tumors expressed Stat91 mRNA at higher levels than their corresponding normal mucosas. Moreover, 6 (75%) of 8 tumor tissues expressed higher levels of Stat91 protein as compared with those of the corresponding normal gastric mucosa. No significant correlation was detected between the expression of Stat91 and clinicopathological feature of gastric carcinoma. These results suggest that the majority of gastric cancer in vivo harbour overexpression of Stat91 as a signal transducer in response to various cytokines or growth factors which may be implicated in the growth of gastric cancer.

Changes in descemet membrane and endothelium after corneal epithelial abrasion alone and with photorefractive keratectomy in rabbits.

OBJECTIVES: To investigate the effects of epithelial abrasion on the corneal endothelium and Descemet membrane in rabbits and to compare the changes with those after excimer laser photoablation of the cornea. METHODS: Central epithelial abrasions, 6 mm in diameter, were created by mechanical removal of the cells, and the specimens were examined from 24 hours up to 30 days by transmission electron microscopy. Corneas that were photoablated by an excimer laser and nontreated normal corneas were investigated as controls. RESULTS: Corneas denuded of epithelium showed massive enlargement of the mitochondria in the endothelium and exhibited a layer of electron-dense fibrillogranular material that had migrated forward through the Descemet membrane. These alterations were similar to the changes observed after photoablation of the cornea by an excimer laser. CONCLUSION: It was postulated that the extrusion of electron-dense material in the Descemet membrane observed after excimer laser ablation might have occurred primarily not as a result of shock waves, but from destruction of the epithelial integrity.

Inhibition of factor Xa suppresses the expression of tissue factor in human monocytes and lipopolysaccharide-induced endotoxemia in rats.

BACKGROUND: Activated factor X (FXa) is involved in hemostasis, thrombogenesis, inflammation, and cellular immune response. Tissue factor (TF) is an initiator of blood coagulation. We investigated whether FXa induces TF expression in human peripheral monocytes and whether treatment with FXa inhibitor reduces TF expression in an experimental model of rat endotoxemia. METHODS: Human peripheral mononuclear cells were used to determine TF expression induced by FXa. Experimental rat endotoxemia was induced by intravenous bolus injection of lipopolysaccharide (LPS). A specific FXa inhibitor, DX-9065a, was administered subcutaneously immediately after LPS injection. RESULTS: FXa induced TF expression in monocytes without intervention of thrombin and the expression was suppressed by FXa inhibitor. In the experimental model of rat endotoxemia, TF and TF mRNA expression levels in the liver were reduced by DX-9065a. Moreover, administration of DX-9065a suppressed the rise in plasma concentrations of thrombin-antithrombin III complex (TAT) and monocyte chemoattractant protein-1 (MCP-1). CONCLUSIONS: Our results indicated that FXa can induce TF expression in human peripheral monocytes and that inhibition of FXa reduces TF expression in the liver of rat endotoxemia. These results suggest that FXa is an important factor for TF expression in sepsis.

Expression of cyclin D1, CDK4 and p27KIP1 is associated with the p16MTS1 gene status in human esophageal carcinoma cell lines.

p16MTS1/INK4A negatively regulates cell cycle progression by inhibiting the cyclin D/CDK4 complex that phosphorylates pRb. Frequent homozygous deletions of the p16 gene were recently found in various tumor cell lines. We examined the relationship between the genetic status of p16 and the expression of the cell cycle regulating molecules in human esophageal carcinoma cell lines. Out of eight human esophageal carcinoma cell lines, seven (67.5%) and six (75%) cell lines showed homozygous deletions of the p16 and p15 genes, respectively. All the p16-negative cell lines expressed high levels of cyclin D1, CDK4 and p27KIP1 proteins. Interestingly, the expression level of cyclin D1 was closely correlated to the levels of not only CDK4 but also p27KIP1 protein in p16-negative cell lines. Furthermore, all the p16-negative cell lines expressed Rb protein of approx 110 kDa which corresponds to the phosphorylated form, whereas the cell line with intact p15 and p16 genes did not express pRb. These results suggest that the expression of cyclin D1, CDK4, Rb and p27 is associated with the p16 gene status in esophageal carcinoma cell lines. Alternatively, loss of the p16 gene and subsequent over-expression of cyclin D1 and CDK4 might be involved in autonomous growth of esophageal carcinoma cells.

Anti-tremor activity of talipexole produced by selective dopamine D2 receptor stimulation in cynomolgus monkeys with unilateral lesions in the ventromedial tegmentum.

The anti-tremor activity of talipexole (6-allyl-2-amino-5,6,7,8-tetrahydro-4H-thiazolo[4,5-d]azepine dihydrochloride, B-HT 920 CL2, Domin), a non-ergot dopamine D2 receptor agonist which possesses alpha 2-adrenoceptor agonistic and 5-HT3 receptor antagonistic properties, was examined in monkeys with a unilateral lesion in the ventromedial tegmentum. Talipexole dose dependently suppressed the tremor and had ED50 values of 34 micrograms/kg s.c. and 84 micrograms/kg p.o. The anti-tremor effect of talipexole occurred at much lower doses than that of an ergot dopamine receptor agonist, bromocriptine (2-bromo-alpha-ergocryptine mesilate, ED50; 2.5 mg/kg s.c.), and talipexole acted synergistically in combination with L-DOPA (3,4-dihydroxyphenylalanine). In ventromedial tegmentum-lesioned monkeys, anti-tremor doses of talipexole did not cause emetic behavior, but had sedative effects. Conversely, monkeys given bromocriptine exhibited oral movement, salivation and vomiting when anti-tremor effects were observed, but not marked sedative behavior at any of the doses investigated. During repeated administration of talipexole (a daily dose of 50 micrograms/kg s.c. for 21 days), the extent and duration of the anti-tremor effect did not change, but those of the sedative effect decreased gradually. The anti-tremor effect of talipexole was significantly suppressed by sulpiride, but not by SCH 23390 (7-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3-benzazepine-7-ol) or yohimbine, while the sedative effect was inhibited by sulpiride and yohimbine. The main metabolites of talipexole had no anti-tremor or sedative effects. These results indicate that talipexole exerts its anti-tremor activity via selective dopamine D2 receptor stimulation.

Diffuse ganglioneuromatosis with plexiform neurofibromas limited to the gastrointestinal tract involving a large segment of small intestine.

A rare case of diffuse ganglioneuromatosis with plexiform neurofibromas limited to the gastrointestinal tract in a 20-year-old man is reported. The patient had no cafe-au-lait spots or any apparent tumorous lesions affecting other organs. A fan-shaped resection of the mesentery, including 200cm of the ileum, together with ileocecal resection was done, and the diagnosis was histologically confirmed. The relationship between this benign tumor and other neurogenic disorders is discussed, along with a review of the literature. Long-term follow up of this patient is required because of the possible development of von Recklinghausen's disease or multiple endocrine neoplasia type IIb.

A model for oscillating chemical reactions.

A simple theoretical model for a class of oscillating chemical reactions is investigated, which is inspired by Degn's analysis of the mechanism of Belousov-Zhabotinsky reactions. Although the model involves only one auto-catalytic path, as distinct from models presented hitherto, a limit-cycle behaviour is found for the concentrations, corresponding to a definite region in the space of controllable parameters. Characteristics of the coherent oscillations are investigated together with the behaviour in the induction period. The critical slowing-down around the onset of instability is shown to behave very much like that found in the equilibrium phase transition. The existence of an "anti-Curie" point is found as well as the Curie point, corresponding to the fact that the domain for sustained oscillation is closed in certain directions. Possible extensions of the model and ways of improving the theory are discussed.

Expression of e-cadherin, alpha-catenins and Beta-catenins in human gastric carcinomas - correlation with histology and tumor progression.

The expression of cell-cell adhesion molecule, E-cadherin and its associated proteins, alpha- and beta-catenins in human gastric carcinomas was examined by Western blotting. All the seven gastric carcinoma cell lines expressed E-cadherin except KATOIII, which was derived from pleural effusion of a scirrhous type stomach cancer or Borrmann's type-4 carcinoma. The expression of alpha-catenin was not detected in HSC43 derived from scirrhous carcinoma, while HSC39 expressed abnormal beta-catenin caused by genetic alteration. In gastric carcinoma cases, the levels of E-cadherin and alpha-catenin were significantly lower in poorly differentiated adenocarcinomas and scirrhous carcinomas when compared to other types of gastric carcinomas. Deeply invasive carcinomas expressed E-cadherin and alpha-catenin at lower levels. However, the expression level of alpha-catenin was not necessarily consistent with that of E-cadherin. One of 10 gastric carcinomas examined showed complete deletion of alpha-catenin gene in Southern blotting. beta-catenin was expressed at lower level in poorly differentiated adenocarcinomas than in well-differentiated adenocarcinomas. These findings suggest that reduction in the expression of E-cadherin and its associated molecules, catenins, is involved in the development and infiltrative growth of scirrhous type gastric carcinomas.

Increased expression and phosphorylation of sh-ptp2 (syp) in human gastric carcinomas.

The expression of SH-PTP2 (SYP), a novel protein tyrosine phosphatase with src homology 2, was examined in human gastric carcinoma cell lines and gastric carcinoma tissues as well as corresponding non-neoplastic mucosas by Northern and Western blotting. The expression of SH-PTP2 mRNA was detected in all 8 gastric carcinoma cell lines at various levels. Most of the cell lines expressed SH-PTP2 protein of 70 kd which corresponds to tyrosine-phosphorylated form, whereas only one cell line expressed unphosphorylated form of SH-PTP2; Fourteen (66.6%) of 21 gastric carcinomas expressed SH-PTP2 mRNA at higher level than non-neoplastic mucosas. Furthermore, most of the gastric carcinomas displayed higher levels of tyrosine-phosphorylated form of SH-PTP2, while non-neoplastic mucosas tended to express unphosphorylated SH-PTP2 protein. These results suggest that the increased expression and tyrosine-phosphorylation of SH-PTP2 may participate in stomach carcinogenesis.

Expression of Trk tyrosine kinase receptor is a biologic marker for cell proliferation and perineural invasion of human pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is a widely known severe malignancy with a poor prognosis. Perineural invasion extending to the extra-pancreatic nerve plexus, a significant concern in the treatment is frequently present in this cancer. We analyzed immunohistochemical expression of neurotrophins (NGF, BDNF, NT-3) and the cognate receptors, Trk tyrosine kinase receptor family (TrkA, B, C) and p75NGFR in 28 surgically resected PDAC specimens. A comparative study between several clinicopathologic factors and Trk receptors revealed a significant correlation between increased expression of TrkA and cancer proliferation, as well as TrkC and cancer invasion, including venous and perineural invasion. The present findings revealed a novel mechanism in PDAC progression that is mediated via a NTs-Trk interaction.

Molecular cloning, expression and tissue distribution of hamster diacetyl reductase. Identity with L-xylulose reductase.

Using rapid amplification of cDNA ends PCR, a cDNA species for diacetyl reductase (EC 1.1.1.5) was isolated from hamster liver. The encoded protein consisted of 244 amino acids, and showed high sequence identity to mouse lung carbonyl reductase and hamster sperm P26h protein, which belong to the short-chain dehydrogenase/reductase family. The enzyme efficiently reduced L-xylulose as well as diacetyl, and slowly oxidized xylitol. The K(m) values for L-xylulose and xylitol were similar to those reported for L-xylulose reductase (EC 1.1.1.10) of guinea pig liver. The identity of diacetyl reductase with L-xylulose reductase was demonstrated by co-purification of the two enzyme activities from hamster liver and their proportional distribution in other tissues.