Detection of gastric cancer-associated microRNAs on microRNA microarray comparing pre- and post-operative plasma.

BACKGROUND: Recently, it was reported that plasma microRNAs (miRNAs) are low-invasive useful biomarkers for cancer. We attempted to isolate gastric cancer (GC)-associated miRNAs comparing pre- and post-operative paired plasma, thereby excluding the possible effects of individual variability. METHODS: This study was divided into four steps: (1) microarray analysis comparing pre- and post-operative plasma; (2) validation of candidate miRNAs by quantitative RT-PCR; (3) validation study of selected miRNAs using paired plasma; and (4) comparison of the levels of selected miRNAs in plasma between healthy controls and patients. RESULTS: From the results of microarray analysis, nine candidate miRNAs the levels of which were markedly decreased in post-operative plasma were selected for further studies. After confirmation of their post-operative marked reduction, two candidate miRNAs, miR-451 and miR-486, were selected as plasma biomarkers, considering the abundance in plasma, and marked decrease in post-operative samples. In validation, the two miRNAs were found to decrease in post-operative plasma in 90 and 93% of patients (both P<0.01). In comparison with healthy controls, the levels of both miRNAs were found to be significantly higher in patients, and the area under the curve values were high at 0.96 and 0.92. CONCLUSION: Plasma miR-451 and miR-486 could be useful blood-based biomarkers for screening GC.

Differential expression and pathological significance of autocrine motility factor/glucose-6-phosphate isomerase expression in human lung carcinomas.

To clarify the involvement of autocrine motility factor (AMF) in the phenotype and biological profiles of human lung carcinomas, we analysed protein and mRNA expression in a total of 180 cases. Immunohistochemistry revealed positive staining in 67.2%, with the highest frequency in squamous cell carcinoma (SCC; 90.8%) and the lowest in small cell carcinoma (SmCC; 27.8%). In SCC, the staining frequency and intensity correlated with the degree of morphological differentiation. Generally, the expression levels in immunoblotting analysis corresponded well with immunohistochemical positivity. However, there was less agreement between protein and mRNA levels: in SmCC and large cell carcinomas (LCCs), mRNA showed higher, but protein showed lower expression. Among non-small cell lung carcinomas (NSCLCs), AMF protein levels correlated inversely with tumour size, but tumours exhibiting lymph node metastasis showed higher mRNA expression. In cultured lung carcinoma cells which comprised all histological subtypes, AMF was detected in the lysates of all ten cell lines. Secreted AMF protein was detected in the conditioned media from six cell lines, most of which were SmCC or LCC. Thus, a particular subset of lung carcinomas secrete AMF, which may promote cell motility via autocrine stimulation through its cognate receptor and cause the biological aggressiveness seen in SmCC and LCC. Moreover, treatment by proteasome inhibitors resulted in increased cellular AMF in five cell lines, suggesting that intracellular AMF levels are regulated by both secretion and proteasome-dependent degradation. In conclusion, AMF was detected in a major proportion of lung carcinomas, and may play a part not only in proliferation and/or progression of the tumours, but also, possibly, in the differentiation of SCC. Furthermore, higher mRNA expression may be related to the high metastatic potential of NSCLC and increased protein secretion, leading to a more aggressive phenotype, such as the invasiveness of SmCC and LCC.

Surgical treatment of abdominal aortic aneurysm associated with horseshoe kidney and coagulopathy case report.

The coexistence of horseshoe kidney and aortic aneurysm poses a technical challenge to the vascular surgeon during aneurysm repair. Whether to divide the renal isthmus and how to approach the aneurysm are still matters of controversy, and coagulopathy sometimes occurs in patients with nontreated abdominal aortic aneurysm (AAA). We describe the successful surgical repair of an AAA with horseshoe kidney via the transperitoneal approach and division of the renal isthmus by harmonic scalpel. Exclusion of a thrombosed aneurysm can ameliorate coagulopathy due to AAA.

An experimental model of osteoarthrosis of the temporomandibular joint in monkeys.

The purpose of this study was to develop a model of osteoarthrosis of the temporomandibular joint in monkeys, which is remarkably similar in structure and function to that of humans. Nine juvenile monkeys, two as controls and seven as an experimental group, were used in this study. In the experimental group, the articular eminence on both sides was surgically made steeper. Two animals were killed at 1 week, four at 6 months, and one at 1 year postoperatively and the temporomandibular joints were examined macroscopically and microscopically. Typical changes of osteoarthrosis were observed in the 6-month and 1-year specimens. These comprised clustering of chondrocytes which resulted in vertical and horizontal splitting in the articular cartilage, and fibrillation of the articular surface resulting in fibrous union in the joint cavity. These degenerative changes advanced progressively over time. Slight anterior displacement and degenerative changes in the articular disc were also seen.

[Recognition on the gender of point-light walkers moving in different directions].

When we see a series of moving lights attached to a walker's major joints, we can recognize it as a human body in motion (Bruce & Green, 1990; Johansson, 1973). Moreover, observers can recognize the walker's gender from such a display (Barclay, Cutting, & Kozlowski, 1978; Cutting & Kozlowski, 1977; Cutting, Proffitt, & Kozlowski, 1978). For gender recognition, Cutting and his colleagues suggested that male and female walkers respectively have unique walking styles caused by differences in the position of their center of moment (Cm). However, previous studies of gender recognition used only side views of these point-light walkers. The present study investigated gender recognition by presenting subjects with three kinds of views of the point-light walkers: profile views of rightward movements, approaching movements, and 3/4 profile diagonal movements. Subjects' task was to judge if the walker was male or female. The results showed almost the same judgement accuracy for all directions in male walkers, and the highest accuracy for the approaching movement in female walkers. The subjects tended to judge the presented walkers to be male more frequently than female.

Development of postoperative delirium in relation to a room change in the general surgical unit.

To analyze the relationship between the development of postoperative delirium and a change of the patient's room, 1,006 cases of patients who had undergone surgery with general anesthesia were reviewed. Postoperative delirium developed in 84 (8.3%) cases. On the basis of symptomatic features, postoperative delirium was divided into four types: (1) excitement type, (2) excitement-hallucination type, (3) hallucination type, and (4) disorientation type. Of the 31 excitement-type cases, 21 developed within the 2nd postoperative day (POD) while 27 of 29 hallucination types developed after POD 2. Of 29 hallucination types, 22 developed after a room change while 20 of these 22 cases were transferred to a single room before POD 2. A quiet, dark, and isolated environment in a single room is suggest to contribute to the development of hallucinations. The development of postoperative delirium with hallucinations alone should thus be taken into consideration whenever a room change is decided.

Production of functional S-II in Bombyx mori cells.

A cDNA for murine transcription factor S-II (Hirashima et al., J. Biol. Chem (1988) 263, 3858-3863) was inserted into a silkworm baculovirus vector and expressed in Bm-N cells, a cell line of Bombyx mori maintained in the laboratory. Recombinant S-II was purified from virus-infected cell extracts to near homogeneity by ammonium sulfate fractionation, and chromatographies on DEAE-cellulose and phosphocellulose. About 60 micrograms of recombinant S-II was obtained from 1 g of virus-infected cells. The molecular mass and specific activity of recombinant S-II were exactly the same as those of authentic S-II purified from Ehrlich ascites tumor cells.

Augmentation and stable expression of a novel transcription factor SII in CD4-positive cells on infection with human immunodeficiency virus type-1 (HIV-1).

We have examined the expression of a novel transcription factor SII in HIV-infected cells. Concomitant with the increasing expression of HIV-1 mRNA, the viability of HIV infected cells dropped and the expression of beta-actin gene also diminished. Concurrently, the expression of SII mRNA was stabilized or augmented. The data presented in this communication suggests that in HIV-1 infected cells, selective transcriptional controls were taking place and that a novel transcriptional factor, SII, plays an essential role in HIV-1 expression leading to cytocidal effects.

Molecular cloning and characterization of cDNA for eukaryotic transcription factor S-II.

Complementary DNA (cDNA) clones encoding a transcription factor S-II were isolated and characterized. The primary structure of S-II was determined by nucleotide sequence analysis of these clones. The predicted primary structure was consistent with the model that we proposed previously from the results of biochemical analyses of S-II. Using these clones as probes, we analyzed the mRNA for S-II. RNA blot analysis demonstrated the presence of four species of mRNA that hybridized with S-II cDNA in Ehrlich ascites tumor cells. This is the first evidence of polymorphism of mRNA encoding a transcription factor of RNA polymerase II. The results of analysis of the genomic structure suggested that the polymorphism of mRNA may be due to alternative splicing, or differences in initiation or termination of transcription.

A pedigree of Leber's congenital amaurosis.

A pedigree of Leber's congenital amaurosis compatible with autosomal recessive trait is reported. Two male infants from consanguineous parents had remarkable visual loss within the first year of life, with sluggish pupillary responses, poor fixations, minimal eyeground changes and absent electroretinograms on presentations at the ages of four or 14 months. Follow-up studies revealed definite progressions of eyeground abnormalities consisting of attenuated retinal arterioles, pepper- and salt-like appearance with numerous yellowish-white punctate lesions in the midperiphery, and pale optic nerves. Fluorescein angiographic study performed on one case showed multiple hyperfluorescent spots over the posterior and midperipheral eyegrounds suggesting alterations of the retinal pigment epithelium. These functional and morphological abnormalities of the retina were similar in the two siblings. Cycloplegic refractions revealed slight myopic or mixed astigmatism, but no marked hyperopia. The patients had normal physical and mental developments with no obvious systemic complications.

Orbital rhabdomyosarcoma: case report with immunohistochemical detection of muscle cell type intermediate filament proteins.

A 4 1/2-year-old girl had a rapidly growing tumor in the orbit. An extirpated mass consisted of small round or spindle neoplastic cells containing hyperchromatic mitotic nuclei. The cytoplasm had many lipids and glycogens with few organelles. A light and electron microscopic examination revealed an exceptional tumor cell with distinct cross striations compatible with typical rhabdomyoblast; occasional cells exhibited a few bundles of thin filaments scattered in the cytoplasm. Immunohistochemical studies for muscle cell-related proteins by avidin-biotin-peroxidase complex methods revealed positive reactions of the tumor cells to the antiserum against myoglobin, myosin, desmin and vimentin, providing evidence for muscle cell derivation of the neoplastic cells.

Phosphorylation of S-II is not affected by inhibitors of RNA synthesis.

S-II is an essential factor for RNA polymerase II-mediated transcription. A phosphorylated form of S-II, termed S-II has been shown to be present in cells at half the concentration of S-II. In studies on the role of phosphorylation and dephosphorylation of S-II in transcription, the possibility that phosphorylation of S-II is coupled with transcription in vivo was investigated. The phosphorylation of S-II was measured in mouse L cells cultured with two typical inhibitors of RNA synthesis. Neither of these inhibitors, 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) and actinomycin D, affected the phosphorylation of S-II under conditions where 75 to 98% of RNA synthesis was inhibited at the initiation and elongation step, respectively. These results indicate that the phosphorylation of S-II and transcription are independent processes.

Structural relationships of the three stimulatory factors of RNA polymerase II from Ehrlich ascites tumor cells.

The structural relationships of S-II, S-II', and S-I(b) stimulatory proteins of RNA polymerase II purified from Ehrlich ascites tumor cells were investigated. From analysis of the amino acid compositions and tryptic peptide maps of these proteins labeled with radioiodinated Bolton-Hunter reagent, it was concluded that S-I(b) is a part of S-II located at either the amino- or carboxyl-terminal and that only this region mainly contains radioiodinatable amino acid residues when labeled using 125I. On chymotryptic digestion, S-II was cleaved to 21- and 18-kDa fragments in the presence of DNA. The 21-kDa fragment was found to be sufficient for stimulation of RNA polymerase II. It was suggested that S-II' is formed by phosphorylation of S-II in the domain containing the 18-kDa fragment.