Involvement of epithelial-mesenchymal transition in adenoid cystic carcinoma metastasis

The high frequencies of recurrence and distant metastasis of adenoid cystic carcinoma (AdCC) are significant obstacles for the long-term cure of patients with AdCC and emphasize the need for better understanding of the biological factors associated with these outcomes. To identify proteins that mediate AdCC metastasis, we established three AdCC cell lines expressing green fluorescent protein (GFP) from the ACCS cell line using orthotopic transplantation and in?vivo selection in nude mice: Parental ACCS-GFP, highly tumorigenic ACCS-T GFP and metastatic ACCS-M GFP. ACCS-GFP and ACCS-M GFP were subjected to DNA microarray analysis and the results were used for data mining studies. DNA microarray analysis revealed significantly altered biological processes in the ACC-M GFP cells, including events related to cell adhesion (three categories) and signaling (three categories). In particular, a significant down-regulation of cell adhesion molecules, such as cadherins and integrin subunits was observed. The loss of E-cadherin and integrins and the gain of vimentin in ACCS-M GFP cells were confirmed by immunoblotting. These results suggest that epithelial-mesenchymal transition (EMT) is a putative event in AdCC metastasis that induces tumor cell dissemination from the primary tumor site. In summary, in this study we established a useful nude mouse metastasis model which will enable further AdCC metastasis research and clinical treatment trials and we also provide evidence that EMT is significantly involved in the AdCC metastatic process.     

Characterization of dilution conditions for diesel nanoparticle inhalation studies

Diesel exhaust nanoparticles easily coagulate during transportation from the engine to the inhalation chamber, depending on concentrations and residence times. Although dilution is effective in suppressing coagulation growth of nanoparticles, volatile organic carbon (OC) evaporates as a result of dilution. Thus, the design of an inhalation facility to investigate the health effects of nanoparticle-rich exhaust is important. In this study, we determined the optimum dilution conditions in consideration of coagulation growth and evaporation of OC for inhalation studies of nanoparticle-rich diesel exhaust. We found that a short residence time prevented coagulation growth in the primary dilution tunnel after the primary dilution or before the diluted exhaust reached the inhalation chamber after the secondary dilution. However, due to the longer residence time in the inhalation chamber, the coagulation growth occurred in the inhalation chamber depending on secondary dilution ratio which controlled exposure dose (particle concentration in the inhalation chamber). We determined that the secondary dilution ratio for the high-concentration chamber should be around 4.5 times to prevent coagulation growth and to obtain the desired exposure dose. We also found that the loss of OC was relatively independent of the secondary dilution ratio when the secondary dilution ratio was more than 10 times because it seemed to reach a gas-particle equilibrium in the inhalation chamber. We therefore set the secondary dilution ratios for the middle- and low-concentration chambers to 13.5 and 40.5 times, respectively.     

Determination of procalcitonin concentration using the SphereLight 180 clinical auto-analyzer

BACKGROUND: Procalcitonin (PCT) is a biomarker for the diagnosis of sepsis and bacterial infection diseases. METHODS: A new fully automated SphereLight PCT (SL-PCT) assay system for PCT concentration in human serum or plasma by using SphereLight 180 (SL180, Olympus Corp.) analyzer was developed. The SL-PCT assay is based on chemiluminescent enzyme immunoassay. RESULTS: A linear dose response relationship was observed up to 200 ng/ml PCT concentration. The detection limit of PCT concentration was 0.06 ng/ml. Endogenous substances, anticoagulants, sodium fluoride and drugs did not interfere with assay results. There was a good correlation between the present method and the manual method in serum and plasma samples. CONCLUSIONS: These results indicate that the SL-PCT assay showed good performance in terms of the linearity, detection limit and precision. Use of this PCT measurement may improve the detection of sepsis and infectious disease.     

Quercetin protects embryonic chicken spermatogonial cells from oxidative damage intoxicated with 3-methyl-4-nitrophenol in primary culture

Diesel exhaust particles (DEP) are considered to be one of the most important air pollutants. In this study, the protective effect of quercetin, an antioxidant flavonoid, on oxidative damage of testicular cells was studied by analysis of the intracellular antioxidant system of embryonic chickens after treatment with 3-methyl-4-nitrophenol (PNMC) derived from DEP. Testicular cells from 18-day-old embryos were cultured in serum-free McCoys’5A medium and challenged with PNMC (10⁻⁷ to 10⁻⁵ M) alone or in combinations with quercetin (1.0 μg/ml) for 48 h. Results showed that exposure to PNMC (10⁻⁵ M) induced condensed nuclei and vacuolated cytoplasm, a decrease in testicular cell viability and spermatogonial cell number. Exposure to PNMC induced lipid peroxidation by an elevation of thiobarbituric acid reactive substances as well as decreasing glutathione peroxidation activity and superoxide dismutase activity. However, simultaneous supplementation with quercetin restored these parameters to the similar levels as the control. PNMC is therefore concluded to have induced the oxidative stress of the spermatogonial cells, which can be attenuated by combined quercetin treatment. Our results support the therapeutic use of quercetin in the prevention or treatment of the reproductive toxicity by environmental toxicant PNMC.     

Endoscopic examination for duodenal ulcer bleeding during transcatheter arterial embolization: analysis of two cases

Endoscopy is usually effective in treating duodenal ulcer bleeding, but depending on the lesion site and overall patient condition, hemostasis may be difficult to achieve with endoscopy alone. We described two patients with duodenal ulcer bleeding in whom endoscopic hemostasis was difficult. Immediately after transcatheter arterial embolization, endoscopic examination was used to confirm hemostasis and completing of the angiographic procedures.     

Effect of Dietary Fatty Acid Composition on Th1/Th2 Polarization in Lymphocytes

Background: It has become increasingly clear that polyunsaturated fatty acids (PUFAs) have immunomodulatory effects. However, the intake of these fatty acids used in animal studies often greatly exceeds dietary human intake. Whether differences in the composition of fatty acids that are consumed in amounts consistent with normal dietary intake can influence immune function remains uncertain. Methods: We manufactured 3 types of liquid diet, related to modified fatty acid composition (ω‐6/ω‐3 = 0.25, 2.27 and 42.9), but excluding eicosapentaenoic acid and docosahexaenoic acid, based upon a liquid diet used clinically in humans. We assessed CD3‐stimulated cytokine production of splenocytes in female BALB/c mice (n = 4 per group) fed 1 of 3 liquid diets for 4 weeks. We also measured the cytokine production of peripheral blood mononuclear cells stimulated with phorbol myristate acetate and ionomycin in humans at the end of a 4‐week period of consumption of 2 different liquid diets (ω‐6/ω‐3 = 3 and 44). Results: We found that the ratio of interfero ω‐γ (IFN‐γ) / interleukin‐4 (IL‐4) was significantly higher in mice fed theω ‐3 rich diet than in others. In humans, IFN‐γ / IL‐4 was significantly higher after the ω‐3 versus the ω‐6 enhanced diet. Conclusions: Differences in the composition of ω‐3 andω ‐6 PUFAs induces a shift in the Th1/Th2 balance in both mouse and human lymphocytes, even when ingested in normal dietary amounts. An ω‐3 rich diet containing α‐linolenic acid modulates immune function.     

Changes in molecular markers of hemostatic and fibrinolytic activation under various sampling conditions using vacuum tube samples from healthy volunteers

Molecular makers such as thrombin-antithrombin complex (TAT), prothrombin fragment 1 + 2 (F1 + 2), soluble fibrin (SF), and D-dimer, are useful markers in the diagnosis and assessment of various thrombotic conditions. These markers are measured in plasma after blood sampling. Difficult blood sampling is known to falsely elevate plasma TAT levels. However, it is not known exactly why this occurs. In the present study, we examined how levels of molecular markers of haemostatic and fibrinolytic activation change under various sampling conditions using vacuum tube samples from healthy volunteers.When blood was sampled continuously by taking 10 consecutive vacuum tube samples following application of a tourniquet, blood sampling resulted in an accurate assessment of these molecular makers.When blood was sampled continuously by taking vacuum tube samples every one minute over a total of 9 minutes to investigate possible changes in the levels of the molecular markers over time, plasma levels of TAT, SF, and F1 + 2 gradually increased with time. Plasma levels of TAT, F1 + 2, and SF increased beyond the normal range over the course of nine minutes. When blood was sampled using three alternative methods, which varied in terms of the duration of needle puncture (sampling B), duration of tourniquet use (sampling C), or both (sampling A), plasma TAT and SF levels were significantly increased with all three methods, compared to control samples. Plasma F1 + 2 levels were significantly increased with sampling methods A and B, compared to control samples, but not with sampling method C. On the other hand, plasma D-dimer levels were not significantly altered by any of the sampling methods.In conclusion, the results suggest that molecular markers of haemostatic and fibrinolytic activation, except for D-dimer, may be affected by sampling method, particularly the duration of needle puncturing. Therefore, care needs to be taken when using TAT, F1 + 2, and SF levels to diagnose and estimate activation of the coagulation system.