Detection and identification of oxidants formed during •NO/O2•⁻ reaction: a multi-well plate CW-EPR spectroscopy combined with HPLC analyses.

New techniques and probes are routinely emerging for detecting short-lived free radicals such as superoxide radical anion (O2(•-)), nitric oxide ((•)NO), and transient oxidants derived from peroxynitrite (ONOO(-)/ONOOH). Recently, we reported the profiles of oxidation products (2-hydroxyethidium, ethidium, and various dimeric products) of the fluorogenic probe hydroethidine (HE) in the (•)NO/O2(•-) system (Zielonka et al. 2012). In this study, we used HPLC analyses of HE oxidation products in combination with continuous wave electron paramagnetic resonance (CW-EPR) spin trapping with 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO) to define the identity of the oxidizing species formed in the (•)NO/O2(•-) system. EPR spin-trapping technique is still considered as the gold standard for characterization of free radicals and their intermediates. We monitored formation of BMPO-superoxide (BMPO-(•)OOH) and BMPO-hydroxyl (BMPO-(•)OH) radical adducts. Simultaneous analyses of results from EPR spin-trapping and HPLC measurements are helpful in the interpretation of the mechanism of formation of products of HE oxidation.

Evaluation of malignancy and the prognosis of esophageal cancer based on an immunohistochemical study (p53, E-cadherin, epidermal growth factor receptor).

The subjects in this study consisted of 40 preoperative untreated esophageal squamous cell carcinoma patients. While p53 did not significantly correlate with the clinicopathological factors, E-cadherin significantly correlated with lymphatic invasion, vascular invasion, the depth of invasion, the degree of lymph node metastasis, the histological stage, and the number of lymph node metastases. Epidermal growth factor receptor (EGFR) significantly correlated with age, the depth of invasion, and the number of lymph node metastases. The 5-year cumulative survival rate was 45.7% in the p53-positive cases and 61.9% in the p53-negative cases, with no significant difference, and 87.8% in the E-cadherin-positive cases and 19.1% in the -negative cases, and the difference was significant. The prognosis was significantly poor in EGFR-positive subjects: the 5-year survival rate was 38.6% in EGFR-positive cases and 68% in -negative cases. The 5-year survival rate in E-cadherin-negative, EGFR-positive cases was 0%, while it was 91.7% in the reverse pattern, and this difference was significant. These findings suggest that both E-cadherin and EGFR are important prognostic factors, and a more precise prognosis can thus be obtained by combining them. Such a combined technique may be very useful as an indicator for grading the biological malignancy of esophageal cancer.

[Slow releasing anticancer drug containing CDDP for intraoperative use in residual cancer cells].

We have already reported the slow releasing property and anticancer effect of Plachitin, which is reconstituted by combination of CDDP and chitin. This study deals in more detail with the slow releasing property, and the renal complications and the effectiveness for solid tumor were examined. After implantation of Plachitin subcutaneously in the abdominal wall, the platinum concentration in the different organs was measured. In the abdominal muscle around the implanted Plachitin, a high concentration of platinum was maintained until 8 weeks and the peak was 4 weeks after implantation. At the same time, the serum concentration of platinum remained low. In kidney, the platinum concentration resembled the levels in the abdominal muscle, but no renal dysfunction was found serologically or histologically. When Plachitin was implanted around the solid tumor, the survival rates were improved and the gain in tumor weight was suppressed as compared with the controls. From these findings, Plachitin seemed to be effective as a slow releasing anticancer drug for topical application.

[Development of slow releasing anticancer drug based with absorbable biomaterial chitin].

To have a comparatively more slowly releasing anticancer drug with effectiveness, Plachitin was prepared by chemical combination of CDDP and chitin (poly-N-acetyl-D-glucosamine). Chitin is absorbed by the living body over several months. To investigate the slow releasing property, it was implanted in thigh muscle of mice and rabbit. Pt level in different organs and in urine was measured at regular intervals. Pt level in implanted muscles was higher in comparison to low serum level in mice. It was released slowly over 1 to 2 months in mice, whereas in rabbit it took about three weeks. Pt releasing period of the Plachitin was different according to the adopted method of implantation. Anticancer effect of Plachitin was investigated by injecting 180 sarcoma cells in mouse peritoneal cavity and subsequent implantation of Plachitin. In control groups chitin was used instead of Plachitin. The survival rate of mice in the Plachitin group after 14 days was higher than in the chitin group, and the anticancer effect of the Plachitin was confirmed.

[Drug concentration in cancerous large bowel tissue and thymidylate synthase inhibition rate after administration of tegafur and UFT].

Tegafur and UFT are most widely used for the treatment of cancers of the gastrointestinal tract. FdUMP, a metabolite of 5-fluorouracil(5-FU), is known to have an inhibitory activity of thymidylate synthase (TS). We administered 600 mg/day of tegafur or UFT to 37 patients with cancer of the large bowel beginning 1 week prior to surgery and then measured the concentrations of tegafur and 5-FU, and TS inhibition rate in the excised tissue. Our results showed that the concentrations of 5-FU in tumor were 0.077 +/- 0.026 microgram/g in tegafur group and 0.208 +/- 0.143 microgram/g in UFT group, with the UFT showing significantly higher levels (p less than 0.05). On the other hand, TS inhibition rates in tumor tissue were 45.5 +/- 13.3% in tegafur group and 56.1 +/- 13.0% in UFT group, with a significantly higher level existing in the latter group (p less than 0.05). Furthermore, the same high 5-FU concentration and TS inhibition rates were observed in the lymph nodes affected by metastasis. No difference between the tegafur and UFT groups was noted in normal tissue or normal lymph nodes, and compared to tegafur, UFT showed an effective action on cancerous large bowel tissue.

Cybex II Hip and Trunk Stabilization during Quadriceps/Hamstring Evaluation.

The purpose of this paper is to suggest a restraint system that can be used during the use of a Cybex II in testing the quadriceps and hamstrings. Information suggests that torque values can be affected without the use of a restraint system. J Orthop Sports Phys Ther 1981;2(4):191-193.