Background— Earlier studies have shown that adrenomedullin (AM), a potent vasodilator peptide, has a variety of cardiovascular effects. However, whether AM has angiogenic potential remains unknown. This study investigated whether AM gene transfer induces therapeutic angiogenesis in chronic hind limb ischemia. Methods and Results— Ischemia was induced in the hind limb of 21 Japanese White rabbits. Positively charged biodegradable gelatin was used to produce ionically linked DNA-gelatin complexes that could delay DNA degradation. Human AM DNA (naked AM group), AM DNA-gelatin complex (AM-gelatin group), or gelatin alone (control group) was injected into the ischemic thigh muscles. Four weeks after gene transfer, significant improvements in collateral formation and hind limb perfusion were observed in the naked AM group and AM-gelatin group compared with the control group (calf blood pressure ratio: 0.60±0.02, 0.72±0.03, 0.42±0.06, respectively). Interestingly, hind limb perfusion and capillary density of ischemic muscles were highest in the AM-gelatin group, which revealed the highest content of AM in the muscles among the three groups. As a result, necrosis of lower hind limb and thigh muscles was minimal in the AM-gelatin group. Conclusions— AM gene transfer induced therapeutic angiogenesis in a rabbit model of chronic hind limb ischemia. Furthermore, the use of biodegradable gelatin as a nonviral vector augmented AM expression and thereby enhanced the therapeutic effects of AM gene transfer. Thus, gelatin-mediated AM gene transfer may be a new therapeutic strategy for the treatment of peripheral vascular diseases.

The aim of the present study was to investigate the safety and immunological responses of personalized peptide vaccination in combination with oral administration of a 5-fluorouracil derivative (TS-1) in advanced gastric or colorectal carcinoma patients. Eleven patients (four with gastric cancer and seven with colorectal cancer) who failed to improve by prior TS-1-based chemotherapies were enrolled in this study. Peptides to be administered to patients were determined based on the presence of peptide-specific cytotoxic T lymphocyte ( (1) Cancer vaccines have emerged as a promising therapeutic approach, (2) but their clinical responses have been limited.(3) In order to develop a new treatment modality, we recently devised a new regime of peptide-based vaccination that consists of measuring pre-existing CTL precursors and IgG reactive to many kinds of vaccine candidates, followed by administration. (4)(5)(6)(7)(8) In addition, we recently reported the benefits of the combination of this type of peptide vaccination with a low dose of estramustine phosphate in hormone refractory prostate cancer patients.(9) This chemoimmunotherapy may break through the impasse in the clinical efficacy of cancer vaccines.TS-1 is an oral fluoropyrimidine derivative consisting of Tegafur (FT) and two modulators, 5-chloro-2,4-dihydroxypyrimidine (CDHP) and potassium oxonate (Oxo). FT is a prodrug of 5-fluorouracil (5-FU) and CDHP is a reversible competitive inhibitor of an enzyme involved in the degradation of 5-FU. Therefore, the FT-derivative 5-FU remains in tumor tissue a long time, resulting in an enhanced antitumor effect.(10,11) Indeed, TS-1 has been reported to be effective at prolonging the survival of patients with advanced gastric cancer.(12) Furthermore, TS-1 can be administered as an oral formulation that permits treatment on an outpatient basis, leading to improvement in the quality of life (QOL) of patients. In the present study, we investigated the safety and immunological responses of personalized peptide vaccination in combination with the oral administration of TS-1 in advanced gastric or colorectal carcinoma patients. Materials and MethodsPatients and eligibility criteria. The study protocol was approved by the Institutional Ethical Review Boards of Hokkaido University, Okayama University and Kurume University. Complete written informed consent was obtained from all patients at the time of enrolment. According to the protocol, patients were confirmed to be HLA-A24 or HLA-A2 positive and had a histologically confirmed lesion of gastric or colorectal carcinoma. In HLA-A2 + patients, their HLA-A2 genotypes were determined using sequence-specific oligonucleotide DNA typing after polymerase chain reaction. All patients had failed to improve by prior chemotherapy with TS-1. The other eligibility criteria included an age of 85 years or less, serum creatinine of less than 1.4 mg/dL, bilirubin of less than 1.5 mg / dL, a platelet count of 100 000 µL or more, hemoglobin of 8.0 g / dL or more, and total white blood cell count (W...

Background-Synchrotron radiation has been used to analyze crossbridge dynamics in isolated papillary muscle and excised perfused hearts with the use of x-ray diffraction techniques. We showed that these techniques can detect regional changes in rat left ventricle contractility and myosin lattice spacing in in situ ejecting hearts in real time. Furthermore, we examined the sensitivity of these indexes to regional ischemia. Methods and Results-The left ventricular free wall of spontaneously beating rat hearts (heart rate, 290 to 404 bpm) was directly exposed to brief high-flux, low-emittance x-ray beams provided at SPring-8. Myosin mass transfer to actin filaments was determined as the decrease in reflection intensity ratio (intensity of 1,0 plane over the 1,1 plane) between end-diastole and end-systole. The distance between 1,0 reflections was converted to a lattice spacing between myosin filaments. We found that mass transfer (mean, 1.71Ϯ0.09 SEM, nϭ13 hearts) preceded significant increases in lattice spacing (2 to 5 nm) during systole in nonischemic pericardium. Left coronary occlusion eliminated increases in lattice spacing and severely reduced mass transfer (PϽ0.01) in the ischemic region. Conclusions-Our results suggest that x-ray diffraction techniques permit real-time in situ analysis of regional crossbridge dynamics at molecular and fiber levels that might also facilitate investigations of ventricular output regulation by the Frank-Starling mechanism. Key Words: ischemia Ⅲ myocardial contraction Ⅲ myosin Ⅲ radiography D espite the history of studies on crossbridge dynamics, lower photon counts and poorer quality of diffraction patterns obtained from cardiac muscle than skeletal and insect flight muscles 1-3 have limited progress with cardiac muscle until recently. 4,5 Some of us used third-generation synchrotron radiation (SPring-8, Japan Synchrotron Radiation Research Institute) to determine x-ray diffraction patterns in excised, perfused rat hearts while moving systematically across the left ventricular (LV) equator from the epicardium through to the ventricular cavity. 6 X-ray diffraction patterns of cardiac muscle produce 2 equatorial-position reflections from the lattice-like arrangement of its protein elements. 2 Mass transfer of myosin heads to actin during contraction is inferred from a decrease in the integrated 1,0 reflection intensity (I 1,0 , lattice plane containing only thick myosin filaments) and an increase in 1,1 reflection intensity (I 1,1 , plane with thick myosin and thin actin filaments). 7 The myocardial intensity ratio (defined as I 1,0 /I 1,1 ) is minimal in the rigor state and maximal in a quiescent state. 1,2,6,8 Furthermore, the distance between 1,0 reflection peaks (d 1,0 spacing) represents the myosin lattice spacing, which is inversely related to sarcomere length in isolated fibers 5 as static myocytes maintain a constant cell volume. Whether decreases in myofilament spacing contribute to increasing Ca 2ϩ sensitivity and increased probability of crossbridge formation at longe...