Targeted Anticancer Immunotoxins and Cytotoxic Agents with Direct Killing Moieties

Despite the progress of the bioinformatics approach to characterize cell-surface antigens and receptors on tumor cells, it remains difficult to generate novel cancer vaccines or neutralizing monoclonal antibody therapeutics. Among targeted cancer therapeutics, biologicals with targetable antibodies or ligands conjugated or fused to toxins or chemicals for direct cell-killing ability have been developed over the last 2 decades. These conjugated or fused chimeric proteins are termed immunotoxins or cytotoxic agents. Two agents, DAB389IL-2 (ONTAKTM) targeting the interleukin-2 receptor and CD33-calicheamicin (Mylotarg®), have been approved by the FDA for cutaneous T-cell lymphoma (CTCL) and relapsed acute myeloid leukemia (AML), respectively. Such targetable agents, including RFB4(dsFv)-PE38 (BL22), IL13-PE38QQR, and Tf-CRM107, are being tested in clinical trials. Several agents using unique technology such as a cleavable adapter or immunoliposomes with antibodies are also in the preclinical stage. This review summarizes the generation, mechanism, and development of these agents. In addition, possible future directions of this therapeutic approach are discussed

Tea Polyphenols Regulate Key Mediators on Inflammatory Cardiovascular Diseases

Tea polyphenols known as catechins are key components with many biological functions, including anti-inflammatory, antioxidative, and anticarcinogenic effects. These effects are induced by the suppression of several inflammatory factors including nuclear factor-kappa B (NF-κB). While these characteristics of catechins have been well documented, actions of catechins as mediators on inflammation-related cardiovascular diseases have not yet been well investigated. In this article, we reviewed recent papers to reveal the anti-inflammatory effects of catechins in cardiovascular diseases. In our laboratory, we performed oral administration of catechins into murine and rat models of cardiac transplantation, myocarditis, myocardial ischemia, and atherosclerosis to reveal the effects of catechins on the inflammation-induced ventricular and arterial remodeling. From our results, catechins are potent agents for the treatment and prevention of inflammation-related cardiovascular diseases because they are critically involved in the suppression of proinflammatory signaling pathways

Nucleic Acid Drugs for Prevention of Cardiac Rejection

Heart transplantation has been broadly performed in humans. However, occurrence of acute and chronic rejection has not yet been resolved. Several inflammatory factors, such as cytokines and adhesion molecules, enhance the rejection. The graft arterial disease (GAD), which is a type of chronic rejection, is characterized by intimal thickening comprised of proliferative smooth muscle cells. Specific treatments that target the attenuation of acute rejection and GAD formation have not been well studied in cardiac transplantation. Recent progress in the nucleic acid drugs, such as antisense oligodeoxynucleotides (ODNs) to regulate the transcription of disease-related genes, has important roles in therapeutic applications. Transfection of cis-element double-stranded DNA, named as “decoy,” has been also reported to be a useful nucleic acid drug. This decoy strategy has been not only a useful method for the experimental studies of gene regulation but also a novel clinical strategy. In this paper, we reviewed the experimental results of NF-κB, E2F, AP-1, and STAT-1 decoy and other ODNs using the experimental heart transplant models

Hepatocyte growth factor gene therapy reduces ventricular arrhythmia in animal models of myocardial ischemia.

It was recently reported that gene therapy using hepatocyte growth factor (HGF) has the potential to preserve cardiac function after myocardial ischemia. We speculated that this HGF gene therapy could also prevent ventricular arrhythmia. To investigate this possibility, we examined the antiarrhythmic effect of HGF gene therapy in rat acute and old myocardial infarction models. Myocardial ischemia was induced by ligation of the left descending coronary artery. Hemagglutinating virus of Japan (HVJ)-coated liposome containing HGF genes were injected directly into the myocardium fourteen days before programmed pacing. Ventricular fibrillation (VF)was induced by programmed pacing. The VF duration was reduced and the VF threshold increased after HGF gene therapy ( p&#60; 0.01). Histological analyses revealed that the number of vessels in the ischemic border zone was greatly increased after HGF gene injection. These findings revealed that HGF gene therapy has an anti-arrhythmic effect after myocardial ischemia.</p

Inhibitory effect of ribbon-type NF-κB decoy oligodeoxynucleotides on osteoclast induction and activity in vitro and in vivo

In this study we examined the effect of ribbon-type (circular-type) NF-κB decoy oligodeoxynucleotides (RNODN) on osteoclast induction and activity. We extracted bone marrow cells from the femurs of rats and incubated non-adherent cells with receptor activator of nuclear factor κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). First, transfer efficiency into osteoclasts and their precursors, resistance to exonuclease, and binding activity of decoy to NF-κB were examined. Next, to examine the effect of RNODN on osteoclast induction and activity, osteoclast differentiation and pit formation assays were performed. RNODN were injected into the ankle joints of rats with collagen-induced arthritis. Joint destruction and osteoclast activity were examined by histological study. The resistance of RNODN to exonuclease and their binding activity on NF-κB were both greater than those of phosphorothionated NF-κB decoy oligodeoxynucleotides. The absolute number of multinucleate cells scoring positive for tartrate-resistant acid phosphatase was significantly decreased in the RNODN-treated group. The average calcified matrix resorbed area was significantly decreased in the RNODN-treated group. Histological study showed marked suppression of joint destruction and osteoclast activity by intra-articular injection of RNODN. These results suggest the inhibitory effect of RNODN on the induction and activity of osteoclasts. Direct intra-articular injection of RNODN into the joints may be an effective strategy for the treatment of arthritis

Motion and Speed of the Frail Elderly During Evacuation Process

健常者よりも運動能力が劣る要介護者が迅速に避難できるように準備することは，要介護者の命を救うだけでなく，介護スタッフの命を守ることにも繋がる。しかし，現状では要介護者の避難行動速度すら十分に調査されておらず，要介護者の避難を検討する上での情報が不足している。そこで，本研究では要介護者の避難訓練の様子を撮影し，歩行速度をはじめとする避難行動速度を詳細に調査した。その結果，すべての行動が健常者よりも遅いが，階段の上りと車からの降車が特に時間を要することがわかった。さらに，得られた行動速度を入力にして，エージェントモデルで避難訓練をシミュレートしたところ，シミュレーションの方が1割程度早く避難が完了する結果となった。しかしながら，この避難完了時間の違いは，介護スタッフの行動の中の「シミュレーションに考慮されていない行動」に掛かる時間を考慮することでほぼ解消した。While agent simulation has been used to investigate evacuation scenarios, evacuation speed of frail elderly has not been fully studied yet. Therefore, in order to obtain the evacuation speed of the frail elderly we recorded videos of evacuation drills held in a group home with care. We analyzed evacuation steps in detail through the videos and acquired the evacuation speed of the frail elderly. We also simulated the evacuation drills with an agent model using the observed data. In the simulation result, the evacuation completion time was about 10% faster than the measured time of the evacuation drills. The difference in evacuation completion time can be explained considering the time taken for “unplanned behavior” of caregivers such as stopping to think about the next action

Imaging of isotope diffusion using atomic-scale vibrational spectroscopy

The spatial resolutions of even the most sensitive isotope analysis techniques based on light or ion probes are limited to a few hundred nanometres. Although vibration spectroscopy using electron probes has achieved higher spatial resolution, the detection of isotopes at the atomic level has been challenging so far. Here we show the unambiguous isotopic imaging of 12C carbon atoms embedded in 13C graphene and the monitoring of their self-diffusion via atomic level vibrational spectroscopy. We first grow a domain of 12C carbon atoms in a preexisting crack of 13C graphene, which is then annealed at 600C for several hours. Using scanning transmission electron microscopy electron energy loss spectroscopy, we obtain an isotope map that confirms the segregation of 12C atoms that diffused rapidly. The map also indicates that the graphene layer becomes isotopically homogeneous over 100 nanometre regions after 2 hours. Our results demonstrate the high mobility of carbon atoms during growth and annealing via selfdiffusion. This imaging technique can provide a fundamental methodology for nanoisotope engineering and monitoring, which will aid in the creation of isotope labels and tracing at the nanoscale

901-90 In Vivo Genetic Engineering of Cardiac Cells: Intracoronary Administration of Antisense (AS) Oligonucleotides (ODN)

We have previously documented that transfection of antisense ODN by a highly efficient Sendai virus (HVJ)-liposome delivery system can be utilized to modify lesion formation within the peripheral vasculature in vivo. In this study, we defined the feasibility of modifying cardiac cell gene expression via a catheter-based coronary infusion of AS ODN in rabbits. The coronary artery was cannulated via an over-the-wire approach from the carotid artery. Fluorescein (F)-Iabeled ODN were utilized to evaluate the cellular distribution and kinetics of ODN uptake within the myocardium after a single intraluminal bolus of HVJ-liposomes containing ODN. Cellular uptake of F-ODN was primarily localized in the microvasculature and significant staining was also observed in conduit vessels and cardiac myocytes. Immunohistochemical analysis verified prominent localization of F-ODN within the microvascular endothelium. Expression of F-ODN was observed within 10 minutes, peaked at 1 day, and remained evident for up to one week after transfection by the HVJ-liposome method. In contrast, F-ODN infused within liposomes without the viral particle exhibited transient expression that was undetectable within 3 days. These findings indicate that a single intracoronary bolus infusion of ODN within HVJ-liposomes is a reproducible methodology for delivery of AS ODN to targeted cells within the myocardium. Future studies will characterize the feasibility of using this approach to modify cardiac structure and function via regulating myocardial cell gene expression

Promoter Polymorphism of RGS2 Gene Is Associated with Change of Blood Pressure in Subjects with Antihypertensive Treatment: The Azelnidipine and Temocapril in Hypertensive Patients with Type 2 Diabetes Study

We performed a prospective study to examine the genetic effect on the response to a calcium (Ca) channel blocker, azelnidipine and an ACE inhibitor, temocapril treatment in patients with hypertension, as a part of the prior clinical trial, the Azelnidipine and Temocapril in Hypertensive Patients with Type 2 Diabetes Study (ATTEST). Methods and Results. All subjects who gave informed consent for genetic research were divided into two groups: the subjects treated with azelnidipine or temocapril, for 52 weeks. We selected 18 susceptible genes for hypertension and determined their genotypes using TaqMan PCR method. RNA samples were extracted from peripheral blood, and quantitative real time PCR for all genes was performed using TaqMan method. One of the polymorphisms of the RGS2 gene was extracted as being able to influence the effect of these treatments to reduce BP. At eight weeks, BP change showed a significant interaction between the A-638G polymorphism of Regulator of G protein signaling-2 (RGS2) gene and treatment with azelnidipine or temocapril. There was no gene whose expression was associated with BP phenotypes or the polymorphisms of each gene. Conclusions. A-638G polymorphism of the RGS-2 gene could be a predictive factor for therapeutic performance of Ca channel blockers