Tumors require ongoing angiogenesis to support their growth. Inhibition of angiogenesis by production of angiostatic factors should be a viable approach for cancer gene therapy. Endostatin, a potent angiostatic factor, was expressed in mouse muscle and secreted into the bloodstream for up to 2 weeks after a single intramuscular administration of the endostatin gene. The biological activity of the expressed endostatin was demonstrated by its ability to inhibit systemic angiogenesis. Moreover, the sustained production of endostatin by intramuscular gene therapy inhibited both the growth of primary tumors and the development of metastatic lesions. These results demonstrate the potential utility of intramuscular delivery of an antiangiogenic gene for treatment of disseminated cancers.

The Agency for Healthcare Research and Quality (AHRQ), through its Evidencebased Practice Centers (EPCs), sponsors the development of evidence reports and technology assessments to assist publicand private-sector organizations in their efforts to improve the quality of health care in the United States. The reports and assessments provide organizations with comprehensive, science-based information on common, costly medical conditions and new health care technologies. The EPCs systematically review the relevant scientific literature on topics assigned to them by AHRQ and conduct additional analyses when appropriate prior to developing their reports and assessments. AHRQ expects that the EPC evidence reports and technology assessments will inform individual health plans, providers, and purchasers as well as the health care system as a whole by providing important information to help improve health care quality. The full report and this summary are available at www.effectivehealthcare.ahrq. gov/reports/final.cfm. d. In patients with chronic pain, what is the comparative effectiveness of opioids plus nonopioid interventions (pharmacological or nonpharmacological) versus opioids or nonopioid interventions alone on outcomes related to pain, function, quality of life, and doses of opioids used? Key Question 2. Harms and Adverse Events a. In patients with chronic pain, what are the risks of opioids versus placebo or no opioid on: (1) opioid abuse, addiction, and related outcomes; (2) overdose; and (3) other harms, including gastrointestinal-related harms, falls, fractures, motor vehicle accidents, endocrinological harms, infections, cardiovascular 3 events, cognitive harms, and psychological harms (e.g., depression)? b. How do harms vary depending on: (1) the specific type or cause of pain (e.g., neuropathic, musculoskeletal [including back pain], fibromyalgia, sickle cell disease, inflammatory pain, headache disorders); (2) patient demographics; (3) patient comorbidities (including past or current substance use disorder or at high risk for addiction); (4) the dose of opioids used? Key Question 3. Dosing Strategies

Stable cationic lipid/DNA complexes were formed by solubilizing cationic liposomes with 1% octylglucoside and complexing a DNA plasmid with the lipid in the presence of detergent. Removal of the detergent by dialysis yielded a lipid/DNA suspension that was able to transfect tissue culture cells up to 90 days after formation with no loss in activity. Similar levels of gene transfer were obtained by mixing the cationic lipid in a liposome form with DNA just prior to cell addition. However, expression was completely lost 24 hr after mixing. The transfection efficiency of the stable complex in 15% fetal calf serum was 30% of that obtained in the absence of serum, whereas the transient complex was completely inactivated with 2% fetal calf serum. A 90-day stability study comparing various storage conditions showed that the stable complex could be stored frozen or as a suspension at 4°C with no loss in transfection efficiency. Centrifugation of the stable complex produced a pellet that contained approximately 90%o of the DNA and 10% of the lipid.Transfection of cells with the resuspended pellet and the supernatant showed that the majority of the transfection activity was in the pellet and all the toxicity was in the supernatant. Formation of a stable cationic lipid/DNA complex has produced a transfection vehicle that can be stored indefinitely, can be concentrated with no loss in transfection efficiency, and the toxicity levels can be greatly reduced when the active complex is isolated from the uncomplexed lipid.The field of gene therapy has produced a diverse spectrum of gene delivery vehicles ranging from replication incompetent viruses to DNA formulated with various delivery vehicles. Administration of DNA alone has yielded successful gene transfer for isolated applications (1). However, the majority of applications requires the assistance of a delivery vehicle to facilitate gene transfer. Gene delivery vehicles have included: cationic lipids (2-6), poly-L-lysine conjugates (7-11), liposomes (12), and polymers (13,14 MATERIALS AND METHODS Materials. Lipofectamine was purchased from GIBCO/ BRL/Life Technologies. Lipofectamine is supplied as a liposome suspension consisting of 2 mg/ml of DOSPA/DOPE (3:1 wt/wt; or 1.53:1 mol/mol). DOTAP liposomes were obtained from Boehringer Mannheim. Plasmid DNA containing the ,3-galactosidase (,B-gal) reporter gene sequence under the CMV promoter (pCMVf3) was purchased from Clontech. ,B-gal was purchased from Boehringer Mannheim.DNA Purification. The plasmids were amplified in E. coli (DH1013 from GIBCO/BRL) and grown in circle grow (Bio 101). Bacteria were lysed and plasmid was purified according to Qiagen (Chatsworth, CA) giga-kit preparation protocol. Care was taken to remove endotoxin (lipopolysaccharide) from the plasmids using polymixin-B columns (Bio-Rad). The DNA was mixed with the polymixin-B resin and agitated at 4°C on a rocker overnight. The DNA was separated from the column support and concentrated by ethanol precipitation with 0.3 M sodium acetate. The DNA was ...

We asked whether the hypoxia-regulated factor, insulin-like growth factor binding protein-3 (IGFBP3), could modulate stem cell factor receptor (c-kit ؉ ), stem cell antigen-1 (sca-1 ؉ ), hematopoietic stem cell (HSC), or CD34 ؉ endothelial precursor cell (EPC) function. Exposure of CD34 ؉ EPCs to IGFBP3 resulted in rapid differentiation into endothelial cells and dose-dependent increases in cell migration and capillary tube formation. IGFBP3-expressing plasmid was injected into the vitreous of neonatal mice undergoing the oxygeninduced retinopathy (OIR) model. In separate studies, GFP-expressing HSCs were transfected with IGFBP3 plasmid and injected into the vitreous of OIR mice. Administering either IGFBP3 plasmid alone or HSCs transfected with the plasmid resulted in a similar reduction in areas of vasoobliteration, protection of the developing vasculature from hyperoxia-induced regression, and reduction in preretinal neovascularization compared to control plasmid or HSCs transfected with control plasmid. In conclusion, IGFBP3 mediates EPC migration, differentiation, and capillary formation in vitro. Targeted expression of IGFBP3 protects the vasculature from damage and promotes proper vascular repair after hyperoxic insult in the OIR model. IGFBP3 expression may represent a physiological adaptation to ischemia and potentially a therapeutic target for treatment of ischemic conditions. IGFBP3 ͉ angiogenesis ͉ retinopathy of prematurity V ascular damage associated with diabetic retinopathy and retinopathy of prematurity (ROP) results from tissue ischemia, and, subsequently, this ischemia leads to development of pathological neovascularization. Insulin-like growth factor 1 (IGF1) is required for normal retinal vascular development because vascular development is arrested in its absence despite the presence of VEGF (1). Development of ROP is associated with low levels of IGF1 (2) because the lack of IGF1 in the early neonatal period leads to the development of avascular retina, which results in ROP (3). However, unregulated IGF1 expression can lead to pathological neovascularization (4-13), and IGF1 receptor (IGF1R) antagonists are able to suppress retinal neovascularization in vivo by inhibiting VEGF signaling (1).The effects of IGF1 are mediated by IGF1R and modulated by complex interactions with IGF binding proteins (IGFBPs), which are also modulated at multiple levels. Six IGFBPs function as transporter proteins and storage pools for IGF1 in a tissue-and developmental stage-specific manner. Phosphorylation, proteolysis, polymerization (8), and cell or matrix association (9) regulates the functions of IGFBPs. Specific IGFBPs have been shown to either stimulate or inhibit IGF1 action (10).IGFBP3, the best studied and most abundant of these binding proteins, carries Ն75% of serum IGF1 and IGF2 in heterotrimeric complexes. Besides its endocrine effects, IGFBP3 has auto-and paracrine actions affecting cell mobility, adhesion, apoptosis, survival, and the cell cycle (14,15). Like the other IGFBPs, IGFBP3 has IGF1-in...

Cities produce considerable ecological light pollution (ELP), yet the effects of artificial night lighting on biological communities and ecosystem function have not been fully explored. From June 2010 to June 2011, we surveyed aquatic emergent insects, riparian arthropods entering the water, and riparian spiders of the family Tetragnathidae at nine stream reaches representing common ambient ELP levels of Columbus, Ohio, USA, streams (low, 0.1-0.5 lux; moderate, 0.6-2.0 lux; high, 2.1-4.0 lux). In August 2011, we experimentally increased light levels at the low- and moderate-treatment reaches to 10-12 lux to represent urban streams exposed to extremely high levels of ELP. Although season exerted the dominant influence on invertebrate fluxes over the course of the year, when analyzed by season, we found that light strongly influenced multiple invertebrate responses. The experimental light addition resulted in a 44% decrease in tetragnathid spider density (P = 0.035), decreases of 16% in family richness (P = 0.040) and 76% in mean body size (P = 0.022) of aquatic emergent insects, and a 309% increase in mean body size of terrestrial arthropods (P = 0.015). Our results provide evidence that artificial light sources can alter community structure and ecosystem function in streams via changes in reciprocal aquatic-terrestrial fluxes of invertebrates.