Reduction of choroidal neovascularization in mice by adeno-associated virus-delivered anti-vascular endothelial growth factor short hairpin RNA.

BACKGROUND: Strategies leading to the long-term suppression of inappropriate ocular angiogenesis are required to avoid the need for repetitive monthly injections for treatment of diseases of the eye, such as age-related macular degeneration (AMD). The present study aimed to develop a strategy for the sustained repression of vascular endothelial growth factor (VEGF), which is identified as the key player in exudative AMD. METHODS: We have employed short hairpin (sh)RNAs combined with adeno-associated virus (AAV) delivery to obtain the targeted expression of potent gene-regulatory molecules. Anti-VEGF shRNAs were analyzed in human retinal pigment epithelial (RPE) cells using Renilla luciferase screening. For in vivo delivery of the most potent shRNA, self-complementary AAV vectors were packaged in serotype 8 capsids (scAAV2/8-hU6-sh9). In vivo efficacy was evaluated either by injection of scAAV2/8-hU6-sh9 into murine hind limb muscles or in a laser-induced murine model of choroidal neovascularization (CNV) following scAAV2/8-hU6-sh9 subretinal delivery. RESULTS: Plasmids encoding anti-VEGF shRNAs showed efficient knockdown of human VEGF in RPEs. Intramuscular administration led to localized expression and 91% knockdown of endogenous murine (m)VEGF. Subsequently, the ability of AAV2/8-encoded shRNAs to impair vessel formation was evaluated in the murine model of CNV. In this model, the sizes of the CNV were significantly reduced (up to 48%) following scAAV2/8-hU6-sh9 subretinal delivery. CONCLUSIONS: Using anti-VEGF vectors, we have demonstrated efficient silencing of endogenous mVEGF and showed that subretinal administration of scAAV2/8-hU6-sh9 has the ability to impair vessel formation in an AMD animal model. Thus, AAV-encoded shRNA can be used for the inhibition of neovascularization, leading to the development of sustained anti-VEGF therapy. Copyright © 2012 John Wiley & Sons, Ltd.

Dairy calcium supplementation in overweight or obese persons: its effect on markers of fat metabolism

BACKGROUND: Dairy calcium supplementation has been proposed to increase fat oxidation and to inhibit lipogenesis. OBJECTIVE: We aimed to investigate the effects of calcium supplementation on markers of fat metabolism. DESIGN: In a placebo-controlled, crossover experiment, 10 overweight or obese subjects who were low calcium consumers received 800 mg dairy Ca/d for 5 wk. After 4 wk, adipose tissue was taken for biopsy for analysis of gene expression. Respiratory exchange, glycerol turnover, and subcutaneous adipose tissue microdialysis were performed for 7 h after consumption of 400 mg Ca or placebo, and the ingestion of either randomized slow-release caffeine (SRC; 300 mg) or lactose (500 mg). One week later, the test was repeated with the SRC or lactose crossover. RESULTS: Calcium supplementation increased urinary calcium excretion by 16% (P = 0.017) but did not alter plasma parathyroid hormone or osteocalcin concentrations. Resting energy expenditure (59.9 +/- 3.0 or 59.6 +/- 3.3 kcal/h), fat oxidation (58.4 +/- 2.5 or 53.8 +/- 2.2 mg/min), plasma free fatty acid concentrations (0.63 +/- 0.02 or 0.62 +/- 0.03 mmol/L), and glycerol turnover (3.63 +/- 0.41 or 3.70 +/- 0.38 micromol . kg(-1) . min(-1)) were similar with or without calcium, respectively. SRC significantly increased free fatty acid concentrations, resting fat oxidation, and resting energy expenditure. During microdialysis, epinephrine increased dialysate glycerol concentrations by 250% without and 254% with calcium. Expression of 7 key metabolic genes in subcutaneous adipose tissue was not affected by calcium supplementation. CONCLUSION: Dairy calcium supplementation in overweight subjects with habitually low calcium intakes failed to alter fat metabolism and energy expenditure under resting conditions and during acute stimulation by caffeine or epinephrine

Oxygen isotope sector zoning in natural hydrothermal quartz

Oxygen isotope measurements using SIMS and laser-fluorination methods confirm the presence of concentric and sector zoning in low-temperature (200 degrees C to < 400 degrees C) hydrothermal quartz from Alpine veins. While concentric zoning is most readily explained by changes in the chemical composition of the fluid or temperature of crystallization, the reasons for sector zoning are more difficult to explain. Relative enrichment in (18)O for crystallographically different sectors of quartz corresponds to m > r > z. Sector zoning is, however, largely limited to the exterior zones of crystals and/or to crystals with large Al (> 1000 ppm) and trace element contents, probably formed at temperatures < 250 degrees C. Differences in delta(18)O between the prismatic (m) relative to the rhombohedral (r and z) growth sectors of up to 2 parts per thousand can be explained by a combination of a face-related crystallographic and/or a growth rate control. In contrast, isotopic sector zoning of up to about 1.5 parts per thousand amongst the different rhombohedral faces increases in parallel with the trace element content and is likely to represent disequilibrium growth. This is indicated by non-systematic, up to 2 parts per thousand, differences within single growth zones and the irregular, larger or smaller, delta(18)O values (of several permil) of the exterior compared to the inner zones of the same crystals. Disequilibrium growth may be related to the large trace element content incorporated into the growing quartz at lower temperatures (< 250 degrees C) and/or be related to fluid-vapour separation, allowing crystal growth from both a vapour as well as a liquid phase.

Issue review : impact on the insurance premium tax rate reduction .

In 2002, Senate File 2318, Insurance Premium Tax Reduction Act, the Iowa General Assembly approved a reduction in the insurance premium tax rate from 2 percent to 1 percent. The reduction was phased in at the rate of 1/4 percent annual increments over a five-year period, beginning with life and health insurance policy payments made on or after January 1st, 2003. Calendar year 2007 was the first year all premiums were taxed at 1 percent and fiscal year 2008 was the first full fiscal year at the 1 percent rate. Insurance premiums tax is paid only by insurance companies. The companies that benefit from the tax reduction are uniquely identified in government employment statistics. This allows for a unique opportunity to evaluate both the revenue impact and the employment impact of the tax rate reduction in Senate File 2318. This issue review focuses on the General Fund revenue impact of the rate reduction and the Iowa employment trends for the insurance industry since the rate reduction legislation was approved.

Mixing of Rhône River water in Lake Geneva (Switzerland-France) inferred from stable hydrogen and oxygen isotope profiles

Depth profiles were sampled at different locations throughout Lake Geneva on a monthly and seasonal basis over the course of 2 years and analysed for their stable hydrogen and oxygen isotope compositions. The isotopic compositions indicate an isotopic stratification in the metalimnion during summer and fall. This is related to mixing of Rhône River water, which in summer is dominated by snow and glacier melt waters, and lake water, with the latter having a homogenous isotopic composition. The observed interflow layer is 7-15 m thick and can be traced by the distinct stable isotope composition of the water for about 55 km throughout the lake as well as into shallow bay regions. Depth of the interflow layer close to the Rhône River mouth is similar to those previously described based on echo-soundings and turbidity profiles of sediment dispersion. In contrast to previous descriptions of the interflow within Lake Geneva, the stable isotope compositions allow for direct, natural tracing of the Rhône River water even in cases where the turbidity and conductivity measurements do not indicate such an interflow. In addition, the method allows for a quantification of the Rhône River and lake water in the interflow with the fraction of Rhône River water within the interflow estimated to be up to 37% in summer. The isotopic composition further indicates different vertical mixing processes within the two lake basins of Lake Geneva, related to the density gradients and local stability within the water column. The method may be applicable to other lakes in catchments with large differences in the topography as water that originates from high altitudes or glaciers has a distinct oxygen and hydrogen isotope composition compared to other sources of water originating at lower altitudes and/or from direct precipitation over the lake. Stable isotope measurements thus improve the understanding of the circulation of water within the lake, which is fundamental for an evaluation of the water residence times, dissolved pollutant and nutrient transport as well as oxygenation.

FAM-MDR: a flexible family-based multifactor dimensionality reduction technique to detect epistasis using related individuals

We propose a novel multifactor dimensionality reduction method for epistasis detection in small or extended pedigrees, FAM-MDR. It combines features of the Genome-wide Rapid Association using Mixed Model And Regression approach (GRAMMAR) with Model-Based MDR (MB-MDR). We focus on continuous traits, although the method is general and can be used for outcomes of any type, including binary and censored traits. When comparing FAM-MDR with Pedigree-based Generalized MDR (PGMDR), which is a generalization of Multifactor Dimensionality Reduction (MDR) to continuous traits and related individuals, FAM-MDR was found to outperform PGMDR in terms of power, in most of the considered simulated scenarios. Additional simulations revealed that PGMDR does not appropriately deal with multiple testing and consequently gives rise to overly optimistic results. FAM-MDR adequately deals with multiple testing in epistasis screens and is in contrast rather conservative, by construction. Furthermore, simulations show that correcting for lower order (main) effects is of utmost importance when claiming epistasis. As Type 2 Diabetes Mellitus (T2DM) is a complex phenotype likely influenced by gene-gene interactions, we applied FAM-MDR to examine data on glucose area-under-the-curve (GAUC), an endophenotype of T2DM for which multiple independent genetic associations have been observed, in the Amish Family Diabetes Study (AFDS). This application reveals that FAM-MDR makes more efficient use of the available data than PGMDR and can deal with multi-generational pedigrees more easily. In conclusion, we have validated FAM-MDR and compared it to PGMDR, the current state-of-the-art MDR method for family data, using both simulations and a practical dataset. FAM-MDR is found to outperform PGMDR in that it handles the multiple testing issue more correctly, has increased power, and efficiently uses all available information.

Reduction of ARNT in myeloid cells causes immune suppression and delayed wound healing.

Aryl hydrocarbon receptor nuclear translocator (ARNT) is a transcription factor that binds to partners to mediate responses to environmental signals. To investigate its role in the innate immune system, floxed ARNT mice were bred with lysozyme M-Cre recombinase animals to generate lysozyme M-ARNT (LAR) mice with reduced ARNT expression. Myeloid cells of LAR mice had altered mRNA expression and delayed wound healing. Interestingly, when the animals were rendered diabetic, the difference in wound healing between the LAR mice and their littermate controls was no longer present, suggesting that decreased myeloid cell ARNT function may be an important factor in impaired wound healing in diabetes. Deferoxamine (DFO) improves wound healing by increasing hypoxia-inducible factors, which require ARNT for function. DFO was not effective in wounds of LAR mice, again suggesting that myeloid cells are important for normal wound healing and for the full benefit of DFO. These findings suggest that myeloid ARNT is important for immune function and wound healing. Increasing ARNT and, more specifically, myeloid ARNT may be a therapeutic strategy to improve wound healing.

Effect of transnasal insufflation on sleep disordered breathing in acute stroke: a preliminary study.

BACKGROUND AND PURPOSE: Sleep disordered breathing (SDB) is frequent in acute stroke patients and is associated with early neurologic worsening and poor outcome. Although continuous positive airway pressure (CPAP) effectively treats SDB, compliance is low. The objective of the present study was to assess the tolerance and the efficacy of a continuous high-flow-rate air administered through an open nasal cannula (transnasal insufflation, TNI), a less-intrusive method, to treat SDB in acute stroke patients. METHODS: Ten patients (age, 56.8 ± 10.7 years), with SDB ranging from moderate to severe (apnea-hypopnea index, AHI, >15/h of sleep) and on a standard sleep study at a mean of 4.8 ± 3.7 days after ischemic stroke (range, 1-15 days), were selected. The night after, they underwent a second sleep study while receiving TNI (18 L/min). RESULTS: TNI was well tolerated by all patients. For the entire group, TNI decreased the AHI from 40.4 ± 25.7 to 30.8 ± 25.7/h (p = 0.001) and the oxygen desaturation index >3% from 40.7 ± 28.4 to 31 ± 22.5/h (p = 0.02). All participants except one showed a decrease in AHI. The percentage of slow-wave sleep significantly increased with TNI from 16.7 ± 8.2% to 22.3 ± 7.4% (p = 0.01). There was also a trend toward a reduction in markers of sleep disruption (number of awakenings, arousal index). CONCLUSIONS: TNI improves SDB indices, and possibly sleep parameters, in stroke patients. Although these changes are modest, our findings suggest that TNI is a viable treatment alternative to CPAP in patients with SDB in the acute phase of ischemic stroke.

Carbon and oxygen isotope working standards from C-3 and C-4 photosynthates

A preparation of organic working standards for the online measurement of C-13/C-12 and O-18/O-16 ratios in biological material is presented. The organic working standards are simple and inexpensive C-3 and C-4 carbohydrates ( sugars or cellulose) from distinct geographic origin, including white sugar, toilet and XEROX papers from Switzerland, maize from Ivory Coast, cane sugar from Brazil, papyrus from Egypt, and the core of the stem of a Cyperus papyrus plant from Kenya. These photosynthetic products were compared with International Atomic Energy standards CH-3 and CH-6 and other calibration materials. The presented working standards cover a 15 parts per thousand range of C-13/C-12 ratios and 9 parts per thousand for O-18/O-16, with a precision < +/- 0.2 parts per thousand for n > 10.

Asphalt Stabilization (Asphadur)/Reduction of Reflection Cracks (Monsanto Bidim Synthetic Fabric), HR-511, 1979

ASPHALT STABILIZATION (ASPHADUR): Asphadur (now called 3M Additive 5990) was incorporated into asphaltic concrete on a lane delineation, AC resurfacing, project in Council Bluffs. The experimental feature was included in the eastbound lanes of Interstate 480, beginning at the bridge over the Missouri River and ending at the bridge over North 41st Street. The project was constructed in October 1979. The objective of the project was to investigate the manufacturer's claims of improved strength, stability and durability of an asphalt mix. REDUCTION OF REFLECTION CRACKS (MONSANTO BIDIM SYNTHETIC FABRIC): A lane delineation project was constructed in the eastbound lanes of Interstate 480 in Council Bluffs. A synthetic fabric, Monsanto Bidim C-28, was placed between the portland cement concrete and two inches of Type A asphaltic concrete resurfacing containing Asphadur. The experimental feature began at the bridge over the Missouri River and ended at the bridge over North 41st Street. The project was constructed in October 1979. The objective of this experimental project was to determine the effectiveness of the fabric in reducing reflective cracking in an asphaltic concrete overlay.

Reduction of Concrete Deterioration by Ettringite Using Crystal Growth Inhibition Techniques: Part II Field Evaluation of Inhibitor Effectiveness, TR-469, 2004

The effects of diethylenetriaminpenta(methylenephosphonic acid) (DTPMP), a phosphonate inhibitor, on the growth of delayed ettringite have been evaluated using concrete in highway US 20 near Williams, Iowa, and the cores of six highways subject to moderate (built in 1992) or minor (built in 1997) deterioration. Application of 0.01 and 0.1 vol. % DTPMP to cores was made on a weekly or monthly basis for one year under controlled laboratory-based freeze-thaw and wet-dry conditions over a temperature range of -15 degrees to 58 degrees C to mimic extremes in Iowa roadway conditions. The same concentrations of phosphonate were also applied to cores left outside (roof of Science I at Iowa State University) over the same period of time. Nineteen applications of 0.1 vol. % DTPMP with added deicing salt solution (about 23 weight % NACL) were made to US 20 during the winters of 2003 and 2004. In untreated samples, air voids, pores, and occasional cracks are lined with acicular ettringite crystals (up to 50 micrometers in length) whereas air voids, pores, and cracks in concrete from the westbound lane of US 20 are devoid of ettringite up to a depth of about 0.5 mm from the surface of the concrete. Ettringite is also absent in zones up to 6 mm from the surface of concrete slabs placed on the roof of Science I and cores subject to laboratory-based freeze-thaw experiments. In these zones, the relatively high concentration of DTPMP caused it to behave as a chelator. Stunted ettringite crystals 5 to 25 micrometers in length, occasionally coated with porlandite, form on the margins of these zones indicating that in these areas DTPMP behaved as an inhibitor due to a reduction in the concentration of phosphonate. Analyses of mixes of ettringite and DTPMP using electrospray mass spectrometry suggests that the stunting of ettringite growth is caused by the adsorption of a Ca2+ ion and a water molecule to deprotonated DTPMP on the surface of the {0001} face of ettringite. It is anticipated that by using a DTPMP concentration of between 0.001 and 0.01 vol. % for the extended life of a highway (i.e. >20 years), deterioration caused by the expansive growth of ettringite will be markedly reduced.

Investigation of Materials for the Reduction and Prevention of Corrosion on Highway Maintenance Equipment, TR-472, 2009

The issue of corrosion of winter maintenance equipment is becoming of greater concern because of the increased use of liquid solutions of ice control chemicals, as opposed to their application in solid form. Being in liquid form, the ice control chemicals can more easily penetrate into the nooks and crannies on equipment and avoid being cleansed from the vehicle. Given this enhanced corrosive ability, methods must be found to minimize corrosion. The methods may include coatings, additives, cleansing techniques, other methods, and may also include doing nothing, and accepting a reduced equipment lifetime as a valid (perhaps) trade off with the enhanced benefits of using liquid ice control chemicals. In reality, some combination of these methods may prove to be optimal. Whatever solutions are selected, they must be relatively cheap and durable. The latter point is critical because of the environment in which maintenance trucks operate, in which scrapes, scratches and dents are facts of life. Protection methods that are not robust simply will not work. The purpose of this study is to determine how corrosion occurs on maintenance trucks, to find methods that would minimize the major corrosion mechanisms, and to

The ADVANCE trial: clarifying the role of perindopril/indapamide fixed-dose combination in the reduction of cardiovascular and renal events in patients with diabetes mellitus.

Patients with type 2 diabetes mellitus exhibit a marked increase in cardiovascular and renal risk. A number of interventional trials have shown that these patients benefit greatly from aggressive BP lowering, especially when the drug regimen comprises an inhibitor of the renin-angiotensin system. The results of the placebo-controlled ADVANCE (Action in Diabetes and Vascular disease: PreterAx and DiamicroN MR Controlled Evaluation) trial, conducted in patients with type 2 diabetes, are exemplary in this respect. The systematic use of a fixed-dose combination containing the ACE inhibitor perindopril and the diuretic indapamide afforded substantial protection against cardiovascular mortality and myocardial infarction, while providing important renoprotection, reducing the development of micro- and macroalbuminuria, and allowing regression of nephropathy. The beneficial effects were obtained regardless of baseline BP and whether or not the patients were receiving antihypertensive therapy.