eNOS transfection of adipose-derived stem cells yields bioactive nitric oxide production and improved results in vascular tissue engineering.

This study evaluates the durability of a novel tissue engineered blood vessel (TEBV) created by seeding a natural vascular tissue scaffold (decellularized human saphenous vein allograft) with autologous adipose-derived stem cells (ASC) differentiated into endothelial-like cells. Previous work with this model revealed the graft to be thrombogenic, likely due to inadequate endothelial differentiation as evidenced by minimal production of nitric oxide (NO). To evaluate the importance of NO expression by the seeded cells, we created TEBV using autologous ASC transfected with the endothelial nitric oxide synthase (eNOS) gene to produce NO. We found that transfected ASC produced NO at levels similar to endothelial cell (EC) controls in vitro which was capable of causing vasorelaxation of aortic specimens ex vivo. TEBV (n = 5) created with NO-producing ASC and implanted as interposition grafts within the aorta of rabbits remained patent for two months and demonstrated a non-thrombogenic surface compared to unseeded controls (n = 5). Despite the xenograft nature of the scaffold, the TEBV structure remained well preserved in seeded grafts. In sum, this study demonstrates that upregulation of NO expression within adult stem cells differentiated towards an endothelial-like lineage imparts a non-thrombogenic phenotype and highlights the importance of NO production by cells to be used as endothelial cell substitutes in vascular tissue engineering applications

Sphingosine-1-phosphate promotes the differentiation of adipose-derived stem cells into endothelial nitric oxide synthase (eNOS) expressing endothelial-like cells.

BACKGROUND: Adipose tissue provides a readily available source of autologous stem cells. Adipose-derived stem cells (ASCs) have been proposed as a source for endothelial cell substitutes for lining the luminal surface of tissue engineered bypass grafts. Endothelial nitric oxide synthase (eNOS) is a key protein in endothelial cell function. Currently, endothelial differentiation from ASCs is limited by poor eNOS expression. The goal of this study was to investigate the role of three molecules, sphingosine-1-phosphate (S1P), bradykinin, and prostaglandin-E1 (PGE1) in ASC endothelial differentiation. Endothelial differentiation markers (CD31, vWF and eNOS) were used to evaluate the level of ASCs differentiation capability. RESULTS: ASCs demonstrated differentiation capability toward to adipose, osteocyte and endothelial like cell phenotypes. Bradykinin, S1P and PGE were used to promote differentiation of ASCs to an endothelial phenotype. Real-time PCR showed that all three molecules induced significantly greater expression of endothelial differentiation markers CD31, vWF and eNOS than untreated cells. Among the three molecules, S1P showed the highest up-regulation on endothelial differentiation markers. Immunostaining confirmed presence of more eNOS in cells treated with S1P than the other groups. Cell growth measurements by MTT assay, cell counting and EdU DNA incorporation suggest that S1P promotes cell growth during ASCs endothelial differentiation. The S1P1 receptor was expressed in ASC-differentiated endothelial cells and S1P induced up-regulation of PI3K. CONCLUSIONS: S1P up-regulates endothelial cell markers including eNOS in ASCs differentiated to endothelial like cells. This up-regulation appears to be mediated by the up-regulation of PI3K via S1P1 receptor. ASCs treated with S1P offer promising use as endothelial cell substitutes for tissue engineered vascular grafts and vascular networks

Perioperative antibiotics for prevention of acute endophthalmitis after cataract surgery

Endophthalmitis is a severe inflammation of the anterior or posterior (or both) chambers of the eye that may be sterile or associated with infection. It is a potentially vision-threatening complication of cataract surgery. Prophylactic measures for endophthalmitis are targeted against various sources of infection

Comparison of vertical cup-to-disc ratio estimates using stereoscopic and monoscopic cameras

Background: The use of monoscopic cameras for glaucoma screening is increasing due to their portability, lower cost, and non-mydriatic capabilities. However, it is important to compare the accuracy of such devices with stereoscopic cameras that are used clinically and are considered the gold standard in optic disc assessment. The aim of this study is to compare vertical cup-to-disc ratio (VCDR) estimates obtained using images taken with a monoscopic and stereoscopic camera. Methods: Participants were selected from the Tema Eye Survey. Eligible subjects had images of at least one eye taken with two cameras. They were classified as meeting the glaucoma threshold if an eye had a VCDR estimate >97.5th percentile, corresponding to >0.725 for this population. Hence, we used 0.725 as the cutoff to group eyes into two categories: positive and negative. We calculated sensitivity, specificity, and predictive values of VCDR assessed by expert readers at a reading center for monoscopic photos using stereoscopic photos as the gold standard. Results: Three hundred and seventy-nine eyes of 206 participants were included in the study. Most participants were female (60.2%) and the most common age group was 50–59 years (36.4%). Sixteen eyes met the glaucoma threshold (VCDR > 0.725). Of these, the VCDR estimates of 14 eyes (87.5%) disagreed on the glaucoma threshold from the two cameras. The sensitivity to detect glaucoma with the monoscopic camera was 14.3% (95% CI: 4.0, 40.3). Conclusions: The low sensitivity of monoscopic photos suggests that stereoscopic photos are more useful in the diagnosis of glaucoma

Plasma Dynamics

Contains reports on three research projects.National Science Foundation (Grant G-9330)United States Air Force, Air Force Cambridge Research Center (Contract AF19(604)-5992)United States ArmyUnited States Air Force (Contract AF19(604)-7400)Lincoln Laboratory (Purchase Order B-00306)Flight Accessories Laboratory, Wright-Patterson Air Force Base (WADD Contract AF33(616)-7624)United States Air Force, Air Force Cambridge Research Laboratories (Contract AF19(604)-4551)United States NavyUnited States Atomic Energy Commission (Contract AT(30-1)-1842

Improving local health through community health workers in Cambodia: challenges and solutions

Volunteer community health workers (CHWs) are an important link between the public health system and the community. The ‘Community Participation Policy for Health’ in Cambodia identifies CHWs as key to local health promotion and as a critical link between district health centres and the community. However, research on the challenges CHWs face and identifying what is required to optimise their performance is limited in the Cambodian context. This research explores the views of CHWs in rural Cambodia, on the challenges they face when implementing health initiatives

The Path to Sustainable Nuclear Energy. Basic and Applied Research Opportunities for Advanced Fuel Cycles

The objective of this report is to identify new basic science that will be the foundation for advances in nuclear fuel-cycle technology in the near term, and for changing the nature of fuel cycles and of the nuclear energy industry in the long term. The goals are to enhance the development of nuclear energy, to maximize energy production in nuclear reactor parks, and to minimize radioactive wastes, other environmental impacts, and proliferation risks. The limitations of the once-through fuel cycle can be overcome by adopting a closed fuel cycle, in which the irradiated fuel is reprocessed and its components are separated into streams that are recycled into a reactor or disposed of in appropriate waste forms. The recycled fuel is irradiated in a reactor, where certain constituents are partially transmuted into heavier isotopes via neutron capture or into lighter isotopes via fission. Fast reactors are required to complete the transmutation of long-lived isotopes. Closed fuel cycles are encompassed by the Department of Energy?s Advanced Fuel Cycle Initiative (AFCI), to which basic scientific research can contribute. Two nuclear reactor system architectures can meet the AFCI objectives: a ?single-tier? system or a ?dual-tier? system. Both begin with light water reactors and incorporate fast reactors. The ?dual-tier? systems transmute some plutonium and neptunium in light water reactors and all remaining transuranic elements (TRUs) in a closed-cycle fast reactor. Basic science initiatives are needed in two broad areas: ? Near-term impacts that can enhance the development of either ?single-tier? or ?dual-tier? AFCI systems, primarily within the next 20 years, through basic research. Examples: Dissolution of spent fuel, separations of elements for TRU recycling and transmutation Design, synthesis, and testing of inert matrix nuclear fuels and non-oxide fuels Invention and development of accurate on-line monitoring systems for chemical and nuclear species in the nuclear fuel cycle Development of advanced tools for designing reactors with reduced margins and lower costs ? Long-term nuclear reactor development requires basic science breakthroughs: Understanding of materials behavior under extreme environmental conditions Creation of new, efficient, environmentally benign chemical separations methods Modeling and simulation to improve nuclear reaction cross-section data, design new materials and separation system, and propagate uncertainties within the fuel cycle Improvement of proliferation resistance by strengthening safeguards technologies and decreasing the attractiveness of nuclear materials A series of translational tools is proposed to advance the AFCI objectives and to bring the basic science concepts and processes promptly into the technological sphere. These tools have the potential to revolutionize the approach to nuclear engineering R&D by replacing lengthy experimental campaigns with a rigorous approach based on modeling, key fundamental experiments, and advanced simulations

Triglyceride Blisters in Lipid Bilayers: Implications for Lipid Droplet Biogenesis and the Mobile Lipid Signal in Cancer Cell Membranes

Triglycerides have a limited solubility, around 3%, in phosphatidylcholine lipid bilayers. Using millisecond-scale course grained molecular dynamics simulations, we show that the model lipid bilayer can accommodate a higher concentration of triolein (TO) than earlier anticipated, by sequestering triolein molecules to the bilayer center in the form of a disordered, isotropic, mobile neutral lipid aggregate, at least 17 nm in diameter, which forms spontaneously, and remains stable on at least the microsecond time scale. The results give credence to the hotly debated existence of mobile neutral lipid aggregates of unknown function present in malignant cells, and to the early biogenesis of lipid droplets accommodated between the two leaflets of the endoplasmic reticulum membrane. The TO aggregates give the bilayer a blister-like appearance, and will hinder the formation of multi-lamellar phases in model, and possibly living membranes. The blisters will result in anomalous membrane probe partitioning, which should be accounted for in the interpretation of probe-related measurements

Modulation of calcification of vascular smooth muscle cells in culture by calcium antagonists, statins, and their combination

Background Vascular calcification is an organized process in which vascular smooth muscle cells (VSMCs) are implicated primarily. The purpose of the present study was to assess the effects of calcium antagonists and statins on VSMC calcification in vitro. Methods VSMC calcification was stimulated by incubation in growth medium supplemented with 10 mmol/l β-glycerophosphate, 8 mmol/l CaCl2, 10 mmol/l sodium pyruvate, 1 μmol/l insulin, 50 μg/ml ascorbic acid, and 100 nmol/l dexamethasone (calcification medium). Calcification, proliferation, and apoptosis of VSMCs were quantified. Results Calcium deposition was stimulated dose-dependently by β-glycerophosphate, CaCl2, and ascorbic acid (all P < 0.01). Addition of amlodipine (0.01–1 μmol/l) to the calcification medium did not affect VSMC calcification. However, atorvastatin (2–50 μmol/l) stimulated calcium deposition dose-dependently. Combining treatments stimulated calcification to a degree similar to that observed with atorvastatin alone. Both atorvastatin and amlodipine inhibited VSMC proliferation at the highest concentration used. Only atorvastatin (50 μmol/l) induced considerable apoptosis of VSMCs. Conclusion In vitro calcification of VSMCs is not affected by amlodipine, but is stimulated by atorvastatin at concentrations ≥10 μmol/l, which could contribute to the plaque-stabilizing effect reported for statins

A median fin derived from the lateral plate mesoderm and the origin of paired fins

The development of paired appendages was a key innovation during evolution and facilitated the aquatic to terrestrial transition of vertebrates. Largely derived from the lateral plate mesoderm (LPM), one hypothesis for the evolution of paired fins invokes derivation from unpaired median fins via a pair of lateral fin folds located between pectoral and pelvic fin territories1. Whilst unpaired and paired fins exhibit similar structural and molecular characteristics, no definitive evidence exists for paired lateral fin folds in larvae or adults of any extant or extinct species. As unpaired fin core components are regarded as exclusively derived from paraxial mesoderm, any transition presumes both co-option of a fin developmental programme to the LPM and bilateral duplication2. Here, we identify that the larval zebrafish unpaired pre-anal fin fold (PAFF) is derived from the LPM and thus may represent a developmental intermediate between median and paired fins. We trace the contribution of LPM to the PAFF in both cyclostomes and gnathostomes, supporting the notion that this is an ancient trait of vertebrates. Finally, we observe that the PAFF can be bifurcated by increasing bone morphogenetic protein signalling, generating LPM-derived paired fin folds. Our work provides evidence that lateral fin folds may have existed as embryonic anlage for elaboration to paired fins