a-novel-vivo-gene-transfer-technique-vitro-cell-based-assays-for

A Novel <em>in vivo</em> Gene Transfer Technique and <em>in vitro</em> Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

isolation-identification-vascular-endothelial-cells-from-distinct

Isolation and Identification of Vascular Endothelial Cells from Distinct Adipose Depots for Downstream Applications

Vascular endothelial cells lining the wall of the vascular system play important roles in a variety of physiological processes, including vascular tone ...

Shriners Hospitals for Children - Northern California

University of Delaware

The purpose of the present study was to apply computer algorithms to an administrative data set to identify the prevalence of epilepsy, incidence of epilepsy, and epilepsy-related mortality of patients in a managed care organization (MCO).

Estimating prevalence, incidence, and disease-related mortality for patients with epilepsy in managed care organizations.

A newly isolated gene from Ralstonia sp. M1, encoding an esterase, was cloned in Escherichia coli and its nucleotide sequence determined. The 1.6kb insert revealed one complete open reading frame, predicted to encode an esterase (320 aa, 34.1kDa) with a pI of 9.86. EstR contained a putative oxyanion hole H36G37, a conserved pentapeptide G103HSLG107 and a conserved catalytic His265 and Asp237. The EstR sequence shared 64-70 and 44-48% identity with the hydrolases/acyltransferases from Burkholderia strains and from Ralstonia strains, respectively, 44 and 38% identity with the lactone-specific esterase from Pseudomonas fluorescens and Mesorhizobium loti, respectively. The esterase EstR was expressed with a high level of 41mg/g wet cells. The Ni-NTA-purified esterase EstR showed an optimal activity in the temperature range 60-65 degrees C and pH range 7.5-9.0 towards p-nitrophenyl caproate. The enzyme was found to be highly resistant to many organic solvents especially induced by ethanolamine. Metal ions showed slight effect on esterase activity. The inhibitor phenylmethanesulfonyl fluoride inhibited strongly the esterase. Triton X-45 induced the activation of EstR, but other detergents slightly to strongly decreased or completely inhibited. Among tested p-NP esters, caproate was the most preferential substrate of this esterase.

A novel esterase from Ralstonia sp. M1: gene cloning, sequencing, high-level expression and characterization.

Influenza virus infection causes severe respiratory disease such as that due to avian influenza (H5N1). Influenza A viruses proliferate in human epithelial cells, which produce inflammatory cytokines/chemokines as a "cytokine storm" attenuated with the viral nonstructural protein 1 (NS1). Cytokine/chemokine production in A549 epithelial cells infected with influenza A/H1N1 virus (PR-8) or nonstructural protein 1 (NS1) plasmid was examined in vitro. Because tumor necrosis factor-α (TNF-α) and regulated upon activation normal T-cell expressed and secreted (RANTES) are predominantly produced from cells infected with PR-8 virus, the effects of mRNA knockdown of these cytokines were investigated. Small interfering (si)TNF-α down-regulated RANTES expression and secretion of RANTES, interleukin (IL)-8, and monocyte chemotactic protein-1 (MCP-1). In addition, siRANTES suppressed interferon (IFN)-γ expression and secretion of RANTES, IL-8, and MCP-1, suggesting that TNF-α stimulates production of RANTES, IL-8, MCP-1, and IFN-γ, and RANTES also increased IL-8, MCP-1, and IFN-γ. Furthermore, administration of TNF-α promoted increased secretion of RANTES, IL-8, and MCP-1. Administration of RANTES enhanced IL-6, IL-8, and MCP-1 production without PR-8 infection. These results strongly suggest that, as an initial step, TNF-α regulates RANTES production, followed by increase of IL-6, IL-8, and MCP-1 and IFNs concentrations. At a later stage, cells transfected with viral NS1 plasmid showed production of a large amount of IL-8 and MCP-1 in the presence of the H(2)O(2)-myeloperoxidse (MPO) system, suggesting that NS1 of PR-8 may induce a "cytokine storm" from epithelial cells in the presence of an H(2)O(2)-MPO system.

Key role of regulated upon activation normal T-cell expressed and secreted, nonstructural protein1 and myeloperoxidase in cytokine storm induced by influenza virus PR-8 (A/H1N1) infection in A549 bronchial epithelial cells.

A ZnFeO-TiO nanocomposite combining p-type ZnFeO and n-type TiO was successfully fabricated. The ZnFeO-TiO nanocomposite greatly enhanced the bisphenol A (BPA) photodegradation under visible light irradiation at 465 ± 40 nm. Loading TiO with 1 wt% of ZnFeO produced high photocurrent and low charge transfer resistance. The photodegradation rate of BPA by ZnFeO-TiO, which was highly dependent on the water chemistry including pH, anions, and humic acid, was 20.8-21.4 times higher than that of commercial TiO photocatalysts. Chloride and sulfate ions enhanced BPA photodegradation mostly due to the production of more radical species; whereas nitrate, dihydrogen phosphate, and bicarbonate ions decreased the photodegradation rate of BPA due to the scavenge of hydroxyl radicals. The photoactivity and recyclability of ZnFeO-TiO in lake water was also assessed. A near complete BPA removal from lake water was observed under visible light irradiation. Furthermore, >90% of photocatalytic activity toward BPA degradation was achieved in 5 cycles of continuous addition of BPA to the lake water. The BPA degradation intermediates were identified by HPLC/MS/MS and possible reaction pathways were proposed. Results clearly demonstrate the excellent visible-light-sensitive photocatalytic degradation of BPA over ZnFeO-TiO composite which has a great application potential for the decomposition of emerging contaminants in impaired waters.

Photocatalytic degradation of bisphenol A over a ZnFeO/TiO nanocomposite under visible light.

A simple and facile synthesis method is developed for the fabrication of CoO loaded ordered mesoporous carbon nitride (CoO@meso-CN) composites, at various CoO loadings, and used, for the first time, to activate persulfate (PS) for methylene blue (MB) degradation. The interfacial interaction between the ultrafine CoO nanoparticles, immobilized by high surface area, regular mesopores, and graphitic nature of the meso-CN support can further enhance the catalytic activation of PS for methylene blue (MB) degradation. Among all catalysts studied, the 5-wt% CoO@meso-CN exhibits the best catalytic performance with a k of 0.264 min. High initial pH, especially at pH-11, is more beneficial for PS activation. Furthermore, the CoO@meso-CN nanocatalyst is highly stable with a consistently high degree of MB degradation and negligible cobalt leaching for at least 5 consecutive catalytic cycles. Both SO and OH are the major reactive species based on results of EPR and quenching experiments. The degradation intermediates of MB are also identified by HPLC/MS/MS and the possible degradation pathway is proposed. Results clearly demonstrate that CoO@meso-CN is a promising green catalyst with enormous potential for the remediation of hazardous chemicals using PS.

Activation of persulfate by CoO nanoparticles loaded on 3D mesoporous carbon nitride (CoO@meso-CN) for the degradation of methylene blue (MB).

A facile impregnation method was used to fabricate a hybrid CoO-3D ordered mesoporous carbon nitride (CoO@mpgCN) catalyst that effectively activated peroxymonosulfate (PMS) for the degradation of pharmaceutical chemical, exemplified by antibiotic sulfamethoxazole (SMX) in aqueous solutions. The CoO@mpgCN/PMS system exhibited high catalytic reactivity and SMX removal efficiency over a wide pH range with an observed rate constant (k) of 0.314 min. Furthermore, CoO@mpgCN was stable with consistently high degree of SMX degradation without having cobalt dissolution and loss of catalytic activity for at least five consecutive cycles. The significant catalysis performance of CoO@mpgCN was due to its uniformly distributed mesopores, large specific surface area, and high electron transfer ability at the active CoO sites. Both quenching experiments and electron paramagnetic resonance (EPR) analysis verified the yield, in abundance, of highly active species, specifically SO and OH from the CoO@mpgCN activation of PMS, primarily. Hence, SMX degradation followed a radical chain reaction mechanism. The result of this study revealed a novel prospective of CoO@mpgCN composite as PMS activator for the remediation of recalcitrant pollutants in water.

CoO-3D ordered mesoporous carbon nitride (CoO@mpgCN) composite as peroxymonosulfate activator for the degradation of sulfamethoxazole in water.

General lipid-lowering strategies exhibit clinical benefit, however, adverse effects and low adherence of relevant pharmacotherapies warrants the investigation into distinct avenues for preventing dyslipidemia-induced cardiovascular disease. Ion channels play an important role in the maintenance of vascular tone, the impairment of which is a critical precursor to disease progression. Recent evidence suggests that the dysregulation of ion channel function in dyslipidemia is one of many contributors to the advancement of cardiovascular disease thus bringing to light a novel yet putative therapeutic avenue for preventing the progression of disease mechanisms. Increasing evidence suggests that lipid regulation of ion channels often occurs through direct binding of the lipid with the ion channel thereby creating a potential therapeutic target wherein preventing specific lipid-ion channel interactions, perhaps in combination with established lipid lowering therapies, may restore ion channel function and the proper control of vascular tone. Here we first detail specific examples of lipid-ion channel interactions that promote vascular dysfunction and highlight the benefits of preventing such interactions. We next discuss the putative therapeutic avenues, such as peptides, monoclonal antibodies, and aspects of nanomedicine that may be utilized to prevent pathological lipid-ion channel interactions. Finally, we discuss the experimental challenges with identifying lipid-ion channel interactions as well as the likely pitfalls with developing the aforementioned putative strategies.

Thanh Nguyen

Institute for Pediatric Regenerative Medicine

Shriners Hospitals for Children - Northern California

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Isolation and Identification of Vascular Endothelial Cells from Distinct Adipose Depots for Downstream Applications

Thanh Nguyen

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Institute for Pediatric Regenerative Medicine

Shriners Hospitals for Children - Northern California

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Isolation and Identification of Vascular Endothelial Cells from Distinct Adipose Depots for Downstream Applications

Institute for Pediatric Regenerative Medicine