Fluxes of Atmospheric Greenhouse-Gases in Maryland (FLAGG-MD): Emissions of Carbon Dioxide in the Baltimore, MD-Washington, D.C. area.

To study emissions of CO2 in the Baltimore, MD-Washington, D.C. (Balt-Wash) area, an aircraft campaign was conducted in February 2015, as part of the FLAGG-MD (Fluxes of Atmospheric Greenhouse-Gases in Maryland) project. During the campaign, elevated mole fractions of CO2 were observed downwind of the urban center and local power plants. Upwind flight data and HYSPLIT (Hybrid Single Particle Lagrangian Integrated Trajectory) model analyses help account for the impact of emissions outside the Balt-Wash area. The accuracy, precision, and sensitivity of CO2 emissions estimates based on the mass balance approach were assessed for both power plants and cities. Our estimates of CO2 emissions from two local power plants agree well with their CEMS (Continuous Emissions Monitoring Systems) records. For the 16 power plant plumes captured by the aircraft, the mean percentage difference of CO2 emissions was -0.3 %. For the Balt-Wash area as a whole, the 1σ CO2 emission rate uncertainty for any individual aircraft-based mass balance approach experiment was ±38 %. Treating the mass balance experiments, which were repeated seven times within nine days, as individual quantifications of the Balt-Wash CO2 emissions, the estimation uncertainty was ±16 % (standard error of the mean at 95% CL). Our aircraft-based estimate was compared to various bottom-up fossil fuel CO2 (FFCO2) emission inventories. Based on the FLAGG-MD aircraft observations, we estimate 1.9±0.3 MtC of FFCO2 from the Balt-Wash area during the month of February 2015. The mean estimate of FFCO2 from the four bottom-up models was 2.2±0.3 MtC.

Surgical and clinical confirmation of temporal bone CT findings in patients with otosclerosis with failed stapes surgery.

BACKGROUND AND PURPOSE: Prior descriptions of imaging after failed stapes procedures for otosclerosis predated currently available CT technology and/or failed to assess commonly used metallic implants. The purpose of this study was to correlate temporal bone CT findings with clinically and intraoperatively determined causes of surgical failure. MATERIALS AND METHODS: All patients with otosclerosis undergoing stapedectomy between December 1999 and December 2010 were identified from a search of neurotology clinical records. Patients presenting because of failed stapes surgery and having temporal bone CT scans at the time of revision surgery or clinical evaluation were included. Imaging and clinical records were retrospectively evaluated by a medical student, radiology resident, and senior neuroradiologist. Stapes prosthesis complications and relevant anatomic CT findings were correlated to clinical and intraoperative findings. RESULTS: Twenty-two of 340 patients met inclusion criteria. Temporal bone CT findings were correlated to intraoperative findings in 17 of 22 patients and to clinical findings in 5 of 22 patients. Surgically confirmed abnormalities included 7 of 7 incus erosions, 3 of 6 piston re-sizings, 3 of 5 granulation tissues, 3 of 5 prosthesis disconnections, 3 of 4 obliterative otosclerosis, 2 of 2 oval window dislocations, and 1 labyrinthine ossificans. Clinically confirmed abnormalities included 2 cases each of superior semicircular canal dehiscence, and wrong piston size, and 1 each of piston disconnection, labyrinthine ossificans, and intravestibular footplate. CONCLUSIONS: CT evaluation in the setting of failed stapes surgery is challenging. Many postoperative complications such as piston migration, incus necrosis, and overt vestibular penetration are well recognized on temporal bone CT. Of particular note, superior semicircular canal dehiscence is an important contraindication to stapes surgery.

Fluoroscopic and CT fistulography of the first branchial cleft.

We present an unusual case of a complete first branchial cleft fistula communicating between the external auditory canal and the skin near the angle of the mandible. CT and fluoroscopic fistulography were used to establish the presence and course of the tract and to assist in surgical planning. The embryology and classification of first branchial cleft anomalies are discussed, with emphasis on the impact of imaging.

The effect of surface coverage and conformation of poly(ethylene oxide) (PEO) chains of poloxamer 407 on the biological fate of model colloidal drug carriers.

Poloxamer 407 was adsorbed onto the surface of model colloidal drug carriers, polystyrene nanoparticles of 40, 70 and 137 nm in diameter, and the effect of the degree of surface coverage and the conformation of the poly(ethylene oxide) (PEO) chains on biological fate was studied. The relationship between the physicochemical and the biological properties of the nanoparticle systems was also investigated. The adsorbed layer of poloxamer 407 was characterised in terms of percentage surface coverage, thickness of the adsorbed layer and average surface area per PEO chain. Computer modelling of the adsorbed layer was performed (applying the self-consistent field technique), to obtain the structural information of the PEO chains in the layer. The in vitro interaction of the nanoparticles with different degrees of poloxamer 407 surface coverage with serum components and the in vivo biodistribution in the rat model were assessed. The results demonstrated that an increase in the surface coverage with poloxamer 407 resulted in an increased volume fraction of the PEO in the adsorbed layer, further extension of the PEO chains from the surface and closer packing of the chains at the surface. With regard to the interaction with the serum components, an increased surface coverage resulted in a reduction of the amount of serum proteins adsorbed, and, importantly, affected the type of proteins adsorbed. High molecular weight proteins were not adsorbed onto the nanoparticles with a surface coverage above approx. 25%. Following the intravenous administration to rats, even the nanoparticles with the lowest degree of surface coverage (approx. 5%) showed improved circulation profiles relative to the uncoated nanoparticles. The effect was more pronounced for the 40 nm nanoparticles. A further increase in the surface coverage to approx. 25% resulted in a significant increase in circulation time, as compared to uncoated and 5% coated systems, for all sizes of nanoparticles. Importantly, it was found that a long in vivo blood circulation time could be achieved for nanoparticles with a relatively low degree of surface coverage with PEO chains.

Development of systems for targeting the regional lymph nodes for diagnostic imaging: in vivo behaviour of colloidal PEG-coated magnetite nanospheres in the rat following interstitial administration.

PURPOSE: Nanoparticles can be utilised for targeting drugs to the regional lymph nodes or as diagnostic agents. The surface modification of magnetite nanospheres with poly(ethylene glycol) (PEG) has been assessed by in vitro characterisation and in vivo studies following subcutaneous administration to the rat. METHODS: Magnetite nanospheres were prepared with a grafted PEG layer using various PEG lengths from 350 to 1,000 Da. Thermogravimetric analysis was utilised to measure the adsorbed amount of PEG. Colloid stability was confirmed by measurement of the particle size and electrophoretic mobility. The kinetics of injection site drainage and lymph node retention were determined 2 hours after subcutaneous administration, for nanospheres coated with PEG lengths of 350, 550. 750, and 1,000 Da. For the 750 PEG coated nanospheres, the kinetics of distribution was determined over a 48-hour time course. RESULTS: The distribution of the nanospheres was modified and the lymph node localisation enhanced by altering the surface coverage of PEG on the magnetic surface. CONCLUSIONS: PEG-coated magnetite nanospheres with different surface characteristics can be utilised to target a diagnostic agent to regional lymph nodes.

Effects of phytoestrogens and synthetic combinatorial libraries on aromatase, estrogen biosynthesis, and metabolism.

Approximately 60% of breast cancer patients have hormone-dependent breast cancer containing estrogen receptors and requiring estrogen for tumor growth. The extent of estrogen biosynthesis and metabolism in the breast cancer tissue microenvironment influences breast-tumor development and growth, and endogenous and exogenous agents may alter the levels of hormonally active estrogens and their metabolites. Isoflavonoid phytoestrogens such as genistein exhibit numerous biochemical activities; however, their effects on estrogen biosynthesis and metabolism in breast cancer cells have not been fully examined. MCF-7 cells (hormone-dependent) and MBA-MB-231 cells (hormone-independent) were treated with genistein (100 nM) for five days and then incubated with radiolabeled estradiol (100 nM, 2.5 microCi) for 0 to 48 h. Media were extracted with ethyl acetate, and the organic residues analyzed by reverse-phase HPLC with a radioactivity flow detector. The major metabolite formed in all cases is estrone, although differences were observed between the cell lines and the various drug treatments. The formation of estrone in untreated MCF-7 cells (approximately 9.3% of radioactivity at 24 h) is relatively limited, in contrast to untreated MDA-MB-231 cells (approximately 32.0% of radioactivity at 24 h). Treatment of MCF-7 cells with 100 nM genistein increased the conversion of estradiol to estrone up to 19.5% in 24 h. The effect of genistein on estrone formation in MDA-MB-231 cells resulted in 37.7% of the radioactivity being estrone. Thus, genistein treatment of breast cancer cells resulted in increased 17-betahydroxysteroid dehydrogenase activity and elevated formation of estrone. Increased levels of oxidative 17-betahydroxysteroid dehydrogenase activity (Type II) were confirmed by Western blots. Therefore, exposure of breast cancer cells to genistein results in elevated conversion of estradiol to estrogenically weaker or inactive metabolites. The regulation of breast-tissue aromatase by exogenous agents such as drugs and environmental agents is being investigated. The benzopyranone-ring system is a molecular scaffold of considerable interest, and this scaffold is found in flavonoid natural products that have weak aromatase inhibitory activity. Medicinal chemistry efforts focus on diversifying the benzopyranone scaffold and utilizing combinatorial chemistry approaches to construct small benzopyranone libraries as potential aro- matase inhibitors. Several compounds in the initial libraries have demonstrated moderate aromatase inhibitory activity in screening assays.

Molecular pharmacology of aromatase and its regulation by endogenous and exogenous agents.

Aromatase (estrogen synthase) is the cytochrome P450 enzyme complex that converts C19 androgens to C18 estrogens. Aromatase activity has been demonstrated in breast tissue in vitro, and expression of aromatase is highest in or near breast tumor sites. Thus, local regulation of aromatase by both endogenous factors as well as exogenous medicinal agents will influence the levels of estrogen available for breast cancer growth. The prostaglandin E2 (PGE2) increases intracellular cAMP levels and stimulates estrogen biosynthesis, and our recent studies have shown a strong linear association between CYP19 expression and the sum of COX-1 and COX-2 expression in breast cancer specimens. PGE2 can bind to four receptor subtypes, EP1-EP4, which are coupled to different intracellular signaling pathways. In primary human breast stromal cell cultures, aromatase activity was significantly induced by PGE2, dexamethasone, and agonists for the EP1 and EP2 receptor subtypes. An EP1 antagonist, SC-19220, inhibited the induction of enzyme activity by PGE2 or 17-phenyltrinor-PGE2, an EP1 agonist. Sulprostone, an EP3 agonist, did not alter aromatase activity levels. Investigations are also underway on the regulation of aromatase by exogenous medicinal agents. Selective steroidal and nonsteroidal agents are effective in inhibiting breast tissue aromatase. The benzopyranone ring system is a molecular scaffold of considerable interest, and this scaffold is found in certain flavonoid natural products that have weak aromatase inhibitory activity. Our novel synthetic route for benzopyranones utilizes readily available salicylic acids and terminal alkynes as starting materials. The synthesis of flavones with diversity on the benzopyranone moiety and at the C-2 position occurs with good to excellent yields using these reaction conditions, resulting in an initial benzopyranone library of thirty compounds exhibiting enhanced and differential aromatase inhibition. Current medicinal chemistry efforts focus on diversifying the benzopyranone scaffold and utilizing combinatorial chemistry approaches to construct small benzopyranone libraries as potential aromatase inhibitors.

Properties of the initial reaction of bleomycin and several of its metal complexes with Ehrlich cells.

Characteristics of the reaction of bleomycin-A2 (BLM) and several of its metal complexes with Ehrlich cells in culture are described. Short incubation of BLM and Fe(III)-, Cu-, Zn-, and CdBLM with Ehrlich cells effectively inhibits cell proliferation. There is a sharp break at 30 min in the dependence of cytotoxicity upon time of exposure of cells to these forms of the drug. Qualitatively, the same curve can be generated by sequential additions of CoCl2 to cells during their first hour of incubation with BLM or Fe(III)BLM. The cobalt salt has less effect on CuBLM. The kinetics of initiation of the effect are directly correlated with the rapid kinetics of uptake of [3H]BLM by cells. Measurements of the initial rate of association of drug with cells as a function of extracellular BLM concentration suggest that a binding step is involved, for the rate of association approaches a maximal velocity at large concentrations of BLM. Uptake leads to both specific and nonspecific binding of tritium label; however, very little BLM gets into these cells. The internal concentration is estimated to be less than that in the external medium. BLM and its Fe(III) and copper complexes are taken up by Ehrlich cells to the same general extent after 60 min incubation; the cellular uptake of CoBLM is 25-50 times higher. Even the distributions of Fe(III)-, Cu-, and metal-free BLM within cytosol are comparable. A fraction binds to macromolecules; the rest appears unbound in low molecular weight fractions. The binding of [3H]BLM to cells cannot be reversed by incubation of labeled cells in drug-free medium or in media containing large concentrations of cold BLM.

DNA repair in skin of lupus erythematosus following in vivo exposure to ultraviolet radiation.

The in vivo repair of pyrimidine dimers induced in the DNA of skin of 9 patients diagnosed as systemic or discoid lupus erythematosus (LE) was measured. A small area of the buttock was exposed to radiation emitted from a Burdick UV-800 sunlamp. The number of pyrimidine dimers was measured by incubating the epidermal skin DNA with UV-specific endonuclease and sedimenting the DNA through alkaline sucrose gradients. The initial number of dimers induced following sunlamp exposure was 7.6 +/- 1.8 per 10(8) daltons DNA. The level of photorepair was measured by illuminating an area of the skin with greater than 450-nm radiation immediately following sunlamp exposure. We found that 56.5 +/- 9.5% of the dimers are photorepaired with 5 min of illumination. Excision repair was measured in an area of the skin covered for 2 and 24 h postirradiation. Approximately 44 and 81% of the dimers induced immediately following sunlamp exposure were removed at these respective times. These observations in LE are similar to those observed in the skin of normal individuals.

Photorepair of pyrimidine dimers in human skin in vivo.

The exposure of human skin in vivo to UV radiation emitted from a sunlamp induces the formation of pyrimidine dimers. The number of dimers, as detected by UV-endonuclease, decreases following exposure of the UV-irradiated skin to visible wavelengths of light. These results suggest that humans possess a mechanism by which pyrimidine dimers are photorepaired upon illumination of human skin in vivo with visible light.

Excision repair of UV-induced pyrimidine dimers in human skin in vivo.

The induction and loss of pyrimidine dimers in human skin in vivo was determined using UV endonuclease, alkaline sucrose sedimentations, and the fluorescent detection of nonradiolabeled DNA. The number of dimers induced following exposure of the skin to radiation emitted from a Burdick UV-800 sunlamp was quantitated by reacting the extracted DNA with Micrococcus luteus endonuclease specific for pyrimidine dimers. Exposure to 15 and 30 seconds of radiation emitted from this lamp produced the formation of 12.8 and 23.6 dimers per 10(8) daltons DNA, respectively. Approximately 50% of the dimers induced were lost 58 min after irradiation. Only a small percentage (less than 10) remained 24 hr postirradiation. These data partially characterize the process by which pyrimidine dimers are excised from human skin DNA in vivo.

Description, performance, and wavelengths of iodine stabilized lasers.

A description is given of lasers stabilized to components of the (129)I(2) spectrum in the region of the 633-nm laser lines for (3)He-(20)Ne and (3)He-(22)Ne. Relationships between operational characteristics such as power output, peak size, and peak width are shown, along with their relationships to some of the controllable parameters such as excitation level, iodine absorption, and iodine pressure. We found an iodine pressure broadening of about 13 MHz/Torr with a 2.6-MHz zero-pressure intercept. The frequency shift associated with iodine pressure is roughly 2 x 10(-9) nu/Torr to the red. Power broadening and power shifts are small, about a 10% increase in width and about 2 x 10(-11) nu variation in frequency for a fivefold to sixfold increase in power. These lasers exhibit a frequency stability for 10-sec sampling time of about 2 x 10(-12) nu and a resetability of about 1 x 10(-10) nu. The absolute vacuum wavelength for one iodine component has been measured against the (86)Kr standard-(3)He-(20)Ne:(129)I(2), kappa lambda = 632 991.2670 +/- 0.0009 pm. The wavelengths of several other iodine components have been determined by measuring the frequency difference between them and the (129)I(2), kappa component. Among these are (3)He-(20)Ne:(129)I(2), i lambda = 632 990.0742 +/- 0.0009 pm: and (3)He-(20)Ne:(127)I(2), i lambda = 632 991.3954 +/-0.0009 pm. These results were obtained using the Rowley-Hamon model for asymmetry in the krypton line and assume that the defined value for the standard is axssociated with the center of gravity of the line profile. The indicated uncertainties are statistical. No allowance has been included for imperfect realization of the krypton standard or for uncertainty in the asymmetry model.