Bi doped LaOCl and LaOF thin films grown by pulsed laser deposition.

Thin films of Bi3+ doped LaOCl and LaOF phosphors prepared via the pulsed laser deposition (PLD) technique in vacuum and different argon (Ar) pressures were compared in order to assess their luminescence properties. All peaks of the X-ray diffraction patterns of the films were consistent with the tetragonal structure of the LaOCl and LaOF, but in the case of LaOF the signal was weaker and not all peaks were present, suggesting some preferred orientation. Photoluminescence measurements revealed that the films exhibited emission around 344 nm for LaOCl:Bi and 518 nm for LaOF:Bi under excitations of 266 nm and 263 nm, respectively. The luminescence from the LaOF:Bi sample was less intense compared to the LaOCl:Bi sample prepared under the same conditions, which was also the case for the powder samples. The amount of ablated material present on the substrate was much less for LaOF:Bi compared to LaOCl:Bi, which is attributed to the greater bandgap and hence weaker absorption of the laser pulses for LaOF:Bi. Therefore phosphors based on LaOCl as the host material were found to be preferable over LaOF under the PLD conditions used in this study.

Improved Performance of Organic Thermoelectric Generators Through Interfacial Energetics.

The interfacial energetics are known to play a crucial role in organic diodes, transistors, and sensors. Designing the metal-organic interface has been a tool to optimize the performance of organic (opto)electronic devices, but this is not reported for organic thermoelectrics. In this work, it is demonstrated that the electrical power of organic thermoelectric generators (OTEGs) is also strongly dependent on the metal-organic interfacial energetics. Without changing the thermoelectric figure of merit (ZT) of polythiophene-based conducting polymers, the generated power of an OTEG can vary by three orders of magnitude simply by tuning the work function of the metal contact to reach above 1000 µW cm-2 . The effective Seebeck coefficient (Seff ) of a metal/polymer/metal single leg OTEG includes an interfacial contribution (Vinter /ΔT) in addition to the intrinsic bulk Seebeck coefficient of the polythiophenes, such that Seff  = S + Vinter /ΔT varies from 22.7 µV K-1 [9.4 µV K-1 ] with Al to 50.5 µV K-1 [26.3 µV K-1 ] with Pt for poly(3,4-ethylenedioxythiophene):p-toluenesulfonate [poly(3,4-ethylenedioxythiophene):poly(4-styrenesulfonate)]. Spectroscopic techniques are used to reveal a redox interfacial reaction affecting locally the doping level of the polymer at the vicinity of the metal-organic interface and conclude that the energetics at the metal-polymer interface provides a new strategy to enhance the performance of OTEGs.

Substrate temperature dependence of structure and optical properties of ZnTiO3:Er3+/Yb3+ thin films synthesized by pulsed laser deposition.

ZnTiO3:Er3+,Yb3+ thin film phosphors were successfully deposited by pulsed laser deposition (PLD) at different substrate temperatures. The distribution of the ions in the films was investigated and the chemical analysis showed that the doping ions were homogeneously distributed in the thin films. The optical response of the phosphors revealed that the reflectance percentages of the ZnTiO3:Er3+,Yb3+ vary with the silicon substrate temperature due to the differences in the thickness and morphological roughness of the thin films. Under 980 nm diode laser excitation, the ZnTiO3:Er3+,Yb3+ film phosphors displayed up-conversion emission from the Er3+ electronic transitions, with violet, blue, green, and red emission lines at 410, 480, 525, 545 and 660 nm from 2H9/2 â†’ 4I15/2, 4F7/2 â†’ 4I15/2, 2H11/2 â†’ 4I15/2, 4S3/2 â†’ 4I15/2 and 4F9/2 â†’ 4I15/2 transitions, respectively. The up-conversion emission was enhanced by increasing the silico (Si) substrate temperature during the deposition. Based on the photoluminescence properties and decay lifetime analysis, the energy level diagram was established and the up-conversion energy-transfer mechanism was discussed in detail.

A holistic review on the recent trends, advances, and challenges for high-precision room temperature liquefied petroleum gas sensors.

Liquefied petroleum gas (LPG), which is mainly composed of hydrocarbons, such as propane and butane, is a flammable gas that is considered a clean source of energy. Currently, the overwhelming use of LPG as fuel in vehicles, domestic settings, and industry has led to several incidents and deaths globally due to leakage. As a result, the appropriate detection of LPG is vital; thus, gas-sensing devices that can monitor this gas rapidly and accurately at room temperature, are required. This work reviews the current advances in LPG gas sensors, which operate at room temperature. The influences of the synthesis methods and parameters, doping, and use of catalysts on the sensing performance are discussed. The formation of heterostructures made from semiconducting metal oxides, polymers, and graphene-based materials, which enhance the sensor selectivity and sensitivity, is also discussed. The future trends and challenges confronted in the advancement of LPG room temperature operational gas sensors, and critical ideas concerning the future evolution of LPG gas sensors, are deliberated. Additionally, the advancements in the next-generation gas sensors, such as the wireless detection of LPG leakage, self-powered sensors driven by triboelectric/piezoelectric mechanisms, and artificial intelligent systems are also reviewed. This review further focuses on the use of smartphones to circumvent the use of costly instruments and paves the way for cost-efficient and portable monitoring of LPG. Finally, the approach of utilizing the Internet of Things (IoT) to detect/monitor the leakage of LPG has also been discussed, which will provide better alerts to the users and thus minimize the effects of leakages.

Luminescence and stability of Tb doped CaF2 nanoparticles.

Luminescence properties of CaF2:Tb3+ nanoparticles were studied in order to investigate the effect of CaF2 native defects on the photoluminescence dynamics of Tb3+ ions. Incorporation of Tb ions into the CaF2 host was confirmed by X-ray diffraction and X-ray photoelectron spectroscopy. Cross-relaxation energy transfer was observed from the photoluminescence spectra and decay curves upon excitation at 257 nm. However, the unusual long lifetime of the Tb3+ ion as well as the decreasing trend of emission lifetime of the 5D3 level suggested the involvement of traps, which were further investigated by using temperature-dependent photoluminescence measurements, thermoluminescence and lifetime measurements at different wavelengths. This work highlights the critical role that the CaF2 native defects play in the photoluminescence dynamics of Tb3+ ions incorporated in a CaF2 matrix. The sample doped with 10 mol% of Tb3+ ions was found to be stable under prolonged 254 nm ultraviolet irradiation.

Ultra-sensitive and selective p-xylene gas sensor at low operating temperature utilizing Zn doped CuO nanoplatelets: Insignificant vestiges of oxygen vacancies.

p-xylene is a harmful volatile organic compound that needs to be tested for indoor air quality detection. We report on the sensing characteristics of CuO and Zn doped CuO nanoplatelets of various concentrations that were prepared by hydrothermal synthesis, against nine different gases. These CuO and Zn based nanoplatelets were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, photoluminescence emission and vibrating sample magnetometer measurements. CuO and 0.1 at. % Zn doped CuO samples were most sensitive and selective to p-xylene gas with relatively high responses (Ra/Rg ratio) of about 42 and 53 at an operating temperature of 150 °C, respectively. These responses were about six times higher compared to the other 8 tested interfering gases. All these samples further exhibited a paramagnetic behaviour at room temperature, due to small traces of point defects, such as oxygen vacancies. Both these sensor materials did not show green luminescence at room temperature that is normally associated with oxygen vacancies. However, temperature dependent photoluminescence (PL) measurements for the 0.1 at. % Zn doped CuO showed broad visible emission, including green luminescence, which increased with temperature up to 150 °C and coincided with the gas sensing temperature. The pure CuO, however, showed a rapid quenching in PL emission with an increase in the temperature up to 150 °C. Nevertheless, both pure CuO and 0.1 at. % Zn doped CuO based sensors were highly sensitive to the p-xylene gas. The mechanism associated to the xylene superior sensing was considered in terms of point defects and surface area as active sites for adsorption of gas molecules.

Influence of Ag, Au and Pd noble metals doping on structural, optical and antimicrobial properties of zinc oxide and titanium dioxide nanomaterials.

Oxide materials (ZnO, TiO2) doped with noble metals were synthesized using the combustion technique. The results of the addition of Ag, Au, and Pd up to a concentration of 2 mol% on the structural, optical, morphological and antimicrobial properties was considered. X-ray diffraction experiments revealed that the crystal structure of the host materials remained unaltered despite doping with noble metals. From the scanning electron microscopy results, it was evident that the doped nanoparticles aggregated in clusters of different sizes in the host matrix. The plasmonic effect was also observed in the absorbance spectra of the different doped materials. The obtained materials have shown promising antimicrobial features. All ZnO materials exhibited a high antimicrobial activity, with very low minimum inhibitory concentration values, against the planktonic growth of all tested Gram-positive and Gram-negative bacterial strains. All doped materials exhibited very good anti-biofilm activity, the lowest minimal biofilm eradication concentration values being registered for ZnO doped with Au and Pd toward Escherichia coli and for ZnO doped with Ag against Candida albicans. These results indicate the potential that these materials have for antimicrobial applications in the fields of biomedicine and environmental protection.

Highly efficient infrared to visible up-conversion emission tuning from red to white in Eu/Yb co-doped NaYF4 phosphor.

Eu/Yb co-doped NaYF4 phosphors have been synthesized by the combustion method. The Eu doping was fixed and the effect of Yb doping concentration on the structural, morphological and luminescence properties has been investigated. X-ray diffraction analysis revealed that the phosphors consisted of mixed α- and ß-phases, but the ß-phase was dominant. All elements of the host and dopants, as well as adventitious C, were detected using X-ray photoelectron spectroscopy. The surface morphology showed a microrod-like structure with sharp hexagonal edges. Energy dispersive X-ray spectroscopy spectra proved the formation of the desired materials. The photoluminescence spectra illustrated the optical emission properties of Eu3+ in the red region when excited at 394 nm, while, under the same excitation, Yb3+ ions gave emission at 980 nm. The up-conversion (UC) emission of Eu/Yb co-doped NaYF4 produced a white color at the higher concentration of Yb excited by a 980 nm laser, which was made possible by green emission of Er contamination (from Yb source) and blue emission of Eu2+ ions. The lifetime of the Eu3+ UC luminescence at 615 nm was also affected by the Yb doping concentration. The temperature sensitivity associated with the Er3+ peaks at 520 and 542 nm was assessed as a function of temperature and the maximum of 0.0040 K-1 occurred at 463 K.

Imaging the atomic structure and local chemistry of platelets in natural type Ia diamond.

In the past decades, many efforts have been devoted to characterizing {001} platelet defects in type Ia diamond. It is known that N is concentrated at the defect core. However, an accurate description of the atomic structure of the defect and the role that N plays in it is still unknown. Here, by using aberration-corrected transmission electron microscopy and electron energy-loss spectroscopy we have determined the atomic arrangement within platelet defects in a natural type Ia diamond and matched it to a prevalent theoretical model. The platelet has an anisotropic atomic structure with a zigzag ordering of defect pairs along the defect line. The electron energy-loss near-edge fine structure of both carbon K- and nitrogen K-edges obtained from the platelet core is consistent with a trigonal bonding arrangement at interstitial sites. The experimental observations support an interstitial aggregate mode of formation for platelet defects in natural diamond.

Influence of Bi doping on the structure and photoluminescence of ZnO phosphor synthesized by the combustion method.

Bismuth doped ZnO (BZO) phosphors have been synthesized by the combustion method. The effect of Bi doping up to 4mol% on the structural, morphological, optical and photoluminescence (PL) properties have been investigated. X-ray diffraction analysis revealed that the BZO phosphors had the hexagonal wurtzite structure. The nanocrystallite size decreased from 75 to 38nm as the Bi concentration increased up to 3mol%, but then increased slightly for 4mol% Bi. The chemical states of the synthesized BZO phosphors were investigated using X-ray photoelectron spectroscopy and revealed the presence of both Bi3+ and Bi2+ charge states. The surface morphology showed spherical grains with some small particle agglomeration. The grain agglomeration and irregular shapes increased with increasing Bi concentration in the BZO phosphor. The absorption spectra were calculated from the reflection spectra using the Kubelka-Munk function and a blue shift in the absorption was obtained. The optical bandgap varied from 3.08 to 3.11eV for increasing Bi doping concentration. The PL spectra showed a blue emission at 410-500nm and a broad red peak at 650nm. These peaks are attributed to oxygen related defects in the ZnO host. The addition of Bi decreased the red emission and enhanced the blue emission.

Transparent conducting ZnO-CdO mixed oxide thin films grown by the sol-gel method.

Mixed oxides of zinc and cadmium with different proportions were deposited on ordinary glass substrates using the sol-gel spin coating method under optimized deposition conditions using zinc acetate dihydrate and cadmium acetate dihydrate as precursors. X-ray diffraction patterns confirmed the polycrystalline nature of the films. A combination of cubic CdO and hexagonal wurtzite ZnO phases was observed. The oxidation states of Zn, Cd and O in the deposited films were determined by X-ray photoelectron spectroscopic studies. Surface morphology was studied by scanning electron microscopy and atomic force microscopy. The compositional analysis of the thin films was studied by secondary ion mass spectroscopy. The transmittance of the thin films was measured in the range 300-800nm and the optical bandgap was calculated using Tauc's plot method. The bandgap decreased from 3.15eV to 2.15eV with increasing CdO content. The light emission properties of the ZnO:CdO thin films were studied by photoluminescence spectra recorded at room temperature. The current-voltage characteristics were also assessed and showed ohmic behaviour. The resistance decreased with increasing CdO content.

Continuous treatment with odanacatib for up to 8 years in postmenopausal women with low bone mineral density: a phase 2 study.

UNLABELLED: The efficacy and safety of weekly oral odanacatib (ODN) 50 mg for up to 8 years were assessed in postmenopausal women with low bone mineral density (BMD). Treatment with ODN for up to 8 years resulted in continued or maintained increases in BMD at multiple sites and was well tolerated. INTRODUCTION: ODN is a selective inhibitor of cathepsin K. In a 2-year phase 2b study (3/10/25/50 mg ODN once weekly [QW] or placebo) and extensions (50 mg ODN QW or placebo), ODN treatment for 5 years progressively increased BMD and decreased bone resorption markers in postmenopausal women with low BMD ( ClinicalTrials.gov NCT00112437). METHODS: In this prespecified interim analysis at year 8 of an additional 5-year extension (years 6 to 10), patients (n = 117) received open-label ODN 50 mg QW plus weekly vitamin D3 (5600 IU) and calcium supplementation as needed. Primary end points were lumbar spine BMD and safety. Patients were grouped by ODN exposure duration. RESULTS: Mean (95 % confidence interval [CI]) lumbar spine BMD changes from baseline were 4.6 % (2.4, 6.7; 3-year continuous ODN exposure), 12.9 % (8.1, 17.7; 5 years), 12.8 % (10.0, 15.7; 6 years), and 14.8 % (11.0, 18.6; 8 years). Similar patterns of results were observed for BMD of trochanter, femoral neck, and total hip versus baseline. Geometric mean changes from baseline to year 8 for bone resorption markers were approximately -50 % (uNTx/Cr) and -45 % (sCTx), respectively (all groups); bone formation markers remained near baseline levels. No osteonecrosis of the jaw, delayed fracture union, or morphea-like skin reactions were reported. CONCLUSIONS: Treatment with ODN for up to 8 years resulted in gains in BMD at multiple sites. Bone resorption markers remained reduced, with no significant change observed in bone formation markers. Treatment with ODN for up to 8 years was well tolerated.

Use of ZnO:Tb down-conversion phosphor for Ag nanoparticle plasmon absorption using a He-Cd ultraviolet laser.

Although noble metal nanoparticles (NPs) have attracted some attention for potentially enhancing the luminescence of rare earth ions for phosphor lighting applications, the absorption of energy by NPs can also be beneficial in biological and polymer applications where local heating is desired, e.g. photothermal applications. Strong interaction between incident laser light and NPs occurs only when the laser wavelength matches the NP plasmon resonance. Although lasers with different wavelengths are available and the NP plasmon resonance can be tuned by changing its size and shape or the dielectric medium (host material), in this work, we consider exciting the plasmon resonance of Ag NPs indirectly with a He-Cd UV laser using the down-conversion properties of Tb(3+) ions in ZnO. The formation of Ag NPs was confirmed by X-ray diffraction, transmission electron microscopy and UV-vis diffuse reflectance measurements. Radiative energy transfer from the Tb(3+) ions to the Ag NPs resulted in quenching of the green luminescence of ZnO:Tb and was studied by means of spectral overlap and lifetime measurements. The use of a down-converting phosphor, possibly with other rare earth ions, to indirectly couple a laser to the plasmon resonance wavelength of metal NPs is therefore successfully demonstrated and adds to the flexibility of such systems. Copyright © 2016 John Wiley & Sons, Ltd.

Optical and surface enhanced Raman scattering properties of Au nanoparticles embedded in and located on a carbonaceous matrix.

Au nanoparticles (NPs) on the surface and embedded in a matrix have been the subject of studies dealing with a variety of spectroscopic and sensing applications. Here, we report on low energy Ar ion induced evolution of the morphology of a thin Au film on a polyethylene terephthalate (PET) substrate along with thermodynamic interpretations, and corresponding unique surface plasmon resonance (SPR) and photoluminescence (PL) properties. These properties are linked to the variation of surface nanostructures and the surface enhanced Raman scattering (SERS) effect of methyl orange (MO) dye molecules adsorbed on the surface. Ion induced thermal spike and sputtering resulted in dewetting of the film with subsequent formation of spherical NPs. This was followed by embedding of the NPs in the modified PET due to the thermodynamic driving forces involved. The surface and interface morphologies were studied using atomic force microscopy and cross-sectional transmission electron microscopy. X-ray photoelectron spectroscopy was used to study the chemical changes in the system upon irradiation. The optical properties were studied by diffuse reflectance UV-Vis spectroscopy and PL using a 325 nm He-Cd laser. The red shift of the SPR absorption and the blue shift of the PL emission have been correlated with the surface morphology. The blue PL emission bands at around 3.0 eV are in good agreement with the literature with respect to the morphological changes and the blue shift is attributed to compressive strain on the embedded Au NPs. Enhancement of the SERS signals is observed and found to be correlated with the SPR response of the Au nanostructures. The SERS analyses indicate that MO molecules may be adsorbed with different orientations on these surfaces i.e. Au NPs located on the surface or embedded in the modified PET. These polymeric substrates modified by NPs can have a potential application in solid-state light emitting devices and can be applied in SERS based sensors for the detection of organic compounds.

The beta-gal interferon assay: a new, precise and sensitive method.

Cells of a human glioblastoma line were stably transfected with a glial fibrillary acidic protein (GFAP) promoter sequence/lacZ reporter gene. Following this modification, they produced Escherichia coli beta-galactosidase constitutively in amounts that could be measured through their conversion of an added fluorophore into a product readily estimated by fluorimetry. Human interferons (IFN) selectively and in a dose-dependent manner reduce the formation of beta-galactosidase in this system. We have used it as the basis for a novel assay that is sensitive (4-40 pg/ml), precise, completed in 30 h, and applicable to both type I and type II human IFNs. Statistical analysis showed interassay relative standard deviations ranging from 5% to 11%, and most individual assays revealed potencies with limits of error within 85%-115%. Neither partially trypsin-digested IFN nor the other cytokines and mitogens we tested reacted in this system, except for tumor necrosis factor-alpha (TNF-alpha). The high selectivity was further shown by the loss of response to IFN in the presence of the appropriate specific anti-IFN or anti-IFN-gamma receptor antibodies.

Effects of liver blood flow on the pharmacokinetics of tissue-type plasminogen activator (alteplase) during thrombolysis in patients with acute myocardial infarction.

BACKGROUND: The removal of recombinant tissue-type plasminogen activator (rt-PA; alteplase) by the liver is so rapid that liver blood flow becomes rate determining for its clearance. In patients with myocardial infarction changes in liver blood flow may result from impaired cardiac performance or drug treatment. OBJECTIVE: To estimate the effect of variations in liver blood flow on t-PA plasma concentrations during thrombolytic therapy. METHODS: Fifteen patients with acute myocardial infarction were investigated in an open single-center study at the coronary care unit of University Hospital Leiden. Patients received thrombolytic treatment with 100 mg rt-PA over 3 hours. Liver blood flow was estimated by indocyanine green clearance and by Doppler echocardiography. Concentrations of t-PA antigen, t-PA activity, indocyanine green, alpha 2-antiplasmin, fibrinogen, and fibrin and fibrinogen degradation products were measured. RESULTS: Indocyanine green clearance and clearance of both t-PA antigen (r = 0.78; p < 0:01) and t-PA activity (r = 0.54; p < 0.05) were significantly related. Significant associations between t-PA antigen and fibrin and fibrinogen degradation products and between t-PA antigen and alpha 2-antiplasmin were also found. CONCLUSIONS: The liver blood flow of patients with myocardial infarction is inversely correlated with plasma concentrations of t-PA. In patients with severely impaired liver blood flow and heart failure, high t-PA plasma concentrations may occur if standard doses are given. This finding could contribute to optimization of the dosage of t-PA in certain patient groups.

Effect of changes in liver blood flow on the pharmacokinetics of saruplase in patients with acute myocardial infarction.

BACKGROUND: The recombinant unglycosylated single chain urokinase-type plasminogen activator saruplase is cleared for a large part by the liver. A large interindividual variation in saruplase concentration is found in acute myocardial infarction (AMI) patients. The variable cardiac performance after an infarct may induce differences in liver blood flow that could explain the concentration diversity. This study was performed to investigate the relation between hepatic blood flow and the pharmacokinetic and pharmacodynamic properties of saruplase. METHODS AND RESULTS: Thirteen AMI patients were enrolled in this open label study. Patients received a bolus injection of 20 mg saruplase followed by a one-hour infusion of 60 mg saruplase. Concurrently 36 mg intravenous indocyanine green (ICG) was given over 1 h to measure hepatic blood flow. Blood samples were taken at regular time intervals to measure plasma levels of urokinase-type plasminogen activator (u-PA) antigen and activity, the two-chain form (tcu-PA) activity, indocyanine green, fibrinogen, fibrin and fibrin degradation products, alpha2-antiplasmin and thrombin antithrombin III complex. A correlation was seen between the clearance of ICG and both those of u-PA antigen (r = 0.62; p <0.05) and u-PA activity (r = 0.57; p <0.05). A negative correlation was seen between the area under the curve of tcu-PA activity and the areas under the effect curves of both fibrinogen and alpha2-antiplasmin (r = -0.84; p <0.01 and r = -0.65; p <0.05). CONCLUSIONS: Liver blood flow is an important determinant of the clearance of u-PA antigen and activity and reduction of flow in patients with heart failure will lead to an increase in plasma concentrations. High plasma concentrations of tcu-PA activity lead to increased systemic fibrinogenolysis. These results may be used to optimize saruplase treatment in patients with impaired cardiac function or after co-medication with drugs that affect liver blood flow.

Effects of increased liver blood flow on the kinetics and dynamics of recombinant tissue-type plasminogen activator.

OBJECTIVE: To investigate the influence of increased liver blood flow on the pharmacokinetics and pharmacodynamics of recombinant tissue-type plasminogen activator (rt-PA) and to study the changes in endogenous urokinase-type plasminogen activator (u-PA). METHODS: This open, randomized, crossover trial was carried out in a clinical research unit. Eight healthy, nonsmoking volunteers received linear infusions of 24 mg rt-PA and 92 mg indocyanine green over 160 minutes. Sixty minutes after the infusions were started, the subjects consumed a standardized meal to increase liver blood flow on one occasion and abstained from taking food on the other occasion. Plasma concentrations of indocyanine green, tissue-type plasminogen activator (t-PA) antigen, t-PA activity, total u-PA antigen, plasmin-activatable single-chain u-PA (scu-PA), active two-chain u-PA (tcu-PA), fibrinogen, total fibrin, and fibrinogen/fibrin degradation products (TDP), and alpha 2-antiplasmin were measured. RESULTS: After the consumption of the meal, the area under the curve (AUC) was 35% (95% confidence interval [CI]: 25%, 43%) lower for indocyanine green, 15% (CI: 6%, 24%) lower for t-PA antigen, and 11% (CI: 2%, 19%) lower for t-PA activity compared to the AUC after subjects abstained from food. No changes were observed in fibrinogen, TDP, or alpha 2-antiplasmin concentrations that were attributable to the intake of food. The infusion of rt-PA caused a fivefold increase in the concentration of active tcu-PA and a concomitant decrease in scu-PA concentrations by more than 50%. CONCLUSIONS: Increased liver blood flow results in an increase in t-PA clearance. The conversion of the inactive zymogen scu-PA to the active tcu-PA is increased by an infusion of rt-PA, but total u-PA antigen concentrations remain unchanged.

In vitro bioassay for human erythropoietin based on proliferative stimulation of an erythroid cell line and analysis of carbohydrate-dependent microheterogeneity.

The human erythroleukemia cell line TF-1 was employed for the determination of proliferative stimulation induced by recombinant human erythropoietin (rhEpo). Potencies of various intact and sugar-trimmed rhEpo preparations were estimated using the International Standard for Human r-DNA-derived Epo (87/684) as a reference for activity. The cellular response was measured in a multi-channel photometer using a colorimetric microassay, based on the metabolism of the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to formazan, by viable cells. The linear part of the log dose-response relationship encompassed 2.5-90 pM and activity of rhEpo preparations was measured at doses between 3 and 60 pM. The assay was designed as a parallel line test, using three or four concentrations for potency determinations, which fulfills pharmacopoeial requirements for assay validity. Inter-assay relative standard deviation varied between 4.1% and 12.6% and most assays revealed potencies with limits of error within 87-113%. In order to acquire an additional means for an efficient probing of physiologically relevant features of rhEpo, a luminiscence-dependent Western detection system, based on a combined isoelectric focusing/sodium dodecyl sulphate-polyacrylamide gel electrophoresis separation, was established. As opposed to conventional electrophoresis the two dimensional approach enabled the disclosure of minor truncations in the rhEpo-attached glycan moieties using picomolar quantities of the hormone. Moreover, the separated isoforms of rhEpo were quantified by computer-assisted densitometry and compared with the 87/684 standard. Accordingly, results obtained by the cellular response were balanced against the general pattern observed and the relative amounts of separated rhEpo isomers as determined by the quantitative Western analysis. The method described should be suitable for potency assessments of pharmaceutical formulations of rhEpo.

Tolrestat pharmacokinetic and pharmacodynamic effects on red blood cell sorbitol levels in normal volunteers and in patients with insulin-dependent diabetes.

OBJECTIVES: To examine the effect of diabetes mellitus on the pharmacokinetics of tolrestat and to investigate its effect on red blood cell sorbitol levels according to a new pharmacodynamic model for this class of drugs. METHODS: Single and multiple doses of tolrestat (200 mg/twice a day) were administered to 12 patients with insulin-dependent (type I) diabetes and 12 healthy volunteers in an open parallel trial. RESULTS: Tolrestat disposition was characterized by first-order absorption and biexponential disposition: In normal subjects the terminal disposition half-life (t1/2) was 13 +/- 3 hours (mean +/- SD) and the apparent oral clearance (CL/F) was 48 +/- 9 ml/hr/kg, similar to the values in patients with type I diabetes mellitus (t1/2 = 14 +/- 4 hours; CL/F = 55 +/- 10 ml/hr/kg). Red blood cell sorbitol concentrations, which declined because of tolrestat's inhibition of aldose reductase, were characterized by an indirect-response model including a 50% inhibition constant (IC50) for production of sorbitol by aldose reductase. The removal of sorbitol (kout) was slower in patients with diabetes. The plasma IC50 averaged 2.0 +/- 1.3 micrograms/ml in normal subjects and 2.5 +/- 1.9 micrograms/ml in patients with diabetes. IC50 values expressed in free (unbound) concentrations (fu = 0.64%), which ranged from 12 to 16 ng/ml, were similar to the in vitro IC50 for inhibition of sorbitol accumulation in human red blood cells. CONCLUSIONS: Tolrestat pharmacokinetics were similar in normal subjects and in patients with diabetes; however, the patients with diabetes had higher baseline sorbitol levels (11 versus 5 nmol/ml for normal subjects) and slower sorbitol efflux rates. The proposed biochemically realistic, dynamic model characterized well the red blood cell sorbitol response patterns after administration of single and multiple doses of tolrestat.