The effects of gum arabic oral treatment on the metabolic profile of chronic renal failure patients under regular haemodialysis in Central Sudan

This study aimed at assessing the effect of gum arabic (Acacia senegal) oral treatment on the metabolic profile of chronic renal failure (CRF) patients. A total of 36 CRF patients (under regular haemodialysis) and 10 normal subjects participated in this study. The patients were randomly allocated into three groups-group A: 12 CRF patients under low-protein diet (LPD) (<40 g day(-1)) and gum arabic (50 g day(-1)) treatment; group B: 14 CRF patients under LPD and gum arabic, iron (ferrous sulphate, 200 mg day(-1)) and folic acid (5 mg day(-1)) treatment; group C (control group): 10 CRF patients under LPD and iron and folic acid treatment and group D: 10 normal volunteers (on normal diet) under daily dose of 50 g gum arabic. Each of the above treatments was continued for three consecutive months. Blood samples were collected from each subject before treatment and twice per month "pre-dialysis" for 3 months. Biochemical parameters measured were: serum urea, serum creatinine, serum uric acid, serum calcium and serum phosphorus. By the end of the 3 months of treatment, serum urea levels significantly decreased by 31.2 and 44.18% for group A and B, respectively, compared with the baseline (0.01 < p < 0.001) and control group (p < 0.05). Serum creatinine levels significantly decreased in the groups of gum users (A, B and D) by 9.94, 12.65 and 11.7%, respectively, compared with the control group (p < 0.001). There was a significant decrease (p < 0.05) in serum uric acid levels by 14 and 19.9% for group A and B, respectively, compared with the baseline. Serum calcium levels increased by 12.64, 15.75 and 8.75% for group A, B and D, respectively, and these increases were significantly different (0.05 < p < 0.001) from baseline and control group for groups A and B. Serum phosphorus levels significantly decreased by 22.54% for group A, 17.69% for group B and 7.71% for group D, compared with the baseline (0.05 < p < 0.001). From this study, we conclude that oral administration of gum arabic could conceivably alleviate adverse effects of CRF.     

Temperature‐controlled ultrasound‐ and vortex‐assisted liquid–liquid microextraction combined with GC for the determination of the concentrations of organophosphorus pesticides in beverage samples

The aim of this work was to develop temperature‐controlled ultrasound‐ and vortex‐assisted liquid–liquid microextraction as a fast and efficient approach for the extraction of nine organophosphorus pesticides in beverage samples followed by GC with flame photometric detection analysis. The combination of ultrasonication and vortexing were used to assist the microextraction, and the use of a dispersion solvent was avoided. Several variables that could potentially affect the extraction efficiency, namely, the type and volume of extraction solvent, sequence, and time of ultrasonication and vortexing, ultrasonication bath temperature and ionic strength were optimized. Under optimum conditions, the calibration graphs were linear over the range of 0.5–200 μg/L. The LOD (S/N = 3) was between 0.01 and 0.05. The optimized method exhibited a good precision level with RSD values between 4.5 and 9.8%. The enrichment factors for the nine organophosphorus pesticides were between 224 and 339. Four beverage samples were successfully analyzed using the proposed method.     

Vortex‐ and CO2‐gas‐assisted liquid–liquid microextraction with salt addition for the high‐performance liquid chromatographic determination of furanic compounds in concentrated juices and dried fruits

A novel microextraction method based on vortex‐ and CO₂‐assisted liquid–liquid microextraction with salt addition for the isolation of furanic compounds (5‐hydroxymethyl‐2‐furaldehyde, 5‐methyl‐2‐furaldehyde, 2‐furaldehyde, 3‐furaldehyde, 2‐furoic and 3‐furoic acids) was developed. Purging the sample with CO₂ was applied after vortexing to enhance the phase separation and mass transfer of the analytes. The optimum extraction conditions were: extraction solvent (volume), propyl acetate (125 μL); sample pH, 2.4; vortexing time, 45 s; salt concentration, 25% w/v and purging time, 5 min. The analytes were separated using an ODS Hypersil C₁₈ column (250×4.6 mm i.d, 5 μm) under gradient flow. The proposed method showed good linearities (r² >0.999), low detection limits (0.08–1.9 μg/L) and good recoveries (80.7–122%). The validated method was successfully applied for the determination of the furanic compounds in concentrated juice (mango, date, orange, pomegranate, roselle, mangosteen and soursop) and dried fruit (prune, date and apricot paste) samples.     

Hydrolysis of gadolinium(III) in light and heavy water

The hydrolysis constants of Gd(3+) and the solubility products of Gd(OH)(3) and Gd(OD)(3) in nitrate solutions at 25 and 70 degrees in H(2)O and D(2)O have been determined because of their importance in nuclear technology. The constants are defined (charges omitted for clarity) as *K(11) = a(GdOH)a(H)/a(Gd), *K(21) = a(Gd(OH)(2))a(2)(H)/a(Gd), *K(SO) = a(Gd)/a(3)(H). The values for the H(2)O system were p*K(11) = 7.87 +/- 0.02, p*K(21) = 15.I6 +/- 0.09, p*K(SO) = -19.32 +/- 0.03 at 25 degrees and p*K(11) = 7.55 +/- 0.03, p*K(21) = 13.04 +/- 0.03, p*K(SO) = -16.16 +/- 0.04 at 70 degrees . For the D(2)O system they were p*K(D)(11) = 8.17 +/- 0.01, p*K(D)(21) = 16.00 +/- 0.09, p*K(D)(SO) = -21.18 +/- 0.04 at 25 degrees and p*K(D)(11) = 7.84 +/- 0.02, p*K(D)(21) = 13.95 +/- 0.02, p*K(D)(SO) = -17.34 +/- 0.04 at 70 degrees . The mean enthalpy changes of the reactions were also calculated.     

Estrogen-induced cholestasis results in a dramatic increase of b-series gangliosides in the rat liver

Hepatic ganglioside composition was investigated in normal and cholestatic Wistar rats. Cholestasis was induced by 17alpha-ethinylestradiol (EE; 5 mg/kg body weight s.c. for 18 days). As compared with controls, the EE administration resulted in severe cholestasis, as indicated by biochemical as well as morphological signs. Gangliosides isolated from the liver tissue were separated by TLC, with resorcinol-HCl detection and densitometric evaluation. As compared with controls, the total hepatic lipid sialic acid content in cholestatic rats was increased almost 2-fold (44.3 +/- 15.2 vs 79.1 +/- 9.0 nmol/g wet weight of liver tissue, p < 0.01). This increase was primarily due to the increase of ganglioside GD1a (3.6 +/- 1.0 vs 11.8 +/- 3.0 nmol/g wet weight of liver tissue, p = 0.001), as well as to the enormous up-regulation of b-series gangliosides GD3 (0.08 +/- 0.03 vs 2.0 +/- 1.2 nmol/g wet weight of liver tissue, p = 0.002), GD1b (0.1 +/- 0.06 vs 5.4 +/- 1.6 nmol/g wet weight of liver tissue, p = 0.002) and GT1b (0.06 +/- 0.03 vs 6.4 +/- 2.6 nmol/g wet weight of liver tissue, p = 0.002). As the majority of gangliosides are concentrated in cell membranes, our findings suggest that dramatic increase of b-series gangliosides might contribute to the protection of hepatocytes against the deleterious effects of cholestasis.     

Measurement of pulmonary surfactant disaturated-phosphatidylcholine synthesis in human infants using deuterium incorporation from body water

The aim of the study was to determine surfactant palmitate disaturated-phosphatidylcholine (DSPC-PA) synthesis in vivo in humans by the incorporation of deuterium from total body water into DSPC-PA under steady state condition. We studied three newborns and one infant (body weight (BW) 4.6 +/- 2.9 kg, gestational age 37.5 +/- 2 weeks, age 9 +/- 9 days) and four preterm newborns (BW 1.3 +/- 0.6 kg, gestational age 30.3 +/- 2.5 weeks, postnatal age 8.8 +/- 9.2 h). All infants were mechanically ventilated during the study and the four preterm infants received exogenous surfactant at the start of the study.We administered 0.44 g (2)H(2)O/kg BW as a bolus intravenously, followed by 0.0125 g (2)H(2)O/kg BW every 6 h to maintain deuterium enrichment at plateau over 2 days. Urine samples and tracheal aspirates (TA) were obtained prior to dosing and every 6 h thereafter. Isotopic enrichment curves of DSPC-PA from sequential TA and urine deuterium enrichments were analyzed by Gas Chromatography-Isotope Ratio-Mass Spectrometry (GC-IRMS) and normalized for Vienna Standard Mean Ocean Water. Enrichment data were used to measure DSPC-PA fractional synthesis rate (FSR) from the linear portion of the DSPC-PA enrichment rise over time, relative to plateau enrichment of urine deuterium. Secretion time (ST) was defined as the time lag between the start of the study and the appearance of DSPC-PA deuterium enrichment in TA. Data were given as mean +/- SD.All study infants reached deuterium-steady state in urine. DSPC-PA FSR was 6.5 +/- 2.8%/day (range 2.6-10.2). FSR for infants who did not receive exogenous surfactant was 5.7 +/- 3.5%/day (range 2.6-9.9%/day) and 7.3 +/- 2.1%/day (range 5.1-10.2%/day) in the preterms, whereas DSPC-PA ST was 10 +/- 10 h and 31 +/- 10 h respectively.Surfactant DSPC-PA synthesis can be measured in humans by the incorporation of deuterium from body water. This study is a simpler and less invasive method compared to previously published methods on surfactant kinetics by means of stable isotopes.     

Sequential anaerobic/aerobic treatment of dye-containing wastewaters: colour and COD removals, and ecotoxicity tests

Colour and COD removals of the azo dyes Congo Red (CR) and Reactive Black 5 (RB5) were individually evaluated in a sequential anaerobic/aerobic treatment system. Additionally, dye toxicity was assessed by using acute ecotoxicity tests with Daphnia magna as the indicator-organism. The anaerobic reactor was operated at approximately 27 °C and with hydraulic retention times of 12 and 24 h. The aerobic reactor was operated in batch mode with a total cycle of 24 h. During anaerobic step, high colour removals were obtained, 96.3% for CR (400 mg/L) and 75% for RB5 (200 mg/L). During the aerobic phase, COD effluent was considerably reduced, with an average removal efficiency of 52% for CR and 85% for RB5, which resulted in an overall COD removal of 88% for both dyes. Ecotoxicity tests with CR revealed that the anaerobic effluent presented a higher toxicity compared with the influent, and an aerobic post-treatment was not efficient in reducing toxicity. However, the results with RB5 showed that both anaerobic and aerobic steps could decrease dye toxicity, especially the aerobic phase, which removed completely the toxicity in D. magna. Therefore, the anaerobic/aerobic treatment is not always effective in detoxifying dye-containing wastewaters, sometimes even increasing dye toxicity.     

Guided three-dimensional growth of functional cardiomyocytes on polyethylene glycol nanostructures

We introduce well-defined nanopillar arrays of a poly(ethylene glycol) (PEG) hydrogel as a cell culture platform to guide a 3D construct of primary rat cardiomyocytes in vitro for potential tissue engineering applications. Ultraviolet (UV)-assisted capillary lithography was used to fabricate highly uniform approximately 150 nm PEG pillars with approximately 400 nm height. It was found that cell adhesion was significantly enhanced on PEG nanopillars (132 +/- 29 cells/mm2) compared to that on the bare PEG control (39 +/- 17 cells/mm2) (p < 0.05) but substantially reduced compared to that on the glass control (502 +/- 45 cells/mm2) (p < 0.01). Furthermore, in colonizing cardiomyocytes, the nanopillars stimulated self-assembled aggregates among the contacting cells with 3D growth, which is a unique feature for nanopatterned PEG hydrogels as a cell culture substrate. The 3D-grown cardiomyocytes retained their conductive and contractile properties, as evidenced by the observation of beating cardiomyocytes with robust action potential generation.     

Stability and bioequivalence studies of two marketed formulations of coenzyme Q10 in beagle dogs.

Coenzyme Q10 (CoQ10), a highly lipophilic compound present in the inner mitochondrial membrane, is essential for production of cellular energy in the form of ATP. CoQ10 is used as an antioxidant and also in the treatment of various cardiovascular disorders. The relative bioavailabilities of powder filled capsule (I) and oil-based formulation (II) of CoQ10 were compared in beagle dogs in an open, randomized, multiple dose, cross-over design. Poor and slow absorption characteristics were observed for both the formulations. The AUC, Cmax, and Tmax for formulation I and II are comparable (p < 0.05) where the values for formulation I are 22.84 +/- 6.3 micrograms ml-1 h, 0.51 +/- 0.11 microgram/ml, and 6.1 +/- 2.0 h whereas the values for formulation II are 24.32 +/- 5.6 micrograms ml-1 h, 0.55 +/- 0.16 microgram/ml, and 6.6 +/- 2.3 h, respectively. Stability of CoQ10 at various temperature and humidity conditions and its photostability were studied. Various antioxidants were evaluated to determine the type and amount of antioxidant(s) required to improve the stability of CoQ10. Large extent of degradation was observed at 45 degrees C and 55 degrees C. The effect of humidity conditions on degradation was insignificant. Among the various antioxidants studied, mixture of ascorbic acid (5%) and EDTA (0.1%) offered better protection than phenolic antioxidants such as butylated hydroxy anisole (BHA), butylated hydroxy toluene (BHT), or propyl gallate (PG). Further, increasing concentrations of phenolic antioxidants (from 0.1 to 0.3%) accelerated the degradation.     

Kinetics of heterogeneous reactions of HO2 radical at ambient concentration levels with (NH4)2SO4 and NaCl aerosol particles

The HO2 uptake coefficient (gamma) for inorganic submicrometer wet and dry aerosol particles ((NH4)2SO4 and NaCl) under ambient conditions (760 Torr and 296 +/- 2 K) was measured using an aerosol flow tube (AFT) coupled with a chemical conversion/laser-induced fluorescence (CC/LIF) technique. The CC/LIF technique enabled experiments to be performed at almost the same HO2 radical concentration as that in the atmosphere. HO2 radicals were injected into the AFT through a vertically movable Pyrex tube. Injector position-dependent profiles of LIF intensity were measured as a function of aerosol concentration. Measured gamma values for dry aerosols of (NH4)2SO4 were 0.04 +/- 0.02 and 0.05 +/- 0.02 at 20% and 45% relative humidity (RH), respectively, while those of NaCl were <0.01 and 0.02 +/- 0.01 at 20% and 53% RH, respectively. For wet (NH4)2SO4 aerosols, measured gamma values were 0.11 +/- 0.03, 0.15 +/- 0.03, 0.17 +/- 0.04, and 0.19 +/- 0.04, at 45%, 55%, 65%, and 75% RH, respectively, whereas for wet NaCl aerosols the values were 0.11 +/- 0.03, 0.09 +/- 0.02, and 0.10 +/- 0.02 for 53%, 63%, and 75% RH, respectively. Wet (NH4)2SO4 and NaCl aerosols doped with CuSO4 showed gamma values of 0.53 +/- 0.12 and 0.65 +/- 0.17, respectively. These results suggest that compositions, RH, and phase for aerosol particles are significant to HO2 uptake. Potential HO2 loss processes and their atmospheric contributions are discussed.     

Magnetization of a Cu(II)-1,3,5-benzenetricarboxylate metal-organic framework for efficient solid-phase extraction of Congo Red

We describe a facile method for the preparation of magnetic microparticles consisting of a metal-organic framework (MOF) of Cu(II)-BTC (BTC = 1,3,5-benzenetricarboxylate) for rapid magnetic solid-phase extraction (MSPE) of the dye Congo Red (CR) from aqueous solution. Magnetization of the MOF and solid-phase extraction of CR was simultaneously accomplished by mixing, under ultrasonication, the MOF and silica-coated magnetite microparticles in the solution to be extracted. Under optimized conditions, the magnetic MOF hybrid exhibits a fast adsorption rate and a high removal efficiency (>97 %) toward CR even over 15-fold reuse, which also features high adsorption capacity of 97.7 and 92.5 % for cationinc Methylene Blue (MB) and Crystal Violet (CV), respectively. The desorption of CR from the magnetic MOF was realized by washing it with ethanol and water. The material is considered to be a promising new adsorbent for use in wastewater treatment and in analytical preconcentration.

We report on a method for magnetization of a Cu-BTC metal-organic framework for use in rapid magnetic solid phase extraction of Congo Red from aqueous solutions. The material maintains its high removal efficiency (of >97 %) over 15 recyclable experiments.

     

Development and characterization of an all-solid-state potentiometric biosensor array microfluidic device for multiple ion analysis

A microfluidic device with an all-solid-state potentiometric biosensor array was developed using microfabrication technology. The sensor array included a pH indicator, and potassium and calcium ion-selective microelectrodes. The pH indicator was an iridium oxide thin film modified platinum microelectrode and the iridium oxide was deposited by an electrochemical method. The potassium and calcium ion-selective microelectrodes were platinum coated with silicon rubber based ion-selective membranes with respectively potassium (valinomycin) and calcium (ETH 1001) ionophores. The detection system was integrated with a micro-pneumatic pump which can continuously drive fluids into the microchannel through sensors at flow rates ranging from 52.4 microl min(-1) to 7.67 microl min(-1). The sensor array microfluidic device showed near-Nernstian responses with slopes of 62.62 mV +/- 2.5 mV pH(-1), 53.76 mV +/- 3 mV -log[K+](-1) and 25.77 mV +/- 2 mV -log[Ca2+](-1) at 25 degrees C +/- 5 degrees C, and a linear response within the pH range of 2-10, with potassium and calcium concentrations between 0.1 M and 10(-6) M. In this study the device provided a convenient way to measure the concentration of hydrogen, potassium and calcium ions, which are important physiological parameters.     

Integrated microfluidic devices with enhanced separation performance: application to phosphoproteome analyses of differentiated cell model systems

This work reports on the application of a microfluidic device integrating nanoscale LC to nanoelectrospray MS (nano-LC-chip-MS) for the analysis of complex protein digests. Peak profile analyses of more than 700 peptide ions, reproducibly detected across replicate nano-LC-chip-MS runs (n = 5), indicated that the system provided RSD values of 0.24% on retention time, +/- 30 ppm on m/z measurement and +/- 30% variation on intensity over three orders of magnitude. RP adsorbant media with different alkyl chains and particle size packed in both trapping and separation channels were investigated to improve the chromatographic performance of this system. A two-fold improvement in chromatographic peak capacity was achieved using microfluidic devices comprising a 5 mircrom C3 trap with 2.5 microm C18 trap separation channel compared to the traditional 5 microm C18 stationary phase. Enhanced sample selectivity for the identification of phosphopeptides was obtained by combining immobilized metal affinity media prior to peptide separation on the RP microfluidic device. This system was evaluated in the context of differential phosphoproteome analyses to identify changes in signaling events and protein expression of human monocytes following the administration of phorbol ester.     

Soft lithographic patterning of supported lipid bilayers onto a surface and inside microfluidic channels

We present simple soft lithographic methods for patterning supported lipid bilayer (SLB) membranes onto a surface and inside microfluidic channels. Micropatterns of polyethylene glycol (PEG)-based polymers were fabricated on glass substrates by microcontact printing or capillary moulding. The patterned PEG surfaces have shown 97 +/- 0.5% reduction in lipid adsorption onto two dimensional surfaces and 95 +/- 1.2% reduction inside microfluidic channels in comparison to glass control. Atomic force microscopy measurements indicated that the deposition of lipid vesicles led to the formation of SLB membranes by vesicle fusion due to hydrophilic interactions with the exposed substrate. Furthermore, the functionality of the patterned SLBs was tested by measuring the binding interactions between biotin (ligand)-labeled lipid bilayer and streptavidin (receptor). SLB arrays were fabricated with spatial resolution down to approximately 500 nm on flat substrate and approximately 1 microm inside microfluidic channels, respectively.     

Kinetic studies of the reactions of O2(b 1sigma(g)+) with several atmospheric molecules

Thermal rate coefficients for the removal (reaction + quenching) of O2(1sigma(g)+) by collision with several atmospheric molecules were determined to be as follows: O3, k3(210-370 K) = (3.63 +/- 0.86) x 10(-11) exp((-115 +/- 66)/T); H2O, k4(250-370 K) = (4.52 +/- 2.14) x 10(-12) exp((89 +/- 210)/T); N2, k5(210-370 K) = (2.03 +/- 0.30) x 10(-15) exp((37 +/- 40)/T); CO2, k6(298 K) = (3.39 +/- 0.36) x 10(-13); CH4, k7(298 K) = (1.08 +/- 0.11) x 10(-13); CO, k8(298 K) = (3.74 +/- 0.87) x 10(-15); all units in cm3 molecule(-1) s(-1). O2(1sigma(g)+) was produced by directly exciting ground-state O2(3sigma(g)-) with a 762 nm pulsed dye laser. The reaction of O2(1sigma(g)+) with O3 was used to produce O(3P), and temporal profiles of O(3P) were measured using VUV atomic resonance fluorescence in the presence of the reactant to determine the rate coefficients for removal of O2(1sigma(g)+). Our results are compared with previous values, where available, and the overall trend in the O2(1sigma(g)+) removal rate coefficients and the atmospheric implications of these rate coefficients are discussed. Additionally, an upper limit for the branching ratio of O2(1sigma(g)+) + CO to give O(3P) + CO2 was determined to be < or = 0.2% and this reaction channel is shown to be of negligible importance in the atmosphere.     

Myocardial injury occurs earlier than myocardial inflammation in acute experimental viral myocarditis

Endomyocardial biopsy often fails to show myocardial inflammation for patients with clinically suspected myocarditis. The serum isoforms of troponin T (cTnT) level is a very sensitive marker of myocardial injury and it is elevated even in the absence of myocardial inflammation. We investigated the correlations for myocardial injury, virus titers and inflammation in acute viral infection. Using the murine coxsackievirus group B3 (CVB3) myocarditis model, the histopathologic findings and virus titers in mouse hearts were compared with the serum cTnT levels measured by ELISA at various time points. Viable virus titers in the hearts peaked at 3 days after infection (8.22 +/- 0.13 log10 PFU/100 mg of heart); they decreased at day 7 and no viable virus was detected from day 14. Myocardial inflammation was minimal at day 3, peaked at day 7 and markedly decreased at day 14. The individual serum TnT levels were significantly increased at day 3 (7.37 +/- 1.46 ng/ml), persisted to day 7 (0.73 +/- 0.08 ng/ml), and normalized at day 14. Serum cTnT levels were correlatable with virus titers in the heart (r = 0.744, P <0.01), but the serum cTnT levels were not correlated with the degrees of inflammation. Using the less myocarditic strain of CVB3, similar relationships were observed between the changes for the serum cTnT levels and the heart virus titers. During the course of viral infection, myocardial injury precedes the pathologic evidence of inflammation, and the elevated cTnT levels provide evidence of myocardial injury even in the absence of any histologic findings of myocarditis.     

Determination of arginine and asymmetric dimethylarginine (ADMA) in human plasma by liquid chromatography/mass spectrometry with the isotope dilution technique

Arginine (ARG) is a substrate for endogenous nitric oxide (NO) production whereas its metabolite, asymmetric dimethylarginine (ADMA), acts as an inhibitor. Sufficient NO production is essential for cardiovascular key functions, thus elevated concentration levels of ADMA are related to a range of cardiovascular diseases. Owing to the lack of reliable methods for the measurement of ARG and ADMA in human plasma, concentration values determined with these methods can differ considerably. We present here a simple and very robust liquid chromatographic/mass spectrometric method for the determination of ARG and ADMA utilizing isotope-labeled internal standards. Sample preparation requires only protein precipitation; the analytes were derivatized with o-phthalaldehyde-mercaptoethanol and separated on a reversed-phase C(18) column with gradient elution. The analytes were detected with an electrospray ionization ion trap instrument working in the full-scan single mass spectrometry mode. Concentration values obtained with this method for healthy controls were ARG = 63.9 +/- 23.9 microM and ADMA = 0.355 +/- 0.066 microM, with a normal range for ADMA from 0.225 to 0.485 microM. The corresponding values for end-stage chronic renal failure patients are ARG = 48.1 +/- 18.5 microM, p < 0.01 and ADMA = 0.673 +/- 0.134 M, p < 0.001.     

A microfluidic bioreactor for increased active retrovirus output

Retroviruses are one of the most commonly used vectors in ongoing gene therapy clinical trials. To evaluate and advance virus production on the microscale platform, we have created a novel microfluidic bioreactor for continuous retrovirus production. We investigated the growth kinetics of a retroviral packaging cell line in microfluidic bioreactors for several compartment sizes, and packaging cells perfused in the microdevices showed similar growth kinetics to those cultured in conventional static conditions. To evaluate the efficiency of retrovirus production, virus titers from the microdevices were compared to those obtained from static tissue culture. When retrovirus production and collection were maintained at 37 degrees C, virus production levels were comparable for the microdevices and static tissue culture conditions. However, immediate cold storage downstream of the packaging cells in the microdevices resulted in 1.4- to 3.7-fold greater active virus production levels with the microdevices compared to the conventional static conditions over a 5 day period. Lastly, the use of microfluidics for virus production provides a continuous supply of virus supernatant for immediate infection of target cells or for preservation and storage. Such devices will be valuable for the optimization of production and evaluation of retroviruses and other viral vectors for gene therapy applications.     

High performance liquid chromatographic determination of methylxanthines in canine serum, gastric and pancreatic juices

A convenient high performance liquid chromatographic method for the determination of methylxanthines in biological samples is described. Separation was achieved by reversed phase chromatography using a mobile phase consisting of tetrahydrofuran + methanol + 0.01M potassium dihydrogen phosphate, pH 3.5 (1:20:79, v/v/v), on a 7 microns C18 column and a C18 Lichrosorb precolumn at a flow rate of 0.8 mL/min. Levels varying from 0.25-16 mg/L could be detected by UV at 280 nm. In this range, standard curves were established for 4 methylxanthines: theobromine, paraxanthine, theophylline and caffeine in 4 media: mobile phase, serum, gastric and pancreatic juices, and were found to be linear (r greater than or equal to 0.9975). Overall characteristics of the method were determined as: percent recovery (89.54%), accuracy (greater than or equal to 99.4%) and reproducibility (greater than or equal to 95%). Retention times ranged from 4.21 +/- 0.01 (1-methyluric acid) to 10.8 +/- 0.03 min (caffeine). Animal experiments (5 and 10 mg/kg boluses) were used to determine caffeine half life in dog's blood (310 +/- 46 and 453 +/- 59 min, respectively) and its secretion into pentagastrin stimulated gastric juice (mean concentrations 2.51 and 6.04 mg/L; mean outputs 351 and 1206 micrograms/2.25 h; both statistically different at p less than 0.001 level).     

High-volume production of single and compound emulsions in a microfluidic parallelization arrangement coupled with coaxial annular world-to-chip interfaces

This study describes a microfluidic platform with coaxial annular world-to-chip interfaces for high-throughput production of single and compound emulsion droplets, having controlled sizes and internal compositions. The production module consists of two distinct elements: a planar square chip on which many copies of a microfluidic droplet generator (MFDG) are arranged circularly, and a cubic supporting module with coaxial annular channels for supplying fluids evenly to the inlets of the mounted chip, assembled from blocks with cylinders and holes. Three-dimensional flow was simulated to evaluate the distribution of flow velocity in the coaxial multiple annular channels. By coupling a 1.5 cm × 1.5 cm microfluidic chip with parallelized 144 MFDGs and a supporting module with two annular channels, for example, we could produce simple oil-in-water (O/W) emulsion droplets having a mean diameter of 90.7 μm and a coefficient of variation (CV) of 2.2% at a throughput of 180.0 mL h(-1). Furthermore, we successfully demonstrated high-throughput production of Janus droplets, double emulsions and triple emulsions, by coupling 1.5 cm × 1.5 cm - 4.5 cm × 4.5 cm microfluidic chips with parallelized 32-128 MFDGs of various geometries and supporting modules with 3-4 annular channels.