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Pathological evaluation, clinical chemistry and plasma cholecystokinin in neonatal and young miniature swine fed soy trypsin inhibitor from 1 to 39 weeks of age.

The potential toxicity of dietary soy trypsin inhibitor (TI) was evaluated in neonatal miniature swine. From 1 to 6 weeks of age, two groups of male piglets were artificially reared in an Autosow and automatically fed either TI or control liquid diet. From 6 to 39 weeks of age, these two groups were fed either TI or control chow diet. A third group, sow control (SC), suckled from birth to 6 weeks of age, were also weaned to control chow from 6 to 39 weeks of age. Clinical chemistry and plasma cholecystokinin (CCK) determined at 6, 18, 30 and 39 weeks of age, and serum amylase activity with gross and histopathological analyses of major organs at 6 and 39 weeks of age are reported. TI had no effect on plasma CCK, serum amylase activity, or numerous clinical chemistry values. TI-fed piglets had a larger relative liver weight at 6 weeks of age. Relative pancreas weight decreased with age but was not affected by TI. Gross and histopathological analyses of major organs, except the spleen, were within normal limits. Increased incidence of extramedullary hematopoiesis was noted in the spleen of the TI group at 6 but not at 39 weeks of age. There was no consistent pattern in immunohistochemical foci for secretin, gastrin releasing polypeptide or CCK, and no change in DNA, RNA, mitotic index or nuclear density of pancreatic cells. At 6 weeks of age, TI increased pancreatic protein and amylase activity but not trypsin or chymotrypsin activity. None of the effects suggested that this dose of TI was toxic to either the neonatal or sexually mature miniature male swine.

Administration, Oral↗

Initial resolution measurements of miniaturized electrostatic lenses for LVSEM.

Theoretical and experimental investigations on miniaturized electrostatic lenses for high-resolution low-voltage scanning electron microscopical applications are presented. The electron optical column consists of a Schottky emitter including the extraction anode and a miniaturized three electrode lens consisting of conventional (electron microscopical) platinum apertures. The lens has been optimized with respect to resolution and a value of about 5 nm at a working distance of 1 mm and a beam energy of 1 keV is predicted by simulation. Details on the resolution measurements are presented. An initial experimental value amounts to 31 nm. Specific problems encountered during building and alignment of the lens and measuring the resolution are discussed as well.

Journal Article↗

Deep venous thrombosis prophylaxis in trauma: improved compliance with a novel miniaturized pneumatic compression device.

OBJECTIVE: Intermittent pneumatic compression (IPC) devices prevent lower-extremity deep venous thrombosis (LEDVT) when used properly, but compliance remains an issue. Devices are frequently discontinued when patients are out of bed, and they are rarely used in emergency departments. Trauma patients are at high risk for LEDVT; however, IPCs are underused in this population because of compliance limitations. The hypothesis of this study was that a new miniaturized, portable, battery-powered pneumatic compression device improves compliance in trauma patients over that provided by a standard device. METHODS: This was a prospective trial in which trauma patients (mean age, 46 years; revised trauma score, 11.7) were randomized to DVT prophylaxis with a standard calf-length sequential IPC device (SCD group) or a miniaturized sequential device (continuous enhanced-circulation therapy [CECT] group). The CECT device can be battery-operated for up to 6 hours and worn during ambulation. Timers attached to the devices, which recorded the time each device was applied to the legs and functioning, were used to quantify compliance. For each subject in each location during hospitalization, compliance rates were determined by dividing the number of minutes the device was functioning by the total minutes in that location. Compliance rates for all subjects were averaged in each location: emergency department, operating room, intensive care unit, and nursing ward. RESULTS: Total compliance rate in the CECT group was significantly higher than in the SCD group (77.7% vs. 58.9%, P =.004). Compliance in the emergency department and nursing ward were also significantly greater with the CECT device (P =.002 and P =.008 respectively). CONCLUSIONS: Previous studies have demonstrated that reduced compliance with IPC devices results in a higher incidence of LEDVT. Given its ability to improve compliance, the CECT may provide superior DVT prevention compared with that provided by standard devices.

Aged↗

Miniaturized real-time monitoring system for L-lactate and glucose using microfabricated multi-enzyme sensors.

A miniaturized on-line monitoring system for the detection of L-lactate and glucose is presented. The system is based on a microfabricated multi-enzyme silicon sensor chip with flow channels integrated on the chip. The sensors were fabricated in containment technology. They were characterized in test solutions. The cross-talking behaviour was investigated and was found to be practically negligible. The linear measurement ranges of both glucose and lactate sensors were large enough for most practical applications. As a result of the miniaturization the analyte consumption could be reduced to a few nmol min(-1). The system was equipped with a microdialysis probe whose recovery was 45% for lactate and 37% for glucose in test solutions using a flow rate of 3 microl min(-1). Lower flow rates of 0.5 microl min(-1) resulted in recoveries of over 90%. The long-term stability of the system was acceptable. Initial measurements have also been performed in vitro using human blood serum.

Biosensing Techniques↗

Miniaturized implantable rotary blood pump in atrial-aortic position supports and unloads the failing heart.

The purpose of this study was to test the pump performance of a miniaturized, newly developed, implantable rotary blood pump (the diagonal pump). In a series of six sheep, the pump was interposed in an atrial-aortic connection. Heart failure was induced by serial injection of glass beads into the left coronary artery. The assistance of the non-failing heart (with pump output up to 4.5 liter/min) did not change the total cardiac output or the blood pressure of the animal. The heart was significantly unloaded as demonstrated by a drop in first derivative of the left ventricular pressure (dP/dt) max (from 1645 to 1113 mmHg/s; P = 0.0003). Because of the specific interaction between heart function and rotary blood pump performance, the pump provided considerably more flow in the failing than in the non-failing heart. Cardiac output and perfusion pressures were restored to pre-assist values in the failing heart model. The heart-rotary blood pump interaction is an ideal substrate for long-term assistance. With this miniaturized rotary blood pump, hemodynamics in a severely failing heart can be restored. Atrial cannulation, which leaves the left ventricle untouched, provides similar results as apical cannulation in the failing heart.

Animals↗

Sub-miniature ExB sector-field mass spectrometer.

A novel sub-miniature double-focusing sector-field mass spectrometer has been fabricated at the University of Minnesota using a combination of conventional machining methods and thin film patterning techniques typically used in the sensor technology industry. Its design is based on the mass separation capabilities of a 90 degrees cylindrical crossed electric and magnetic sector-field analyzer with a 2-cm radius, which under proper conditions is able to effectively cancel the angular and chromatic dispersion of the ion beam, thus improving the resolving power of the instrument. Simulations using finite element analysis and computer modeling were employed to verify and optimize the performance of the proposed instrument before and during its fabrication. The prototype was able to attain a resolving power of 106 full-width at half-maximum (FWHM), a detection limit close to 10 parts per million, a dynamic range of 5 orders of magnitude and a mass range up to 103 Da. Its overall size, including the magnet assembly, is 3.5 cm wide, 6 cm long and 7.5 cm tall, it weighs 0.8 kg, and its power consumption was measured to be 2.5 W. The performance of the instrument was found to be comparable to that of commercial residual gas analyzers, at a fraction of the cost. All these characteristics make this miniature mass spectrometer suitable for portable and low-cost analytical instrumentation.

Journal Article↗

Challenges and solutions to ultra-high-throughput screening assay miniaturization: submicroliter fluid handling.

The miniaturization of HTS assays is an important objective in the pharmaceutical industry. The ability to perform primary screening assays in high-density micro-well plates at volumes of 1-2 ml will accelerate the early stages of drug discovery and reduce costs. Ultra-HTS (uHTS) assays require an accurate and reliable means of fluid handling in the submicroliter volume range. This relates to the design of instrumentation for dispensing fluids, as well as assay plates. Fluid handling has been a major obstacle to the full implementation of miniaturized assays. This report focusses on current approaches to submicroliter fluid handling in high-density multi-well plates.

Journal Article↗

High-throughput miniature cylindrical ion trap array mass spectrometer.

A fully multiplexed cylindrical ion trap (CIT) array mass spectrometer with four parallel ion source/mass analyzer/detector channels has been built to allow simultaneous high-throughput analysis of multiple samples. A multielement external chemical ionization/electron ionization source was coupled to a parallel array of CITs each of equal size (internal radius 2.5 mm), and the signal was recorded using an array of four miniature (2-mm inner diameter) electron multipliers. Using external electron ionization, the spectra of four separate samples were recorded simultaneously in real time using a four-channel preamplifier system and a data acquisition program written using LabVIEW software. These experiments mark the first demonstration of externally generated ions being successfully trapped in a miniature CIT mass analyzer. The instrument currently provides mass/charge range of approximately m/z 50-500. Average peak width is m/z 0.3, corresponding to a resolution of 1000 at m/z 300. The four-channel mass spectrometer is housed in a single vacuum manifold and operated with a single set of control electronics. The modular design of this instrument allows scale-up to many more channels of analysis for future applications in the areas of industrial process monitoring and combinatorial analysis and in the fields of proteomics and metabolomics.

Journal Article↗

Miniaturized and automated sample pretreatment for determination of PCBs in environmental aqueous samples using an on-line microporous membrane liquid-liquid extraction-gas chromatography system.

A new, fast, and automated sample pretreatment technique for determination of lipophilic organic compounds in aqueous samples has been developed and applied to the determination of polychlorinated biphenyls (PCBs) in environmental river water. It is based on miniaturized microporous membrane liquid-liquid extraction coupled on-line to gas chromatography (GC) with electron capture detection. The heart of the system that simultaneously connects the sample pretreatment step to the final GC analysis has been named the extracting syringe (ESy). The ESy carries a miniaturized membrane extraction card attached to an electrically and mechanically designed installment and is mounted directly over a GC injector for fully automated injection of the extract. A method was developed to extract 10 PCB congeners from 1-mL water samples (after addition of 40% acetonitrile) with an extraction time of 10 min. The optimized methodology showed good linearity (in the dynamic concentration range of 5 ng L(-)(1)-1 microg L(-)(1)), enrichment factors of 33-40 times, repeatable extractions (RSD 2-5%, n = 4), and low detection limits (2-3 ng L(-)(1)). Acetonitrile had to be added to the samples in order to overcome the influence of PCB adsorption on the repeatability of extraction and enrichment and to minimize the overall memory effect (OME). OME and carryover depended not only on the concentration of the organic solvent added to the sample and that used in the washing procedure but also on whether the extracting card was changed or not. When an optimized washing procedure was applied, the OME was approximately 0.2% at high concentrations (i.e., 1 microg L(-)(1)). When each extraction took place in a new extraction card, no OME was detected. Additionally, no significant adsorption onto glass surfaces or a matrix effect on extraction was noticed. The main features of this methodology are good extraction repeatability, low detection limits at short extraction time, and the unsurpassed characteristic of no detectable OME in the entire system when each sample is processed in a new card. The total consumption of organic (nonchlorinated) solvents is less than 5 mL per sample.

Journal Article↗

Mechanistic investigation of nanoparticle motion in pulsed voltage miniaturized electrical field flow fractionation device by in situ fluorescence imaging.

In our previous study, we reported a miniaturized electrical field flow fractionation device (micro-EFFF) that used a pulsed voltage (PV) to increase the effective electric field and, hence, improved the separation performance. In this work, we developed two micro-EFFFs with planar or segmented electrode design and investigated the particle movement in the flow channels under a PV. Numerical simulation was used to understand the electric field distribution in the micro-EFFFs. When the calculations for the micro-EFFF with a segmented electrode (segmented micro-EFFF) and the micro-EFFF with planar electrodes (planar micro-EFFF) are compared, a stronger electric field at the top electrode segments is found in the segmented micro-EFFF, with the strongest field at the edges of the electrode segments. Nanoparticle motion in both devices was in situ visualized by using a fluorescence microscope equipped with a CCD-camera. Results reveal that electrophoresis governs the nanoparticle movement in the planar micro-EFFF and dielectrophoresis dominates the movement in the segmented micro-EFFF. Two models are postulated to explain the experimental observations of the nanoparticle movement. The mechanistic understanding of controlling nanoparticle motion in a miniaturized environment will help the design and application of micro-EFFF for the separation of charged biomolecules (proteins and DNAs).

Journal Article↗

Measurement of ion swarm distribution functions in miniature low-temperature co-fired ceramic ion mobility spectrometer drift tubes.

Measurements of the performance of a miniature, portable 12-mm-diameter, 57-mm-length low-temperature cofired ceramic (LTCC) ion mobility spectrometer drift tube were undertaken to verify models of ion transport and determine the physical shape of the ion "swarms" in the LTCC tube. Simplified two-dimensional Gaussian models of ion swarm shape were fit to measured data to extract geometrical shape parameters. Results indicate that tube-transfer function effects that produce asymmetric ion swarms are minimized in the tube reducing temporal dispersion. Data are presented that illustrate the swarm shape as a function of gate time, electric field magnitude, and total charge in the ion swarm. Characterization and understanding of the ion transport mechanisms and effects that limit the resolution and other performance parameters of miniature IMS drift tubes is essential to the development of practical, robust, portable systems for "first responder" and homeland security missions.

Journal Article↗

Atomic absorption spectroscopy for mercury, automated by sequential injection and miniaturized in lab-on-valve system.

Sodium borohydride-based hydride generation was automated by using programmable flow within the lab-on-valve module. Mercury vapor, generated in the reaction mixture, was extracted in a gas/liquid separator. The gas-expansion separator was miniaturized and compared with the performance of a novel gas separator that exploits the combination of Venturi effect and reduced pressure. Cold vapor atomic spectroscopy was used as a model system, with detection of mercury by absorption at 254 nm and limit of detection of 9 microg of Hg/L, using 300 microL of sample and 100 microL of borohydride. This work introduces, for the first time, sequential injection technique for hydride generation, highlights advantages of using programmable flow, and outlines means for miniaturization of assays based on spectroscopy of volatile species.

Journal Article↗

Miniaturized ultrathin slab gel electrophoresis with thermal lens microscope detection and its application to fast genetic diagnosis.

A miniaturized ultrathin slab gel electrophoresis (MUSGE) apparatus was developed, and fast separation of DNA fragments was obtained using it. To obtain sufficient separation efficiency in a limited space, a discontinuous buffer system was used. In general, it is difficult to cast a discontinuous ultrathin slab gel of adequate quality. However, the miniaturized resolving gel could be cast by taking advantage of the "capillary phenomenon" of the ultrathin channel. A gradient plate was used to control the height of the resolving gel and to form a clear interface between the concentrating gel and the resolving gel. This method was used to cast multiple gels simultaneously and reproducibly. The gradient plate also facilitated sample introduction, which was carried out by using a micropipet. A 25-mm-long and 80-micron thick-resolving gel was used to separate the 100-base pair ladder DNA within 10 min. Bandwidth was reduced to 100-200 microns, thus improving the number of theoretical plates to 22,000, which was comparable to that in conventional slab gel electrophoresis even though the migration distance was reduced to 1/10. Satisfactory lane-to-lane reproducibility (RSD < 1.0%, n = 6) and gel-to-gel reproducibility (RSD < 2.7%, n = 4) were obtained. Finally, the MUSGE apparatus was successfully applied to get a rapid genetic diagnosis.

Apolipoproteins B↗

Characterization of oxygen transfer in miniature and lab-scale bubble column bioreactors and comparison of microbial growth performance based on constant k(L)a.

This work describes the engineering characterization of miniature (2 mL) and laboratory-scale (100 mL) bubble column bioreactors useful for the cultivation of microbial cells. These bioreactors were constructed of glass and used a range of sintered glass gas diffusers with differently sized pores to disperse humidified air within the liquid biomedium. The effect of the pressure of this supplied air on the breakthrough point for gas diffusers with different pore sizes was examined and could be predicted using the Laplace-Young equation. The influence of the superficial gas velocity (u(g)) on the volumetric mass transfer coefficient (k(L)a) was determined, and values of up to 0.09 s(-1) were observed in this work. Two modeling approaches were considered in order to predict and provide comparison criteria. The first related the volumetric power consumption (P/V) to the k(L)a and a good correlation was obtained for differently sized reactors with a given pore size, but this correlation was not satisfactory for bubble columns with different gas diffusers. Values for P/V ranged from about 10 to 400 W.m(-3). Second, a model was developed predicting bubble size (d(b)), bubble rising velocity (u(b)), gas hold-up (phi), liquid side mass transfer coefficient (k(L)), and thus the k(L)a using established theory and empirical correlations. Good agreement was found with our experimental data at different scales and pore sizes. Values for d(b) varied from 0.1 to 0.6 mm, and k(L) values between 1.7 and 9.8 x 10(-4) m.s(-1) were determined. Several E. coli cultivations were performed in the miniature bubble column at low and high k(L)a values, and the results were compared to those from a conventional stirred tank operated under identical k(L)a values. Results from the two systems were similar in terms of biomass growth rate and carbon source utilization.

Bioreactors↗

A miniature bioreactor for sensing toxicity using recombinant bioluminescent Escherichia coli cells.

A miniature bioreactor was fabricated as a contactor between biosensing cells and toxic materials. This miniature bioreactor (58 mL working volume) showed performance similar to that of a conventional bioreactor, as well as the advantages of easy installation, facile operation, and small medium requirements during long-term continuous operation. A performance evaluation measured the response to ethanol in continuous operation by using a recombinant bioluminescent Escherichia coli strain. Continuous cultures were repeatedly induced by the ethanol challenge. Steady-state cell concentrations (OD) were found to be decreased, the induced specific bioluminescence (SBL) peak value was found to be increased, and the peak response time, which is the time constant of this continuous monitoring system, was found to be decreased with increasing dilution rate. Finally on- and off-line bioluminescence monitoring was shown to be reliable, suggesting that this system is suitable for applications such as monitoring the influent and effluent streams of waste water biotreatment plants.

Biosensing Techniques↗

Inhibition of beta-amyloid fibrillization by directed evolution of a beta-sheet presenting miniature protein.

We describe the directed evolution of a miniature beta-sheet protein for targeting beta-amyloid oligomers implicated in Alzheimer's disease. Circular dichroism spectroscopy, thermal denaturation experiments, and immunoglobulin binding assays established that our beta-amyloid-targeted miniature protein, TJ10, presents a well-folded thermostable beta-sheet. TJ10 was found to prevent beta-amyloid fibrillization at stoichiometric concentrations and was also an effective inhibitor at substoichiometric concentrations. Thus our results provide a new and potent beta-sheet chemical template for effectively targeting beta-amyloid while also demonstrating a general strategy for targeting proteins implicated in other amyloidogenic diseases.

Amino Acid Sequence↗

Aroma-active compounds of miniature beefsteakplant (Mosla dianthera Maxim).

Volatile flavor compounds of miniature beefsteakplant (Mosla dianthera Maxim.) from Vietnam were analyzed through gas chromatography-mass spectrometry-olfactometry (GC-MS-O). Sixty-two compounds were identified by GC-MS. Of these, (+/-)-carvone and (+/-)-limonene were the most abundant, followed by (Z)-limonene oxide, beta-caryophyllene, and alpha-humulene. Twenty aroma-active compounds were detected by aroma extract dilution analysis conducted on two GC columns of different polarities (DB-5MS and DB-Wax). The most intense aroma-active compounds were linalool (floral/sweet/lemon), (-)-carvone (spearminty), and 1-octen-3-one (mushroom/earthy). Other predominant aroma-active compounds included (Z)-3-hexenol (grassy/leafy/metallic), (Z)-limonene oxide (lemon/floral), myrcene (plastic/sweet), (+)-limonene (orange/lemon), alpha-thujene (soy sauce/grassy), and (Z)-dihydrocarvone (spearminty/pepperminty). On the basis of the aroma characteristics and intensity, it was concluded that (-)-carvone was responsible for the characteristic aroma of miniature beefsteakplant.

Gas Chromatography-Mass Spectrometry↗