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Sampling small volumes of ambient ammonia using a miniaturized gas sampler.

The development of a gas sampler for a miniaturized ambient ammonia detector is described. A micromachined channel system is realized in glass and silicon using powder blasting and anodic bonding. The analyte gas is directly mixed with purified water, dissolving the ammonia that will dissociate into ammonium ions. Carrier gas bubbles are subsequently removed from the liquid stream through a venting hole sealed with a microporous water repellent PTFE membrane. A flow restrictor is placed at the outlet of the sampler to create a small overpressure underneath the membrane, enabling the gas to leave through the membrane. Experiments with a gas flow of 1 ml min(-1), containing ammonia concentrations ranging from 9.4 ppm to 0.6 ppm in a nitrogen carrier flow have been carried out, at a water flow of 20 microl min(-1). The ammonium concentration in the sample solution is measured with an electrolyte conductivity detector. The measured values correspond with the concentration calculated from the initial ammonia concentration in the analyte gas, the fifty times concentration enhancement due to the gas-liquid volume difference and the theoretical dissociation equilibrium as a function of the resulting pH.

Journal Article↗

Solid phase micro-extraction in a miniature ion trap mass spectrometer.

Fiber introduction mass spectrometry (FIMS), a variation of solid-phase microextraction (SPME) and membrane introduction mass spectrometry (MIMS), is employed with a miniature mass spectrometer. The inlet system, constructed of commercially available vacuum parts, allows the direct introduction of the SPME needle vacuum chamber into the mass spectrometer. Thermal desorption of the analyte from the poly(dimethylsiloxane) (PDMS) coated fiber was achieved with a built in nichrome heater, followed by electron ionization of the analytes internal to the cylindrical ion trap (CIT). The system has been tested with several volatile organic compounds (VOC) in air and to analyze the headspace over aqueous solutions, with limits of detection in the low ppb range. The signal rise (10-90%) and fall (90-10%) times for the system ranged from 0.1 to 1 s (rise) and 1.2 to 6 s (fall) using heated desorption. In addition, this method has been applied to quantitation of toluene in benzene, toluene, xylene (BTX) mixtures in water and gasoline. This simple and rapid analysis method, coupled to a portable mass spectrometer, has been shown to provide a robust, simple, rapid, reproducible, accurate and sensitive (low ppb range) fieldable approach to the effective in situ analysis of VOC in various matrices.

Ions↗

Nucleophilic and general acid catalysis at physiological pH by a designed miniature esterase.

A 31-residue peptide (Art-Est) was designed to catalyse the hydrolysis of p-nitrophenyl esters through histidine catalysis on the solvent exposed face of the alpha-helix of bovine pancreatic polypeptide. NMR spectroscopy indicated that Art-Est adopted a stable 3-dimensional structure in solution. Art-Est was an efficient catalyst with second order rate constants of up to 0.050 M(-1) s(-1). The activity of Art-Est was a consequence of the increased nucleophilicity of His-22, which had a reduced pK(a) value of 5.5 as a consequence of its interaction with His-18 and the positively charged Arg-25 and Arg-26. Mass spectrometry and NMR spectroscopy confirmed that the Art-Est catalysed hydrolysis of p-nitrophenyl esters proceeded through an acyl-enzyme intermediate. A solvent kinetic isotope effect of 1.8 indicated that the transition state preceding the acyl intermediate was stabilised through interaction with the protonated side-chain of His-18 and indicated a reaction mechanism similar to that generally observed for natural esterases. The involvement in the reaction of two histidine residues with different pK(a) values led to a bell-shaped dependence of the reaction rate on the pH of the solution. The catalytic behaviour of Art-Est indicated that designed miniature enzymes can act in a transparent mechanism based fashion with enzyme-like behaviour through the interplay of several amino acid residues.

Amino Acid Sequence↗

Flow-based and sieving matrix-free DNA differentiation by a miniaturized field flow fractionation device.

DNA separation is typically done by gel electrophoresis based on its charge property. In our previous work, we reported that dielectrophoresis could be used to manipulate polystyrene nanoparticles' motion by using a miniaturized electrical field flow fractionation device (micro-EFFF) with a segmented electrode operated under a pulsed voltage (PV). In this work, we report the manipulation and separation of DNA molecules using the micro-EFFF. DNA motion was in situ visualized inside the device. Results revealed that dielectrophoresis governed DNA motion, which was strongly correlated with the pulse frequency but not the duty cycle of a PV. A longer retention time of DNA molecules was measured under a PV. The retention time increased with the length of DNA molecules. As the micro-EFFF is flow-based and sieving-matrix-free, it has a potential to be applied to sample preparation in a micrototal analysis system or when fractionated molecules are needed for downstream analysis.

DNA, Bacterial↗

Novel integrated paired emitter-detector diode (PEDD) as a miniaturized photometric detector in HPLC.

A novel low power, low cost, highly sensitive, miniaturized light emitting diode (LED) based flow detector has been used as optical detector for the detection of sample components in high performance liquid chromatography (HPLC). This colorimetric detector employs two LEDs, one operating in normal mode as a light source and the other is reverse biased to work as a light detector. Instead of measuring the photocurrent directly, a simple timer circuit is used to measure the time taken for the photocurrent generated by the emitter LED (lambda(max) 500 nm) to discharge the detector LED (lambda(max) 621 nm) from 5 V (logic 1) to 1.7 V (logic 0) to give digital output directly without using an A/D converter. Employing a post-column reagent method, a Nucleosil 100-7 column (functionalized with iminodiacetic acid (IDA) groups) was used to separate a mixture of transition metal complexes, manganese(II) and cobalt(II) in 4-(2-pyridylazo)-resorcinol (PAR). All optical measurements were taken by using both the in-built HPLC variable wavelength detector and the proposed paired-emitter-detector-diode (PEDD) optical detector configured in-line for data comparison. The concentration range investigated using the PEDD was found to give a linear response to the Mn(II) and Co(II) PAR complexes. The effects of flow rate and emitter LED light source intensity were investigated. Under optimised conditions the PEDD detector offered a linear range of 0.9-100 microM and LOD of 0.09 microM for Mn-PAR complex. A linear range of 0.2-100 microM and LOD of 0.09 microM for Co-PAR complex was achieved.

Journal Article↗

Lipid accumulation in liver, spleen, lungs and kidneys of miniature-pigs after chloroquine treatment.

Chronic chloroquine treatment of type-Göttingen miniature-pigs induced lipid accumulation in the liver, spleen, lungs and kidneys. The lipid analyses showed marked quantitative and qualitative differences between the organs. In the liver the lipids affected most were cholesteryl esters and glucosylceramides, which were increased at the most 20 times. Cholesterol and ganglioside concentrations were also increased, though less markedly. The concentration of acidic phospholipids was slightly increased but that of the neutral phospholipids was unaffected. There was a considerable inter-individual variation in the lipid changes. Spleen and lung showed significant increases of all the major lipids. Glucosylceramide was increased more than the other lipids, namely 6-fold in the spleen and 10-fold in the lung. The concentration of acidic phospholipids as well as that of gangliosides was increased by 50% in the spleen and by 100% in the lung. The organ affected least was the kidney, in which only the glycolipids, both acidic and neutral, were significantly increased. Common to all the organs of the chloroquine-treated pigs was the large increase of glucosylceramide, ganglioside CM2 and bis(monoacylglyceryl)phosphate. The ganglioside increase affected all the individual gangliosides and, except for the increased proportion of ganglioside GM2, there were not remarkable changes in the ganglioside pattern in any of the organs.

Animals↗

Experimental hyperthyroidism does not induce hepatic insulin resistance in the miniature pig.

The effect of hypo- and hyper-thyroidism on insulin-mediated alterations in tracer-determined glucose kinetics and the arterial concentration of gluconeogenic precursors were investigated in 24 h-starved conscious unrestrained miniature pigs. Hyperinsulinaemia (about 40 microunits/ml) decreased blood glucose and, transiently, glucose output at unaltered glucose utilization in all thyroid states: this effect was pronounced in hyperthyroid (-50%) and less in hypothyroid pigs (-25%) compared with euthyroid controls (-35%). We conclude that moderate experimental hyperthyroidism does not induce hepatic insulin resistance, whereas hypothyroidism slightly impairs insulin action with respect to the regulation of glucose output.

Animals↗

A miniature tooth-borne distractor for the alignment of ankylosed teeth.

The ankylosis of a tooth is one of the most difficult clinical problems that an orthodontist faces. In the literature, the treatment protocols for ankylosed teeth are still insufficient and questionable when considering gingival esthetics and conservation of bone health. The purpose of this report is to evaluate and discuss the effects of a newly designed miniature tooth distractor (MTD), which can be used with infrapositioned ankylosed teeth. Two cases with vertically malpositioned incisors were treated using the MTD, and this device was evaluated and compared with the distraction appliances used before in the literature. In conclusion, it was found to be efficient with its small dimensions, ease of application and removal, ease of activation, buccolingual control, and patient tolerance.

Adolescent↗

Using the miniature linguistic system in teaching syntax: two case studies.

Two language-delayed children received comprehension instruction on preposition-object phrases using a miniature linguistic system format to structure generalization. Results indicated generalization not only to untreated two-word combinations, but also to comprehension of four-word phrases. In addition, there was some generalization of production of two- and four-word structures.

Child, Preschool↗

Acquisition of a verb-subject-object miniature linguistic system by adults.

Two experiments are reported. In the first, ten college-aged adults were exposed to a miniature linguistic system (MLS) and the referents it encoded. The referents included three shapes which functioned as subjects in the MLS, three shapes which functioned as objects, and three actions which functioned as verbs. Nine nonsense syllables were assigned to the referents. The MLS allowed only sentences of the form, verb-subject-object, to describe a subject-shape acting in a specific manner on an object-shape. Videotaped presentations of the shape and action referents and simultaneous auditory presentation of the appropriate MLS sentence were used during teaching trials. Testing trials containing no auditory presentations were alternated with the teaching trials. During testing trials, subjects were required to write MLS sentences to describe the shape-action referents presented. The data gathered during testing trials allowed the acquisition of each lexical item, each word-class, and the word-class ordering rule to be examined. The results of Experiment I showed that the actions were learned more quickly than either the subject-shapes or the object-shapes. Experiment II investigated possible reasons for the verb-learning advantage noted in Experiment I. Twelve college-aged adults learned shape names and action names as single word responses. The results of Experiment II suggest that the verb-learning advantage noted in Experiment I was due, at least in part, to a lexical learning effect rather than a syntactical learning effect, nonsense syllable mediation, or word position. Error analyses of Experiment I data support this conclusion. Implications of the findings and the experimental technique for language remediation are discussed.

Adult↗

Detecting nasalization using a low-cost miniature accelerometer.

A low-cost miniature accelerometer attached externally on the nose is as effective in providing an indication of nasalization as a more expensive accelerometer used in previous research. The more limited bandwidth of this low-cost accelerometer does not restrict its ability to monitor nasalization because the vibratory energy of interest is below 2,000 Hz. The optimal placement of the accelerometer on the nose is over the lateral nasal cartilage, a position along the upper side of the nose, just in front of the nasal bone.

Adult↗

Miniature language system acquisition by children with different learning proficiencies.

Sixteen children, aged 7:8 to 9:10, learned two miniature languages while playing a communication game. Both languages expressed Action (Agent, Patient) meanings and incorporated a Patient suffix. They differed in word order: VSO (Language I) versus SOV (Language II). Children found the SOV language easier; they also made more suffix errors and fewer word order errors in this language. The results suggest that the perceptual salience of an utterance-final particle may hinder grammatical analysis, at least if capacity limits and perseverative learning strategies intervene.

Attention↗

BDNF reduces miniature inhibitory postsynaptic currents by rapid downregulation of GABA(A) receptor surface expression.

Changes in neurotransmitter receptor density at the synapse have been proposed as a mechanism underlying synaptic plasticity. Neurotrophic factors are known to influence synaptic strength rapidly. In the present study, we found that brain-derived neurotrophic factor (BDNF) acts postsynaptically to reduce gamma-aminobutyric acid (GABA)-ergic function. Using primary cultures of rat hippocampal neurons, we investigated the effects of BDNF on GABAergic miniature inhibitory postsynaptic currents (mIPSCs) and on the localization of GABAA receptors. Application of BDNF (100 ng/mL) led within minutes to a marked reduction (33.5%) of mIPSC amplitudes in 50% of neurons, recorded in the whole-cell patch-clamp mode, leaving frequency and decay kinetics unaffected. This effect was blocked by the protein kinase inhibitor K252a, which binds with high affinity to trkB receptors. Immunofluorescence staining with an antibody against trkB revealed that about 70% of cultured hippocampal pyramidal cells express trkB. In dual labelling experiments, use of neurobiotin injections to label the recorded cells revealed that all cells responsive to BDNF were immunopositive for trkB. Treatment of cultures with BDNF reduced the immunoreactivity for the GABAA receptor subunits-alpha2, -beta2,3 and -gamma2 in the majority of neurons. This effect was detectable after 15 min and lasted at least 12 h. Neurotrophin-4 (NT-4), but not neurotrophin-3 (NT-3), also reduced GABAA receptor immunoreactivity, supporting the proposal that this effect is mediated by trkB. Altogether the results suggest that exposure to BDNF induces a rapid reduction in postsynaptic GABAA receptor number that is responsible for the decline in GABAergic mIPSC amplitudes.

Animals↗

A linkage map of 243 DNA markers in an intercross of Göttingen miniature and Meishan pigs.

A resource family of pigs has been constructed by using a boar of Göttingen miniature pig and two sows of Meishan pig as parents. In the construction of the family, two F1 males and 18 F1 females were intercrossed to generate 143 F2 offspring. The members of the family were genotyped using 243 genetic markers including 26 markers developed in our laboratory in order to generate a linkage map of markers for use in detecting quantitative trait loci (QTLs) in the family. The markers consisted of 237 microsatellites, five PRE-1 markers, and one RFLP marker. The linkage map was revealed to cover all 18 autosomes and the X chromosome; and the total length of the sex-averaged linkage map was calculated to be 2561.9 CM. Four out of the 26 markers developed in our laboratory exended the current linkage map at the termini of chromosomes 1p, 5p, 11p, and Xq. The linkage maps of all the chromosomes except for chromosome 1 were found to be longer in females than in males. Concerning chromosome 1, the length of the linkage map showed no difference between females and males, which was attributed to low recombination rates between markers localized in the centromeric region in females. The average ratio of female-to-male recombination was calculated to be 1.55.

Animals↗

DEC: a new miniature inverted-repeat transposable element from the genome of the beetle Tenebrio molitor.

In this paper we describe a novel family of miniature inverted-repeat transposable elements (MITEs), named DEC, isolated from the genome of the beetle Tenebrio molitor. These elements are highly reiterated and their number is estimated to be around 3500 per haploid genome. Two of them have been isolated and the two sequences are 84% identical. Like other MITEs, they are characterized by their small size, their A + T richness, the presence of terminal inverted repeats and the absence of open reading frames. These data suggest that MITEs are probably widely distributed in arthropods.

Animals↗

Central vagal stimulation evokes gastric volume changes in mice: a novel technique using a miniaturized barostat.

We have developed a novel technique to measure gastric volume in vivo in mice; this will be invaluable for revealing gastric alterations in genetically modified mice models, thus expanding our understanding of the mechanisms underlying functional disorders. Experimental data on gastric tone currently available has focused on rats using isovolumetric techniques to measure pressure changes, whereas clinical studies use barostatic techniques to measure volume changes. For better translational approaches, we assessed the feasibility of using a miniaturized barostat to measure gastric volume changes in urethane-anaesthetized and unanaesthetized-decerebrate mice. Additionally, we assessed whether central vagal stimulation alters gastric volume in urethane-anaesthetized mice. Nitric oxide donor sodium nitroprusside (1mg kg-1 i.p.) increased gastric volume (+134 +/- 20 microL), whereas the cholinergic agonist carbachol (3 microg kg-1 i.p.) decreased gastric volume (-153 +/- 20 microL). Similar responses were obtained in urethane-anaesthetized and unanaesthetized-decerebrate animals. Microinjection of L-glutamate (25 nmol) into dorsal motor nucleus of the vagus (DMV) altered gastric volume; microinjection into rostral DMV led to gastric contraction (-83 +/- 18 microL) while stimulation of caudal DMV resulted in gastric relaxation (+95 +/- 16 microL). This reveals a functional organization of DMV in mice. This study validates barostatic techniques for application to mice. An understanding of gastric contractility and tone is clinically relevant as impaired gastric accommodation reflex may be an underlying cause of functional dyspepsia.

Anesthetics, Intravenous↗

Presence of miniature inverted-repeat transposable elements (MITEs) in the genome of Arabidopsis thaliana: characterisation of the Emigrant family of elements.

Although the genome of Arabidopsis thaliana has a small amount of repetitive DNA, it contains representatives of most classes of mobile elements. However, to date, no miniature inverted-repeat transposable element (MITE) has been described in this plant. Here, we describe a new family of repeated sequences that we have named Emigrant, which are dispersed in the genome of Arabidopsis and fulfil all the requirements of MITEs. These sequences are short, AT-rich, have terminal inverted repeats (TIRs), and do not seem to have any coding capacity. Evidence for the mobility of Emigrant elements has been obtained from the absence of one of these elements in a specific Arabidopsis ecotype. Emigrant is also present in the genome of different Brassicae and its TIRs are 74% identical to those of Wujin elements, a recently described family of MITEs from the yellow fever mosquito Aedes aegypti.

Arabidopsis↗

Altering the concentration of GABA in the synaptic cleft potentiates miniature IPSCs in rat occipital cortex.

We have tested the effect of dextran (40 kDa, 5%) on miniature IPSCs (mIPSCs) recorded in layer V cortical pyramidal cells. This compound increases the amplitude of mIPSCs at room and physiological temperatures by 15%, leaving their duration unaffected at room temperature and slightly increased at physiological temperature. The amplitude increase is attributable to an increase in the number of receptors bound by GABA during synaptic transmission, as shown by the occlusion between the effects of dextran and zolpidem on mIPSC amplitude at room temperature. As dextran presumably enhances the concentration and dwell time of GABA in the synaptic cleft, these results demonstrate that the postsynaptic GABAA receptors are not saturated at room and physiological temperatures.

Animals↗