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Potentiometric detection of carboxylic acids, phosphate esters, and nucleotides in liquid chromatography using anion-selective coated-wire electrodes.

An all-solid-state ion-selective membrane electrode incorporating a lipophilic anion exchanger was used in a flow-through potentiometric detector for the LC determination of organic anions of biological interest. Different metabolic intermediates (mono-, di-. and tricarboxylic acids, sugar phosphates, and nucleotides) were detected sensitively after separation on a pellicular anion-exchange chromatographic column. The electrode was coated by directly casting the electroactive mixture on a glassy carbon support of 3 mm diameter and used in a wall-jet-type flow cell. The analysis conditions were optimized to obtain both efficient separation and sensitive detection. Calibration curves showed a logarithmic dependence on the injected concentration for concentrations higher than 5.0 x 10(-5) M and a linear dependence for injected concentrations below this value. Under isocratic conditions, detection limits of 5.0 x 10(-7) M (25 pmol) were attained when a sodium hydroxide solution was used as an eluent. No suppressor system was needed in this case. The relative standard deviation for consecutive injections was 0.3% (n = 15), and the electrode lifetime was at least 2 months. The utility of potentiometric detection is further demonstrated in a gradient elution separation for single-run analysis of a synthetic mixture of biochemical compounds containing carboxylic acids, phosphate esters, and nucleotides.

Algorithms↗

Effect of orifice-area reduction on flow characteristics during injection through spinal needles.

A reduction in hole size for certain side-port spinal needles has been advocated in recent reports. While the influence of orifice-area reduction on the aspiration capability of the needle has been studied, the influence on the anaesthetic delivery properties is relatively unknown. As a first step in understanding the effects of hole-size reduction on anaesthetic distribution within the subarachnoid space, we studied flows emanating from isolated needles using computer simulations. Following validation of the numerical model using experimental particle visualisation, trajectories of anaesthetic particles injected through 25 G Whitacre needles of various orifice areas were computed and used to determine the orientation and rate of spread of the anaesthetic jet exiting the needle. Two factors impacting the concentration distribution were observed: the rate of spread of the anaesthetic jet increases markedly with decreasing orifice area and the jet alignment shifts toward perpendicular to the needle axis.

Anesthesia, Spinal↗

[Inferior vena caval angiography: a new angiographic method for evaluating tricuspid regurgitation].

It is well known that right ventriculography has unavoidable disadvantages as a method for diagnosing tricuspid regurgitation. In this study, inferior vena caval angiography (IVC angiography) was tested as a new method for quantitatively diagnosing tricuspid regurgitation. With this method, no catheter passes through the tricuspid valve, and only a small amount (10 ml) of contrast material injected into the upper portion of the inferior vena cava visualizes the entire right atrium, and tricuspid regurgitation is manifested by turbulence or a negative jet in the right atrium. With respect to the degree, tricuspid regurgitation was graded as absent (0), mild (1+), moderate (2+) and severe (3+) using the criteria shown in Fig. 1. Mild tricuspid regurgitation was diagnosed when systolic turbulence was observed in the right atrium and did not reach the right atrial wall. Moderate tricuspid regurgitation was diagnosed when systolic turbulence reached the right atrial wall. Severe tricuspid regurgitation was diagnosed when systolic turbulence entered the inferior vena cava. Sixty-four patients with valvular heart disease and four having coronary heart disease were studied using IVC angiography and pulsed Doppler echocardiography. Using Doppler, the severity of tricuspid regurgitation was determined according to the distribution of the regurgitant signal in the right atrium. The degree of tricuspid regurgitation by IVC angiography correlated well with that by Doppler. All patients with severe (3+) regurgitation and 15 of 22 patients with moderate (2+) regurgitation required surgery, but all with no (0) regurgitation and 12 of 14 with only mild (1+) regurgitation required no surgical correction of the tricuspid valve.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiography↗

Thermal chip fabrication with arrays of sensors and heaters for micro-scale impingement cooling heat transfer analysis and measurements.

The design and fabrication for a thermal chip with an array of temperature sensors and heaters for study of micro-jet impingement cooling heat transfer process are presented. This thermal chip can minimize the heat loss from the system to the ambient and provide a uniform heat flux along the wall, thus local heat transfer processes along the wall can be measured and obtained. The fabrication procedure presented can reach a chip yield of 100%, and every one of the sensors and heaters on the chip is in good condition. In addition, micro-jet impingement cooling experiments are performed to obtain the micro-scale local heat transfer Nusselt number along the wall. Flow visualization for the micro-impinging jet is also made. The experimental results indicate that both the micro-scale impinging jet flow structure and the heat transfer process along the wall is significantly different from the case of large-scale jet impingement cooling process.

Cold Temperature↗

A continuous-flow capillary mixing method to monitor reactions on the microsecond time scale.

A continuous-flow capillary mixing apparatus, based on the original design of Regenfuss et al. (Regenfuss, P., R. M. Clegg, M. J. Fulwyler, F. J. Barrantes, and T. M. Jovin. 1985. Rev. Sci. Instrum. 56:283-290), has been developed with significant advances in mixer design, detection method and data analysis. To overcome the problems associated with the free-flowing jet used for observation in the original design (instability, optical artifacts due to scattering, poor definition of the geometry), the solution emerging from the capillary is injected directly into a flow-cell joined to the tip of the outer capillary via a ground-glass joint. The reaction kinetics are followed by measuring fluorescence versus distance downstream from the mixer, using an Hg(Xe) arc lamp for excitation and a digital camera with a UV-sensitized CCD detector for detection. Test reactions involving fluorescent dyes indicate that mixing is completed within 15 micros of its initiation and that the dead time of the measurement is 45 +/- 5 micros, which represents a >30-fold improvement in time resolution over conventional stopped-flow instruments. The high sensitivity and linearity of the CCD camera have been instrumental in obtaining artifact-free kinetic data over the time window from approximately 45 micros to a few milliseconds with signal-to-noise levels comparable to those of conventional methods. The scope of the method is discussed and illustrated with an example of a protein folding reaction.

Biophysics↗

Acute mental stress but not enforced muscle activity transiently increases natural cytotoxicity in spontaneously hypertensive rats.

The influence of acute mental stress and the effect of electrically induced skeletal muscle contractions on natural cytotoxicity in vivo was investigated in spontaneously hypertensive rats Natural cytotoxicity in vivo was measured as the clearance of injected 51Cr-labelled YAC-1 lymphoma cells from the lungs, which are specifically lysed by natural killer cells. The mental stress consisted of an air jet directed towards the animals in their cage for 25 min. During the mental stress there was a significant increase in natural cytotoxicity. Thus, retained radioactivity in the lungs was decreased to 74 +/- 6% of the control levels which was set to 100% (P < 0.01). This augmentation of YAC-1-cell clearance could be blocked with the beta-adrenergic receptor antagonist Timolol. Two hours after termination of the air stress, in vivo cytotoxicity had returned to control levels. In contrast, acute physical stress, consisting of electrically induced muscle contractions for 60 min, had no significant effects on in vivo cytotoxicity, either during the stimulation or 1, 2 or 24 h after the stimulation. Further, significantly increased plasma levels of adrenaline were seen after the air jet stress, but not after muscle stimulation. There were no significant changes in plasma noradrenaline levels either after air stress or muscle stimulation. These results indicate that changes in in vivo cytotoxicity after mild mental stress are dependent on increased plasma catecholamine levels while acute physical stress without changes in catecholamine levels, does not influence in vivo cytotoxicity.

Animals↗

Sample flow switching techniques on microfluidic chips.

This paper presents an experimental investigation into electrokinetically focused flow injection for bio-analytical applications. A novel microfluidic device for microfluidic sample handling is presented. The microfluidic chip is fabricated on glass substrates using conventional photolithographic and chemical etching processes and is bonded using a high-temperature fusion method. The proposed valve-less device is capable not only of directing a single sample flow to a specified output port, but also of driving multiple samples to separate outlet channels or even to a single outlet to facilitate sample mixing. The experimental results confirm that the sample flow can be electrokinetically pre-focused into a narrow stream and guided to the desired outlet port by means of a simple control voltage model. The microchip presented within this paper has considerable potential for use in a variety of applications, including high-throughput chemical analysis, cell fusion, fraction collection, sample mixing, and many other applications within the micro-total-analysis systems field.

Biosensing Techniques↗

Field trials of a very potent rabies DNA vaccine which induced long lasting virus neutralizing antibodies and protection in dogs in experimental conditions.

Kinetics of antibody responses and protection against rabies were investigated after injection of a single dose of rabies DNA vaccine and compared to those induced by one or two injections of cell culture-derived vaccine in dogs issued from the common local breed and reared in experimental conditions. Rabies DNA vaccine administered intradermally by a jet injector in the inner face of the ear was by far more efficient in inducing long lasting high titers of virus neutralizing antibodies compared to cell culture vaccine Rabisin administered either subcutaneously or intramuscularly. Four years after vaccine administration of either DNA or cell culture-derived rabies vaccines, full protection against a rabies peripheral challenge was achieved. Vaccine trials targeting dogs living in field conditions in Tunisia further established that rabies DNA-based vaccination induced a stronger induction of virus neutralizing antibodies compared to Rabisin. This report shows for the first time that DNA vaccination could be more efficient under experimental or field conditions in large size mammals than the best commercially available cell culture-derived vaccine. This improvement will hopefully allow a better rabies control in developing countries by using a more efficient vaccination with fewer doses and targeting all categories of dogs.

Animals↗

Potential for cross-contamination from use of a needleless injector.

BACKGROUND: Medical devices that are used on patients in fields containing potentially infectious body fluids can become contaminated and transmit infectious agents to other sites on the patient or to other patients if the devices are not properly cleaned and decontaminated after use on each patient treatment site. One such device is the needleless or jet injector, which is widely used in medicine and dentistry to deliver local anesthetic in procedures such as bone marrow aspirations, lumbar punctures, and cutaneous and intraoral injections. This study was conducted to determine whether cross-contamination can occur on in vitro reuse of a needleless injector and whether a manufacturer's recommended method of injector decontamination (ie, immersion sterilization) is effective in the prevention of cross-contamination. METHODS: The study was performed with new autoclaved injectors, fluorescein dye, and Streptococcus crista (the bacteria commonly found in saliva) in the field of use to determine whether these devices can become contaminated during use and carry over the contamination to other sites during immediate reuse. RESULTS: Fluorescein dye and bacteria tests with the needleless injectors showed that contamination or carryover does occur. It appeared to reduced to a minimum when a autoclaved, sterile rubber cap used over the head of the device during injection was replaced between each use, although replacement of the rubber cap alone did not prevent carryover. Immersion of the head of the injector in a 2% glutaraldehyde solution for 30 minutes followed by a sterile water rinse and the replacement of the rubber cap with a sterile cap between uses was shown to curtail bacterial growth and prevent cross-contamination on immediate reuse of the device. CONCLUSION: This study demonstrated that needleless injectors become contaminated during in vitro use and direct contact with contaminated surfaces and that needless injectors carry over the contamination to subsequent sites of release. The replacement of the injector's rubber cap with a new one after initial discharge or the removal of an exposed rubber cap and immersion of the head of the injector in 2% glutaraldehyde followed by a rinse of the head in sterile water, as recommended by one injector manufacturer, can minimize or eliminate the carryover.

Contrast Media↗

Melatonin and jet lag syndrome: experimental model and clinical implications.

What is the effect of melatonin on jet lag syndrome? Jet lag desynchronizes the internal sleep-wakefulness cycle with the environmental light/dark cycle. Advance (but not delay) of light onset is known to abolish pineal N-acetyltransferase activity and urine excretion of 6-sulphatoxymelatonin. Measurements of pineal serotonin, the substrate of melatonin biosynthesis; N-acetylserotonin (NAS), the immediate melatonin precursor; and melatonin (high-performance liquid chromatography-fluorimetric method) in the animal (rat) model of jet lag revealed that prolonged delay of dark-phase onset disrupted the rhytms in comparable ways as the advance of light-phase onset. Advance of dark phase onset resulted in less severe disturbances of rhythms as compared with the advance of light phase onset. Melatonin, but not NAS, injections at the beginning of a new dark period accelerated recovery of NAS and melatonin, but not serotonin, rhythms. Spontaneously hypertensive rats were more sensitive to advance of light onset and less responsive to melatonin injections than normotensive rats. NAS and methylene blue, an inhibitor of monoamine oxidase A, attenuated light-induced disruption of NAS but not melatonin rhythms. We draw the following conclusions from our data: the beginning of the dark period may be preferable to the beginning of light period as the arrival time on eastward flights; the efficacy of melatonin in alleviating jet lag may be enhanced by administering it before, during and after rapid transition through time zones; and hypertension may exaggerate jet lag syndrome.

Animals↗

[Comparative study of high-frequency jet ventilation in 4 types of patients undergoing laryngeal microsurgery].

OBJECTIVE: To study high frequency jet ventilation (HFJV) in patients with chronic obstructive pulmonary disease (COPD) who are obese or who have tracheal or laryngeal stenosis, conditions which are often found in patients undergoing microsurgery of the larynx (MSL), comparing the results with those for a group of controls, patients with no stenosis who were also undergoing MSL. PATIENTS AND METHOD: Eighty patients were distributed in four groups as follows: those meeting the criteria for a diagnosis of COPD (n = 20), those who were overweight (n = 24), those with stenosis over 50% of the laryngeal opening (stenosis group, n = 10) and those with no associated pathology (control group, n = 26). HFJV was administered through a 2.2 mm internal diameter orotracheal injection catheter, using an Ergojet CVT (Temel, S.A.). The ventilatory protocol was as follows: rate 100 breaths/min, inspiratory time 30%, generator pressure (GP) 2.2 to 3.3 kg/cm2 and FiO2 70% to 90%. We analyzed the GP administered, jet volume (Vjet) delivered, maximum (PAWmax) and minimum (PAWmin) airway pressures, oxygen hemoglobin saturation (SpO2), partial pressures of O2 (pO2) and CO2 (pCO2) in arterial blood, and end-tidal pressure of CO2 (PETCO2) at baseline and 10 and 20 min after the start of HFJV. Because monitoring was invasive, the study was designed for a small series of patients and we believe it should not be generalized to include all patients undergoing MSL under normal conditions. RESULTS: HFJV had to be abandoned and conventional ventilation used in 4 patients (2 in the COPD group and 2 in the obese group). Ventilation was judged adequate in the remaining patients, with the observations that in the COPD group, pO2 levels were lower than in the control group at the 10 min readings and pCO2 levels were higher at both the 10 and 20 min readings. PAW levels were higher throughout the procedure in both the COPD and stenosis groups. In the obese patients, pCO2 was higher at both the 10 and 20 min recordings. CONCLUSION: HFJV provides effective ventilation for most patients undergoing MSL, even if they are obese or have COPD or laryngeal stenosis.

Adult↗

Postsynaptic potentials evoked in ventrobasal thalamus neurones by natural sensory stimuli.

Intracellular recordings were made in the ventrobasal thalamus of rats anaesthetised with urethane. Postsynaptic responses were evoked by stimulation of the peripheral receptive field with an air jet of 10 ms duration. The postsynaptic response typically consisted of an excitatory postsynaptic potential (EPSP)/inhibitory postsynaptic potential (IPSP) sequence with one or more evoked action potentials. Injection of hyperpolarizing current pulses appeared to increase the EPSP amplitude, whereas depolarising current pulses caused a reduction in EPSP amplitude. These changes in EPSP amplitude were however obscured by the presence of IPSPs and a slow potential similar to a low-threshold Ca2+ spike (LTS).

Animals↗

Activation of 5-HT1A receptors in the medullary raphe reduces cardiovascular changes elicited by acute psychological and inflammatory stresses in rabbits.

The present strategy for the prevention of excessive sympathetic neural traffic to the heart relies on the use of beta-blockers, drugs that act at the heart end of the brain-heart axis. In the present study, we attempted to suppress cardiac sympathetic nerve activity by affecting the relevant cardiomotoneurons in the brain using the selective serotonin-1A (5-HT(1A)) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). In conscious, unrestrained rabbits, instrumented for recordings of heart rate, arterial pressure, or cardiac output, we provoked increases in cardiac sympathetic activity by psychological (loud sound, pinprick, and air jet) or inflammatory (0.5 microg/kg iv lipopolysaccharide) stresses. Pinprick and air-jet stresses elicited transient increases in heart rate (+50 +/- 7 and +38 +/- 4 beats/min, respectively) and in mean arterial pressure (+16 +/- 2 and +15 +/- 3 mmHg, respectively). Lipopolysaccharide injection caused sustained increases in heart rate (from 210 +/- 3 to 268 +/- 10 beats/min) and in arterial pressure (from 74 +/- 3 to 92 +/- 4 mmHg). Systemically administered 8-OH-DPAT (0.004-0.1 mg/kg) substantially attenuated these responses in a dose-dependent manner. Drug effects were prevented by a selective 5-HT(1A) receptor antagonist, WAY-100635 (0.1 mg/kg iv). Similarly to systemic administration, microinjection of 8-OH-DPAT (500 nl of 10 mM solution) into the medullary raphe-parapyramidal region caused antitachycardic effects during stressful stimulation and during lipopolysaccharide-elicited tachycardia. This is the first demonstration that activation of 5-HT(1A) receptors in the medullary raphe-parapyramidal area causes suppression of neurally mediated cardiovascular changes during acute psychological and immune stresses.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Inhaled nitric oxide (iNO) delivery with high-frequency jet ventilation (HFJV).

OBJECTIVE: To determine if nitric oxide (NO) therapy can be reliably administered during high-frequency jet ventilation (HFJV) using the INOvent delivery system. STUDY DESIGN: NO concentrations were measured just proximal to the endotracheal (ET) tube and at the distal tip of the ET tube during a bench evaluation. Measurements were taken over a wide range of airway pressure settings and NO concentrations with both high- and low- resistance lung models. Percent changes in set versus proximal and proximal versus distal iNO concentrations were tabulated. RESULTS: Differences between proximal and distal NO concentrations were 10% or less. In the therapeutic range of up to 20 p.p.m., differences in concentration were 1 p.p.m. or less. There was no consistent effect on NO concentration when airway resistance was increased by 500%. CONCLUSION: Nitric oxide therapy can be reliably administered during HFJV with the INOvent delivery system when NO is injected exclusively via the HFJV circuit.

Administration, Inhalation↗