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Biomedical subjects

A Berry

Publications and source records attributed to A Berry.

At least 37 records · Page 2Linked to original sources

Investigation of active structural intensity control in finite beams: theory and experiment

An investigation of structural intensity control is presented in this paper. As opposed to previous work, the instantaneous intensity is completely taken into account in the control algorithm, i.e., all the terms are considered in the real-time control process and, in particular, the evanescent waves are considered in this approach. A finite difference approach using five accelerometers is used as the sensing scheme. A feedforward filtered-X least mean square algorithm is adapted to this energy-based control problem, involving a nonpositive definite quadratic form in general. In this respect, the approach is limited to cases where the geometry is such that the intensity component will have the same sign for the control source and the primary disturbance. Results from numerical simulations are first presented to illustrate the benefit of using a cost function based on structural intensity. Experimental validation of the approach is conducted on a free-free beam covered with viscoelastic material. A comparison is made between classical acceleration control and structural intensity control and the performance of both approaches is presented. These results confirm that using intensity control allows the error sensors to be placed closer to the control source and the primary disturbance, while preserving a good control performance.

Journal Article↗

Phorbol ester induced MDR1 expression in K562 cells occurs independently of mitogen-activated protein kinase signaling pathways.

The MDR1 gene encoding the multidrug pump P-glycoprotein is transcriptionally activated in response to diverse extracellular stimuli, including the tumor promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). However, the signal transduction pathway responsible is unknown. Downstream of protein kinase C (PKC), the effects of TPA are often mediated by the Raf-1/MEK/ERK mitogen-activated protein kinase (MAPK) cascade, and Raf-1 has been implicated in MDR1 induction by serum and mitogens. Therefore, we examined the potential role of MAPK activation in TPA-mediated MDR1 induction in human leukemia K562 cells. MDR1 mRNA expression was significantly increased by TPA in the concentration range of 4 - 100 nM, with a maximal response 5 - 10 h after TPA addition. TPA-mediated MDR1 induction was inhibited by several PKC inhibitors including staurosporine, H7 and calphostin C. TPA stimulated the subcellular translocation of PKCalpha from the cytosol to the membrane and nucleus but did not affect other PKC isozymes. TPA also activated the Raf1/MEK/ERK cascade and activated another MAPK member, p38, but not JNK. In order to determine the potential role of MAPKs in MDR1 induction by TPA, specific inhibitors were utilized. The MEK inhibitor PD 098059, as well as the PKC inhibitors, completely blocked TPA-mediated ERK activation. However, under identical conditions, MDR1 induction by TPA was completely unaffected by PD 098059. Furthermore, SB 202190, which effectively inhibited TPA-mediated p38 activation, failed to inhibit TPA-induced MDR1 mRNA expression. These data demonstrate that MDR1 induction by TPA occurs via a PKC-dependent mechanism that operates independently of ERK, p38 or JNK pathways, and thus have important implications for understanding the mechanisms of MDR1 induction by extracellular stimuli.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The crystal structure of Escherichia coli class II fructose-1, 6-bisphosphate aldolase in complex with phosphoglycolohydroxamate reveals details of mechanism and specificity.

The structure of a class II fructose-1,6-bisphosphate aldolase in complex with the substrate analogue and inhibitor phosphoglycolohydroxamate (PGH) has been determined using X-ray diffraction terms to a resolution of 2.0 A (1 A=0.1 nm). The crystals are trigonal, space group P3121 with a=b=78.24 A, c=289.69 A. The asymmetric unit is a homodimer of (alpha/beta)8 barrels and the model has refined to give R-work 19.2 %, R-free (based on 5 % of the data) 23.0 %. PGH resembles the ene-diolate transition state of the physiological substrate dihydroxyacetone phosphate. It is well ordered and bound in a deep polar cavity at the C-terminal end of the (alpha/beta)8 barrel, where it chelates the catalytic zinc ion using hydroxyl and enolate oxygen atoms. Trigonal bipyramidal coordination of the zinc ion is completed by three histidine residues. The complex network of hydrogen bonds at the catalytic centre is required to organise the position of key functional groups and metal ion ligands. A well-defined monovalent cation-binding site is observed following significant re-organisation of loop structures. This assists the formation of a phosphate-binding site on one side of the barrel that tethers PGH in the catalytic site. The positions of functional groups of substrate and putative interactions with key amino acid residues are identified. Knowledge of the complex structure complements the results of spectroscopic and site-directed mutagenesis studies, and contributes to our understanding of the mechanism and substrate specificity of this family of enzymes. A reaction mechanism distinct from that proposed for other class II aldolases is discussed. The results suggest that the class II aldolases should be sub-divided into two groups on the basis of both distinct folds and mechanism.

Binding Sites↗

Conserved residues in the mechanism of the E. coli Class II FBP-aldolase.

The two classes of fructose-1,6-bisphosphate aldolase both catalyse the reversible cleavage of fructose 1,6-bisphosphate into dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. The Class I aldolases use Schiff base formation as part of their catalytic mechanism, whereas the Class II enzymes are zinc-containing metalloproteins. The mechanism of the Class II enzymes is less well understood than their Class I counterparts. We have combined sequence alignments of the Class II family of enzymes with examination of the crystal structure of the enzyme to highlight potentially important aspartate and asparagine residues in the enzyme mechanism. Asp109, Asp144, Asp288, Asp290, Asp329 and Asn286 were targeted for site-directed mutagenesis and the resulting proteins purified and characterised by steady-state kinetics using either a coupled assay system to study the overall cleavage reaction or using the hexacyanoferrate (III) oxidation of the enzyme bound intermediate carbanion to investigate partial reactions. The results showed only minor changes in the kinetic parameters for the Asp144, Asp288, Asp290 and Asp329 mutants, suggesting that these residues play only minor or indirect roles in catalysis. By contrast, mutation of Asp109 or Asn286 caused 3000-fold and 8000-fold decreases in the kcat of the reaction, respectively. Coupled with the kinetics measured for the partial reactions the results clearly demonstrate a role for Asn286 in catalysis and in binding the ketonic end of the substrate. Fourier transform infra-red spectroscopy of the wild-type and mutant enzymes has further delineated the role of Asp109 as being critically involved in the polarisation of the carbonyl group of glyceraldehyde 3-phosphate.

Escherichia coli↗

The tissue distribution of the human beta3-adrenoceptor studied using a monoclonal antibody: direct evidence of the beta3-adrenoceptor in human adipose tissue, atrium and skeletal muscle.

OBJECTIVE: To develop a monoclonal antibody that recognises an epitope of the native beta3-adrenoceptor expressed on the extracellular surface of human cells and tissues. DESIGN: A high affinity monoclonal antibody, Mab72c, was raised against the human beta3-adrenoceptor expressed on a transfected mammalian cell line. RESULTS: In CHO (Chinese hamster ovary) cells transfected with beta3-adrenoceptor cDNA, antibody labelling was found to be proportional to receptor density measured by the binding of the radiolabelled beta-adrenoceptor antagonist, [125I]-iodocyanopindolol. The use of Mab 72c has demonstrated the expression of the beta3-adrenoceptor in a variety of human tissues, including gall bladder, prostate and colon, where a mRNA signal had been detected previously. This study also provides the first direct demonstration of the expression of beta3-adrenoceptors in human skeletal muscle, atrium and adipose tissue. CONCLUSION: The development of this antibody represents an important addition to the armentarium of reagents that are available to study the localisation of beta3-adrenoceptors in human tissues.

Adipose Tissue↗

Direct measurement of mucosal pressures exerted by cuff and non-cuff portions of tracheal tubes with different cuff volumes and head and neck positions.

We measured directly mucosal pressures against the cuff and non-cuff portions of the tracheal tube in different head-neck positions and tested the reliability of calculated mucosal pressures, in vivo intracuff pressures and cuff volume as determinants of directly measured mucosal pressures. We studied 10 anaesthetized, paralysed adult patients. An 8.5-mm, high volume, low pressure PVC tracheal tube was used. Microchip sensors were attached to three cuff locations (anterior, lateral and posterior) and two non-cuff locations (anterior tip and anterior aspect of the tube, 5 cm proximal to the cuff). Directly measured mucosal pressures, in vivo intracuff pressures and calculated mucosal pressures (in vivo minus in vitro intracuff pressures) were determined after brief inflation (< 15 s) to 0, 5, 10 and 15 ml. In vivo intracuff pressures were then set at 30 mm Hg and the measurements repeated, first in the neutral position and then with the head-neck extended, flexed and rotated. Cuff mucosal pressures were highest anteriorly and lowest posteriorly. Non-cuff mucosal pressures did not vary with cuff volume and were approximately 15 mm Hg. Compared with the neutral position, in vivo intracuff pressures were higher in the rotated, extended and flexed positions. Compared with the neutral position, mucosal pressure increased on the anterior aspect of the tube in the flexed position by 22 mm Hg (P = 0.003), at the anterior tip in the extended position by 11 mm Hg (P = 0.002) and at the anterior tip (5 mm Hg, P = 0.05) and lateral aspect of the cuff (5 mm Hg, P = 0.03) in the rotated position. In vivo intracuff pressures and calculated mucosal pressures were moderate predictors of measured mucosal pressures; cuff volume was a poor predictor. We conclude that tracheal mucosal pressures were highest anteriorly, that non-cuff portions of the tube exerted substantial mucosal pressures and that the rotated position caused a greater increase in tracheal mucosal pressure than the extended or flexed position. Indirect methods of measuring mucosal pressure were of moderate predictive value.

Adult↗

Pulmonary airway resistance with the endotracheal tube versus laryngeal mask airway in paralyzed anesthetized adult patients.

BACKGROUND: The hypothesis that airway resistance is less with the laryngeal mask airway than with the endotracheal tube was tested. METHODS: Thirty-six paralyzed, anesthetized adult patients with no respiratory disease (American Society of Anesthesiologists physical status 1-3; age, 18-80 yr) were randomly allocated (9 men, 9 women in each group) to receive either a size-4 laryngeal mask airway or an endotracheal tube (men, 9-mm ID; women, 8-mm ID). A pulmonary monitor with flow transducer and esophageal balloon was used to measure peak airway pressure and mean airway resistance (device resistance plus pulmonary airway resistance) at three different tidal volumes (5, 10, and 15 ml/kg). Device resistance was measured in vitro with the distal end of the endotracheal tube or laryngeal mask airway open to the atmosphere and using the same ventilator settings. Pulmonary airway resistance was derived by subtracting the mean device resistance from the mean airway resistance. RESULTS: Peak airway pressure, mean airway resistance, device resistance, and pulmonary airway resistance were greater for the endotracheal tube (all P < 0.0001). CONCLUSIONS: The laryngeal mask airway triggers less bronchoconstriction than does the endotracheal tube in paralyzed anesthetized adult patients. This may have implications for maintaining intraoperative pulmonary function and reducing the risk for atelectasis and pulmonary infection.

Adolescent↗

The dhnA gene of Escherichia coli encodes a class I fructose bisphosphate aldolase.

The gene encoding the Escherichia coli Class I fructose-1, 6-bisphosphate aldolase (FBP aldolase) has been cloned and the protein overproduced in high amounts. This gene sequence has previously been identified as encoding an E. coli dehydrin in the GenBanktrade mark database [gene dhnA; entry code U73760; Close and Choi (1996) Submission to GenBanktrade mark]. However, the purified protein overproduced from the dhnA gene shares all its properties with those known for the E. coli Class I FBP aldolase. The protein is an 8-10-mer with a native molecular mass of approx. 340 kDa, each subunit consisting of 349 amino acids. The Class I enzyme shows low sequence identity with other known FBP aldolases, both Class I and Class II (in the order of 20%), which may be reflected by some novel properties of this FBP aldolase. The active-site peptide has been isolated and the Schiff-base-forming lysine residue (Lys236) has been identified by a combination of site-directed mutagenesis, kinetics and electrospray-ionization MS. A second lysine residue (Lys238) has been implicated in substrate binding. The cloning of this gene and the high levels of overexpression obtained will facilitate future structure-function studies.

Amino Acid Sequence↗

The cuffed oropharyngeal airway for spontaneous ventilation anaesthesia. Clinical appraisal in 100 patients.

The cuffed oropharyngeal airway is a modified Guedel airway with a distal inflatable cuff and a proximal connector for attachment to an anaesthesia circuit. The purpose of this study was to evaluate this device for spontaneous ventilation anaesthesia in 100 adult patients. Anaesthesia was induced with fentanyl and propofol and maintained with nitrous oxide and isoflurane in oxygen. The device was inserted when anaesthetic depth was judged to be adequate. Adverse airway events and interventions (i.e., manoeuvres performed in order to provide a clear airway) were analysed from video recordings and detailed notes. The position of the device was assessed fibreoptically during spontaneous ventilation. The device provided a clear airway in 98% of patients during manually assisted ventilation, in 100% during spontaneous ventilation and in 100% during emergence. However, 91% of patients required at least one airway intervention at some time. Jaw life was required for 33% of the time during manually assisted ventilation and 21% of the time during spontaneous ventilation. Most interventions occurred during the first 3 min of either manually assisted or spontaneous ventilation. The incidence of adverse airway events during manually assisted ventilation was 8%, during spontaneous ventilation was 5% and during emergence was 5%. Oxygen saturation briefly fell to between 87 and 89% on six occasions. On fibreoptic assessment, the vocal cords were visible in 29% of patients on fibreoptic assessment and the epiglottis was visible in 90%. Mild sore throat occurred in 4% of patient. We conclude that the cuffed oropharyngeal airway is suitable for spontaneous ventilation anaesthesia and has a low complication rate but that most patients require one or more interventions to provide a clear airway.

Adolescent↗

A randomized controlled trial comparing the cuffed oropharyngeal airway and the laryngeal mask airway in spontaneously breathing anesthetized adults.

BACKGROUND: The cuffed oropharyngeal airway (COPA), a modified Guedel airway, was compared with the laryngeal mask airway (LMA) during spontaneous breathing anesthesia. Specifically examined were ease of use, physiologic tolerance, and the frequency of problems. METHODS: Adult patients consented to random (2:1) assignment to either COPA (n = 302) or LMA (n = 151) for airway management during anesthesia with propofol, nitrous oxide, and oxygen. RESULTS: Ease of insertion was similar, but the first-time successful insertion rate was higher with the LMA (COPA, 81% compared with LMA, 89%; P = 0.05). More brief manipulations (head tilt, chin lift, jaw thrust) were reported in the COPA group (average total number of manipulations: COPA, 1.1 +/- 1.6 compared with LMA, 0.1 +/- 0.2; P < 0.001). Continuous airway support was used more frequently in the COPA group (COPA, 30% compared with LMA, 0%; P < 0.0005). The incidences of aspiration, regurgitation, laryngospasm, wheezing, succinylcholine administration, oxygen saturation (SpO2) < 92%, failed use, and minor intraoperative problems were similar. When the airways were removed, blood was detected on the COPA less frequently than on the LMA (COPA, 5.8% compared with LMA, 15.3%; P = 0.001). The incidence of early and late sore throat was greater with the LMA (early: COPA, 4.7% compared with LMA, 21.9% [P = 0.001]; late: COPA, 8.4% compared with LMA, 16.1%; P = 0.01). The LMA did better than the COPA when anesthetists analyzed the technical aspects of the two devices. CONCLUSIONS: Although the COPA and LMA are equivalent devices in terms of physiologic alterations and overall clinical problems associated with their use, the LMA was associated with a higher first-time insertion rate and fewer manipulations, suggesting that it is easier to use. The COPA was associated with less blood on the device and fewer sore throats, suggesting it may cause less pharyngeal trauma. Ultimately, both devices were similar in establishing a safe and effective airway for spontaneously breathing anesthetized adults.

Adult↗

Changes in membrane fluidity induced by binding of immunoglobulins to sperm Fc receptors.

Receptors for Fc region of immunoglobulins (FcR) are reported to be present on spermatozoa, and also detected in seminal plasma, however their function is still not known. Since various changes in sperm membrane architecture during maturation and passage through female genital tract are reported, experiments were conducted to study the membrane fluidity changes in sperm subsequent to ligation of surface FcR with immunoglobulin and its derivatives. This paper reports that interaction with IgG restricts rotational mobility of cell surface proteins and membrane lipids as studied by EPR spectroscopy using spin probes. Decrease in fluidity was much more pronounced in the presence of aggregated IgG due to crosslinking of sperm FcR by aggregated IgG.

Animals↗