The importance of recognising Streptococcus milleri as a cause of orbital cellulitis.
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Biomedical subjects
Publications and source records attributed to P Leong.
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A computational model of auditory localization resulting in performance similar to humans is reported. The model incorporates both the monaural and binaural cues available to a human for sound localization. Essential elements used in the simulation of the processes of auditory cue generation and encoding by the nervous system include measured head-related transfer functions (HRTFs), minimum audible field (MAF), and the Patterson-Holdsworth cochlear model. A two-layer feed-forward back-propagation artificial neural network (ANN) was trained to transform the localization cues to a two-dimensional map that gives the direction of the sound source. The model results were compared with (i) the localization performance of the human listener who provided the HRTFs for the model and (ii) the localization performance of a group of 19 other human listeners. The localization accuracy and front-back confusion error rates exhibited by the model were similar to both the single listener and the group results. This suggests that the simulation of the cue generation and extraction processes as well as the model parameters were reasonable approximations to the overall biological processes. The amplitude resolution of the monaural spectral cues was varied and the influence on the model's performance was determined. The model with 128 cochlear channels required an amplitude resolution of approximately 20 discrete levels for encoding the spectral cue to deliver similar localization performance to the group of human listeners.
Globus pharyngeus as a possible presenting symptom of a pharyngeal or upper oesophageal neoplasm is the main reason why barium swallows are requested, although it is essentially a benign disorder that in many cases requires reassurance only. We therefore retrospectively reviewed all barium swallows done in our department for globus pharyngeus during a one-year period to assess their value in the investigation of this condition. Ninety-two patients were identified. All had a normal ENT examination, and symptoms suggestive of acid reflux was the most common associated complaint, at 11 per cent. Acid reflux (18.5 per cent) and hiatus hernia (12 per cent) were the commonest findings of a barium swallow. Three cases of suspicious findings on barium swallow had a normal examination under general anaesthetic. Statistical analysis showed no significant relationship between the symptoms of globus and the barium swallow results. A barium swallow does not seem to add any further useful information to the investigation of globus pharyngeus. But most globus patients will continue to undergo a barium swallow, because although alternative investigations have been extensively assessed, it is still not clear which is the most appropriate mode of investigation for this condition.
Wnt and its intracellular effector beta-catenin regulate developmental and oncogenic processes. Using expression cloning to identify novel components of the Wnt pathway, we isolated casein kinase Iepsilon (CKIepsilon). CKIepsilon mimicked Wnt in inducing a secondary axis in Xenopus, stabilizing beta-catenin, and stimulating gene transcription in cells. Inhibition of endogenous CKIepsilon by kinase-defective CKIepsilon or CKIepsilon antisense-oligonucleotides attenuated Wnt signaling. CKIepsilon was in a complex with axin and other downstream components of the Wnt pathway, including Dishevelled. CKIepsilon appears to be a positive regulator of the pathway and a link between upstream signals and the complexes that regulate beta-catenin.
Exclusive aplasia of maxillary permanent canines is extremely rare. There are only a few cases of this condition reported in the literature. This paper reports a case of bilateral congenitally missing canines in a healthy 11 year old male of Chinese origin. The article discusses problems in diagnosis and presents options in the management of such a case. The management option selected was the most appropriate for the family at the time of presentation.
Excitation-contraction coupling in skeletal muscle is a result of the interaction between the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor or RyR1) and the skeletal muscle L-type Ca2+ channel (dihydropyridine receptor or DHPR). Interactions between RyR1 and DHPR are critical for the depolarization-induced activation of Ca2+ release from the sarcoplasmic reticulum, enhancement of DHPR Ca2+ channel activity, and repolarization-induced inactivation of RyR1. The DHPR III-IV loop was fused to glutathione S-transferase (GST) or His-peptide and used as a protein affinity column for 35S-labeled, in vitro translated fragments from the N-terminal three-fourths of RyR1. RyR1 residues Leu922-Asp1112 bound specifically to the DHPR III-IV loop column, but the corresponding fragment from the cardiac ryanodine receptor (RyR2) did not. Construction of chimeras between RyR1 and RyR2 showed that amino acids Lys954-Asp1112 retained full binding activity, whereas Leu922-Phe1075 had no binding activity. The RyR1 sequence Arg1076-Asp1112, previously shown to interact with the DHPR II-III loop (Leong, P., and MacLennan, D., H. (1998) J. Biol. Chem. 273, 7791-7794), bound to DHPR III-IV loop columns, but with only half the efficiency of binding of the longer RyR1 sequence, Lys954-Asp1112. These data suggest that the site of DHPR III-IV loop interaction contains elements from both the Lys954-Phe1075 and Arg1076-Asp1112 fragments. The presence of 4 +/- 0.4 microM GST-DHPR II-III or 5 +/- 0.1 microM His-peptide-DHPR III-IV was required for half-maximal co-purification of 35S-labeled RyR1 Leu922-Asp1112 on glutathione-Sepharose or Ni2+-nitrilotriacetic acid. Dose-dependent inhibition of 35S-labeled RyR1 Leu922-Asp1112 binding to GST-DHPR II-III and GST-DHPR III-IV by His10-DHPR II-III and His-peptide-DHPR III-IV was observed. These studies indicate that the DHPR II-III and III-IV loops bind to contiguous and possibly overlapping sites on RyR1 between Lys 954 and Asp1112.
A systematic analysis of the localization of objects in extra-personal space requires a three-dimensional method of documenting location. In auditory localization studies the location of a sound source is often reduced to a directional vector with constant magnitude with respect to the observer, data being plotted on a unit sphere with the observer at the origin. This is an attractive form of data representation as the relevant spherical statistical and graphical methods are well described. In this paper we collect together a set of spherical plotting and statistical procedures to visualize and summarize these data. We describe methods for visualizing auditory localization data without assuming that the principal components of the data are aligned with the coordinate system. As a means of comparing experimental techniques and having a common set of data for the verification of spherical statistics, the software (implemented in MATLAB) and database described in this paper have been placed in the public domain. Although originally intended for the visualization and summarization of auditory psychophysical data, these routines are sufficiently general to be applied in other situations involving spherical data.
Interactions between the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor or RyR1) and the loop linking domains II and III (II-III loop) of the skeletal muscle L-type Ca2+ channel (dihydropyridine receptor or DHPR) are critical for excitation-contraction coupling in skeletal muscle. The DHPR II-III loop was fused to glutathione S-transferase- or His-peptide and used as a protein affinity column for 35S-labeled in vitro translated fragments from the N-terminal three-fourths of RyR1. RyR1 residues Leu922-Asp1112 bound specifically to the DHPR II-III loop column, but the corresponding fragment from the cardiac ryanodine receptor (RyR2) did not. The use of chimeras between RyR1 and RyR2 localized the interaction to 37 amino acids, Arg1076-Asp1112, in RyR1. The RyR1 922-1112 fragment did not bind to the cardiac DHPR II-III loop but did bind to the skeletal muscle Na+ channel II-III loop. The skeletal DHPR II-III loop double mutant K677E/K682E lost most of its capacity to interact with RyR1, suggesting that two positively charged residues are important in the interaction between RyR and DHPR.
The operation of Laser Assisted Uvulopalatoplasty (LAUP) as described by Kamami is now becoming more commonly used in the treatment of snoring and obstructive sleep apnoea. The authors have treated 95 snoring patients, varying the lengths of the soft palate incisions and percentage of uvula excised. All operations were carried out under general anaesthesia using a CO2 laser. Pilot studies showed incisions that are 25% of the distance between the free edge of the soft palate to the hard palate junction and excision of 50% of the uvula give good results with minimal complications. A further study using these parameters was conducted and postoperative evaluation including polysomnography confirmed this procedure to be effective in reducing snoring levels both subjectively and objectively.
Evidence for functional interactions between the Ca2+ release channel in the skeletal muscle sarcoplasmic reticulum (the ryanodine receptor) and the L-type Ca2+ channel in the sarcolemma (the dihydropyridine receptor), leading to excitation-contraction coupling, is reviewed and experimental systems used to identify candidate sites of interaction are outlined.
We report transient expression of a full-length cDNA encoding the Ca2+ release channel of rabbit skeletal muscle sarcoplasmic reticulum (ryanodine receptor) in HEK-293 cells. The single-channel properties of the 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate-solubilized and sucrose gradient-purified recombinant Ca2+ release channels were investigated by using single-channel recordings in planar lipid bilayers. The recombinant Ca2+ release channel exhibited a K+ conductance of 780 pS when symmetrical 250 mM KCl was used as the conducting ion and a Ca2+ conductance of 116 pS in 50 mM luminal Ca2+. Opening events of the recombinant channels were brief, with an open time constant of approximately 0.22 ms. The recombinant Ca2+ release channel was more permeable to Ca2+ than to K+, with a pCa2+/pK+ ratio of 6.8. The response of the recombinant Ca2+ release channel to various concentrations of Ca2+ was biphasic, with the channel being activated by micromolar Ca2+ and inhibited by millimolar Ca2+. The recombinant channels were activated by ATP and caffeine, inhibited by Mg2+ and ruthenium red, and modified by ryanodine. Most recombinant channels were asymmetrically blocked, conducting current unidirectionally from the luminal to the cytoplasmic side of the channel. These data demonstrate that the properties of recombinant Ca2+ release channel expressed in HEK-293 cells are very similar, if not identical, to those of the native channel.
Measurement of localization performance will reflect errors that relate to the sensory processing of the cues to sound location and the errors associated with the method by which the subject indicates the perceived location. This study has measured the ability of human subjects to localize a short noise burst presented in the free field with the subject indicating the perceived location by pointing their nose towards the source. Subjects were first trained using a closed loop training paradigm which involved instantaneous feedback as to the accuracy of head pointing which resulted in the reduction of residual localization errors and a rapid acquisition of the task by the subjects. Once trained, 19 subjects localized between 4 and 6 blocks of 76 target locations. The data were pooled and the distribution of errors associated with each target location was examined using spherical methods. Errors in the localization estimates for about one third of the locations were rotationally symmetrical about their mean but the remaining locations were best described by an elliptical distribution (Kent distributed). For about one half of the latter locations the orientations of the directions of the greatest variance of the distributions were not aligned with the azimuth and elevation coordinates used for describing the spatial location of the targets. The accuracy (systematic errors) and the distribution of the errors (variance) in localization for our population of subjects were also examined for each test location. The size of the data set and the methods of analysis provide very reliable measures of important baseline parameters of human auditory localization.
Anaphylaxis is one of the most catastrophic, although infrequent, potential side effects of chemotherapy. We report the case of a patient who developed respiratory distress and who could not be resuscitated after his sixth cycle of cisplatin and paclitaxel for squamous carcinoma of the lung and false vocal cord. Autopsy confirmed anaphylaxis as the cause of death. The major hypersensitivity profiles of these two agents, as described in case reports and in the medical literature, were reviewed to determine which of the two therapies was the underlying cause of the patient's reaction. Anaphylaxis has been reported with both cisplatin and paclitaxel. Cisplatin is argued to be the most likely etiology for anaphylaxis in this case.
Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.
UDP-GlcNAc:alpha-6-D-mannoside [GlcNAc to Man alpha 1-6] beta-1,2-N-acetylglucosaminyltransferase II (GlcNAc-T II, EC 2.4.1.143) is a Golgi enzyme catalyzing an essential step in the conversion of oligomannose to complex N-glycans. A 1.2-kb probe from a rat liver cDNA encoding GlcNAc-T II was used to screen a human genomic DNA library in lambda EMBL3. Southern analysis of restriction endonuclease digests of positive phage clones identified two hybridizing fragments (3.0 and 3.5 kb) which were subcloned into pBlueScript. The inserts of the resulting plasmids (pHG30 and pHG36) are over-lapping clones containing 5.5 kb of genomic DNA. The pHG30 insert (3.0 kb) contains a 1341-bp open reading frame encoding a 447-amino-acid protein, 250 bp of G + C-rich 5'-upstream sequence and 1.4 kb of 3'-downstream sequence. The pHG36 insert (3.5 kb) contains 2.75 kb of 5'-upstream sequence and 750 bp of the 5'-end of the open reading frame. The protein sequence showed the domain structure typical of all previously cloned glycosyltransferases, i.e. a short 9-residue putative cytoplasmic N-terminal domain, a 20-residue hydrophobic non-cleavable putative signal-anchor domain and a 418-residue C-terminal catalytic domain. Northern analysis of human tissues showed a major message at 3 kb and minor signals at 2 and 4.5 kb. There is no sequence similarity to any previously cloned glycosyltransferases including human UDP-GlcNAc:alpha-3-D-mannoside [GlcNAc to Man alpha 1-3] beta-1,2-N-acetylglucosaminyltransferase I (GlcNAc-T I) which has 445 amino acids with a 418-residue C-terminal catalytic domain. The human GlcNAc-T I and II genes (MGAT1 and MGAT2) map to chromosome bands 5q35 and 14q21, respectively, by fluorescence in situ hybridization. The entire coding regions of human GlcNAc-T I and II are each on a single exon. There is 92% identity between the amino acid sequences of the catalytic domains of human and rat GlcNAc-T II. Southern analysis of restriction enzyme digests of human genomic DNA indicates that there is only a single copy of the MGAT2 gene. The full-length coding region of GlcNAc-T II has been expressed in the baculovirus/Sf9 insect cell system, the recombinant enzyme has been purified to near homogeneity with a specific activity of about 20 mumol.min-1.mg-1 and the product synthesized by the recombinant enzyme has been identified by high-resolution 1H-NMR spectroscopy and mass spectrometry.
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The relative antiandrogen-induced prostate involution activity of the newly synthesized hydroxyflutamide pro-drug was compared with that of flutamide in 25 Beagles. Secondary antiandrogen activity of both drugs on the testes and mammary tissue was investigated. Daily oral administration of both compounds at 2 dosages (ie, 2.5 and 5.0 mg/kg of body weight) during a 7-week period was monitored by transabdominal ultrasonography of the prostate twice a week. Cross-sectional area estimates of the prostate gland calculated from oblique dorsoventral, and transverse sonographic measurements were diminished significantly in some of the treated dogs as early as day 14 of drug administration. All treated dogs had significant differences in reduction by day 47. Involution was related directly to dose (P less than 0.05), but no difference was observed between test compounds. Differences in secondary antiandrogen activity were not remarkable. Flutamide was not found to have any activity advantage in vivo over hydroxyflutamide. It was concluded that ultrasonography can be a highly effective means of monitoring prostate size, and of monitoring drug-induced involution over time.
The DNA base sequence changes induced by mutagenesis with ultraviolet light have been determined in a gene on a chromosome of cultured Chinese hamster ovary (CHO) cells. The gene was the Escherichia coli gpt gene, of which a single copy was stably incorporated and expressed in the CHO cell genome. The cells were irradiated with ultraviolet light and gpt- colonies were selected by resistance to 6-thioguanine. The gpt gene was amplified from chromosomal DNA by use of the polymerase chain reaction (PCR), and the amplified DNA sequenced directly by the dideoxy method. Of the 58 sequenced mutants of independent origin 53 were base change mutations. Forty-one base substitutions were single base changes, ten had two adjacent (or tandem) base changes, and one had two base changes separated by a single base-pair. Only one mutant had a multiple base change mutation with two or more well separated base changes. In contrast much higher levels of such mutations were reported in ultraviolet mutagenesis of genes on a shuttle vector in primate cells. Two deletions of a single base-pair were observed and three deletions ranging from 6 to 37 base-pairs. The mutation spectrum in the gpt gene had similarities to the ultraviolet mutation spectra for several genes in prokaryotes, which suggests similarities in mutational mechanisms in prokaryotes and eukaryotes.