Antiplatelet drugs in elective ENT surgery.

INTRODUCTION: Oral antiplatelet drugs are increasingly being encountered in patients scheduled for elective ENT surgery. Their pre-operative cessation can have potentially serious complications in some patients, particularly those with intracoronary stents. METHODS: In order to gain an impression of current peri-operative management of patients taking antiplatelet drugs, an online survey was distributed to the Expert Panel of ENT UK, the British Association of Otolaryngologists Head and Neck Surgeons, between 13 January and 15 February 2011. RESULTS: Three hundred and three members were contacted. The response rate was 55 per cent (167 replies); 78 per cent of respondents were consultants. Results are presented in the main text. CONCLUSION AND RECOMMENDATIONS: Patients can be categorised as high or low risk, depending on their indication for taking antiplatelet drugs. Recommendations taken from the literature are given on how best to manage these two groups.

Experimental evidence for two-step nucleation in colloidal crystallization.

We investigated the freezing of colloidal spheres in two dimensions with single-particle resolution. Using micron-size, charge-stabilized polystyrene spheres with a temperature-dependent depletion attraction induced by surfactant micelles, we supercooled an initially amorphous (gaslike) system. Particle motions were monitored as crystallization proceeded. At low concentrations, freezing occurred in a single step in a manner consistent with classical nucleation theory. In other samples two-step nucleation was found, in which amorphous clusters grew to approximately 30 particles, then rapidly crystallized. Measured free energies show the role of metastable gas-liquid coexistence, which also enhanced the rate of nucleation following deeper quenches.

Spontaneous descending retropharyngeal abscess.

Retropharyngeal abscesses are rare but can occur spontaneously in adults and are potentially life threatening. Such a diagnosis should be considered so that aggressive treatment can be initiated as soon as possible to avoid life threatening mediastinal complications.

Imaging the sublimation dynamics of colloidal crystallites.

We studied the kinetics of sublimating crystals with single-particle resolution by experiments with colloidal spheres and by computer simulations. A short-range attraction between spheres led to crystallites one to three layers thick. The spheres were tracked with optical microscopy while the attraction was reduced and the crystals sublimated. Large crystallites sublimated by escape of particles from the perimeter. The rate of shrinkage was greatly enhanced, however, when the size decreased to less than 20 to 50 particles, depending on the location in the phase diagram. At this size, the crystallites transformed into a dense amorphous structure, which rapidly vaporized. The enhancement of kinetics by metastable or unstable phases may play a major role in the melting, freezing, and annealing of crystals.

Litigation in otolaryngology - trends and recommendations.

BACKGROUND: Seeking to identify where litigious claims against otolaryngologists are targeted (i.e. areas of highest risk) within the NHS and private sector would have positive implications in risk management and limiting the amount of litigation against otolaryngologists. METHOD: The National Health Service Litigation Authority (NHSLA) and Medical Defence Union (MDU) were contacted and anonymous data obtained on claims within ENT. RESULTS: 887 claims were notified - 457 NHSLA and 430 MDU. The commonest claim in both groups was failure or delay in diagnosis (12 per cent NHSLA, 23 per cent MDU). The other commonest claims were all related to complications (nerve damage 10 per cent, deafness 8 per cent and dental damage 5 per cent). Dissatisfaction with results was 8 per cent total and, within the private sector, was almost exclusively in rhinology. CONCLUSIONS: This study once again emphasizes the need for thorough clinical assessment, record keeping and good communication with patients. Recognising these areas of highest risk may limit future claims.

A new combined approach to the nasopharynx in head and neck plastic surgery: technique and application.

This article presents a combined approach to the nasopharynx, which has not been previously described. The technique is applicable to cases of recurrent nasopharyngeal carcinoma which exhibit lateral extension. We describe the technique and report a case in which it was used. A review is presented of all other techniques currently described. The authors would not advocate the use of a combined approach to the nasopharynx in all cases, but there certainly appears to be a place for this technique.

On the nature of visible chromosomal gaps and breaks.

From the earliest days of chromosomal aberration studies, the distinction, nature and origin of light-microscope observed "gaps" and "breaks" have been topics for debate and controversy. In this paper we survey, briefly, the various ideas that have appeared in the very extensive literature, and attempt to evaluate them in the light of our current understanding of chromosome structure and aberration formation. Attention is drawn to the problems of interpretation caused by G2/S cell imprecision.

Chromosomal aberrations: formation, identification and distribution.

Chromosomal aberrations (CA) are the microscopically visible part of a wide spectrum of DNA changes generated by different repair mechanisms of DNA double strand breaks (DSB). The method of fluorescence in situ hybridisation (FISH) has uncovered unexpected complexities of CA and this will lead to changes in our thinking about the origin of CA. The inter- and intrachromosomal distribution of breakpoints is generally not random. CA breakpoints occur preferentially in active chromatin. Deviations from expected interchromosomal distributions of breakpoints may result from the arrangement of chromosomes in the interphase nucleus and/or from different sensitivities of chromosomes with respect to the formation of CA. Telomeres and interstitial telomere repeat like sequences play an important role in the formation of CA. Subtelomeric regions are hot spots for the formation of symmetrical exchanges between homologous chromatids and cryptic aberrations in these regions are associated with human congenital abnormalities.

Modulation of the G protein regulator phosducin by Ca2+/calmodulin-dependent protein kinase II phosphorylation and 14-3-3 protein binding.

Phototransduction is a canonical G protein-mediated cascade of retinal photoreceptor cells that transforms photons into neural responses. Phosducin (Pd) is a Gbetagamma-binding protein that is highly expressed in photoreceptors. Pd is phosphorylated in dark-adapted retina and is dephosphorylated in response to light. Dephosphorylated Pd binds Gbetagamma with high affinity and inhibits the interaction of Gbetagamma with Galpha or other effectors, whereas phosphorylated Pd does not. These results have led to the hypothesis that Pd down-regulates the light response. Consequently, it is important to understand the mechanisms of regulation of Pd phosphorylation. We have previously shown that phosphorylation of Pd by cAMP-dependent protein kinase moderately inhibits its association with Gbetagamma. In this study, we report that Pd was rapidly phosphorylated by Ca(2+)/calmodulin-dependent kinase II, resulting in 100-fold greater inhibition of Gbetagamma binding than cAMP-dependent protein kinase phosphorylation. Furthermore, Pd phosphorylation by Ca(2+)/calmodulin-dependent kinase II at Ser-54 and Ser-73 led to binding of the phosphoserine-binding protein 14-3-3. Importantly, in vivo decreases in Ca(2+) concentration blocked the interaction of Pd with 14-3-3, indicating that Ca(2+) controls the phosphorylation state of Ser-54 and Ser-73 in vivo. These results are consistent with a role for Pd in Ca(2+)-dependent light adaptation processes in photoreceptor cells and also suggest other possible physiological functions.

Collaborative exercise on the use of FISH chromosome painting for retrospective biodosimetry of Mayak nuclear-industrial personnel.

PURPOSE: To investigate within the framework of a multilaboratory study the suitability of FISH chromosome painting to measure so-called stable translocations in peripheral lymphocytes of Mayak nuclear-industrial workers (from the Southern Urals) and their use for retrospective biodosimetry. MATERIALS AND METHODS: Chromosime analyses were carried out from 69 workers who had received protracted occupational radiation exposures (0.012-6.065 Gy) up to approximately 40 years before blood sampling. Twenty-one unexposed people living in the same area were controls. A multicolour FISH-painting protocol with the target chromosomes 1, 4 and 8 simultaneously with a pancentromeric probe was used to score potentially transmissible chromosome-type aberrations (reciprocal translocations 2B and related 'one-way' patterns I-III according to the S&S classification). RESULTS: Individual biodosimetry estimates were obtained in terms of these potentially long-term surviving aberration types based on the linear component of a low dose-rate gamma-ray calibration curve produced using identical staining and scoring protocols. For comparison, the workers personal and total background doses were converted to red bone marrow doses. The estimated doses were mainly lower than would be predicted by the calibration curve, particularly at accumulated higher dose levels. CONCLUSIONS: Owing to the limited life-time of circulating T-lymphocytes, the long-term persistence of translocations in vivo requires the assumption of a clonal repopulation of these naturally senescing cells from the haemopoietic stem cell compartments. Obviously such a replacement cannot be fully achieved, leading to a temporal decline even of the yield of transmissible aberrations types. Assuming further a highly selective capacity of stem cells against any type of chromosomal damage and the fact that one must rely on partial genome findings, the potential of FISH chromosome painting for retrospective dose reconstruction is probably limited to a decade or so after high-level protracted radiation exposure.

Functional roles of the two domains of phosducin and phosducin-like protein.

Phosducin and phosducin-like protein regulate G protein signaling pathways by binding the betagamma subunit complex (Gbetagamma) and blocking Gbetagamma association with Galpha subunits, effector enzymes, or membranes. Both proteins are composed of two structurally independent domains, each constituting approximately half of the molecule. We investigated the functional roles of the two domains of phosducin and phosducin-like protein in binding retinal G(t)betagamma. Kinetic measurements using surface plasmon resonance showed that: 1) phosducin bound G(t)betagamma with a 2. 5-fold greater affinity than phosducin-like protein; 2) phosphorylation of phosducin decreased its affinity by 3-fold, principally as a result of a decrease in k(1); and 3) most of the free energy of binding comes from the N-terminal domain with a lesser contribution from the C-terminal domain. In assays measuring the association of G(t)betagamma with G(t)alpha and light-activated rhodopsin, both N-terminal domains inhibited binding while neither of the C-terminal domains had any effect. In assays measuring membrane binding of G(t)betagamma, both the N- and C-terminal domains inhibited membrane association, but much less effectively than the full-length proteins. This inhibition could only be described by models that included a change in G(t)betagamma to a conformation that did not bind the membrane. These models yielded a free energy change of +1.5 +/- 0.25 kcal/mol for the transition from the G(t)alpha-binding to the Pd-binding conformation of G(t)betagamma.

Underprediction of visibly complex chromosome aberrations by a recombinational-repair ('one-hit') model.

PURPOSE: Published low-LET FISH data were used to test two models of chromosome aberration production based on breakage-and-reunion or recombinational repair. MATERIALS AND METHODS: Randomness of DNA double strand break induction and misrejoining is analyzed comprehensively and adopted as a working hypothesis. Proximity effects are approximated by using interaction sites. Model results are calculated using CAS (chromosome aberration simulator) Monte Carlo computer software with two adjustable parameters. CAS can emulate the specifics of any experimental painting protocol, allowing very detailed tests of the models. RESULTS: To reasonable approximation, breakage-and-reunion model predictions are consistent with low-LET FISH results, including two large, elaborate, one-paint data sets. An explicitly specified version of the recombinational-repair model severely underpredicts the frequency of the visibly complex aberration patterns most commonly observed with one-paint FISH, and is inconsistent with some observed multi-paint patterns. When high-dose effects (distortion and saturation) are taken into account quantitatively, a dose-response relation for apparently simple interchanges slightly favours the breakage-and-reunion model over the recombinational-repair model, despite being approximately linear over the dose range 2-6 Gy. CONCLUSIONS: The random breakage-and-reunion model gives comprehensive baseline predictions that are sufficiently accurate for the organization of experimental results. The data speak against complex aberrations being formed by the random recombinational repair pathway discussed here.

A molecular mechanism for the phosphorylation-dependent regulation of heterotrimeric G proteins by phosducin.

Visual signal transduction is a nearly noise-free process that is exquisitely well regulated over a wide dynamic range of light intensity. A key component in dark/light adaptation is phosducin, a phosphorylatable protein that modulates the amount of transducin heterotrimer (Gt alpha beta gamma) available through sequestration of the beta gamma subunits (Gt beta gamma). The structure of the phosphophosducin/Gt beta gamma complex combined with mutational and biophysical analysis provides a stereochemical mechanism for the regulation of the phosducin-Gt beta gamma interaction. Phosphorylation of serine 73 causes an order-to-disorder transition of a 20-residue stretch, including the phosphorylation site, by disrupting a helix-capping motif. This transition disrupts phosducin's interface with Gt beta gamma, leading to the release of unencumbered Gt beta gamma, which reassociates with the membrane and Gt alpha to form a signaling-competent Gt alpha beta gamma heterotrimer.

X-ray-induced simple, pseudosimple and complex exchanges involving two distinctly painted chromosomes.

PURPOSE: To detect simple, pseudosimple and complex chromosome exchanges in X-ray-induced aberrations involving two distinctly painted chromosomes. Each visibly complex two-paint exchange was analysed to determine the number of breaks and chromosomes necessary to derive the pattern. In addition, the number of associated paint junctions was scored to assess the frequency of non-reciprocal exchanges. MATERIALS AND METHODS: Metaphase spreads were prepared from a human primary fibroblast cell line irradiated with 2, 4 and 6 Gy 250kV X-rays. FISH-painting was performed with distinctly labelled probes for chromosomes 1 and 2, and a pancentromeric probe. RESULTS: From a total of 78 two-paint exchanges observed, 35 were apparently simple, with no additional counterstain chromatin, and 43 were visibly complex with two-colour painting, of which 23 contained at least one pseudosimple exchange. A detailed analysis of the number of two-paint colour junctions showed that at least 50% of the visibly complex exchange patterns involved non-reciprocal exchanges. The simple and complex exchange dose-response curves were considered to be linear and curvilinear respectively. CONCLUSION: The frequency of non-reciprocal rejoining events within complex exchanges is consistent with an interaction model based on the free exchange of multiple break-ends. In addition, the simple and complex exchanges have distinct dose-response curves, in agreement with previous data for single-painted exchanges corrected for pseudosimples.

Interphase chromosome domain re-organization following irradiation.

PURPOSE: To investigate chromosome domain movements during interphase in stationary cell nuclei. MATERIALS AND METHODS: Contact-inhibited primary human fibroblasts were irradiated with 4.0 Gy X-rays, BrdU was added, and air-dried cell preparations made at intervals up to 48 h. Chromosome 4 domain signals (1, 2 and >2) in BrdU negative nuclei (almost exclusively G0/G1) were counted using FISH. A similar experiment was performed using unstimulated human lymphocytes. RESULTS: A very significant rise in nuclei with >2 signals was found within 1 h after radiation. The frequencies observed were in very good agreement with those expected for simple and complex interchanges involving chromosome 4, scored at metaphase in these materials. CONCLUSIONS: The observations constitute evidence for significant domain movement and re-organization within a short time of radiation exposure in G0/G1 interphase nuclei, presumably induced by the formation of inter-domain exchanges. Such re-organization must be a very complicated and delicate topological problem for relaxed chromatin, and must have an important bearing on the interpretation of mechanistic premature chromosome condensation experiments performed whilst it is in operation.

Clustering of radiation-produced breaks along chromosomes: modelling the effects on chromosome aberrations.

PURPOSE: For high-LET radiations, and perhaps even for hard X-rays, DNA double-strand breaks (dsb) are clustered nonrandomly along chromosomes; disproportionately, many inter-dsb segments are less than a few Mbp (10(6) base pairs). The implications of such dsb clustering for chromosome aberrations are analysed. METHODS: Chromosome segments between different dsb within one dsb cluster are assumed too small to detect in the aberration assay. Enumeration or Monte-Carlo computer simulations are used to compute the relative frequencies of many observable aberration patterns: apparently simple or visibly complex. The theoretical predictions are compared with X-ray data for human fibroblasts, involving painted chromosomes 1, 2, 4, 5, 7 or 13. RESULTS AND CONCLUSIONS: Surprisingly, cryptic dsb multiplicity does not affect the frequency ratios predicted for aberration patterns by a random breakage-and-rejoining model. The model is generally consistent with current data on many different types of aberrations, whether or not dsb usually occur in cryptic clusters. For a Revell-type exchange model, however, the predictions do depend on clustering configurations; they gradually approach the predictions of the breakage-and-rejoining model as average cluster multiplicity increases. The model is consistent with the data, for example with the ratio of visibly complex to apparently simple aberrations, only if there is considerable dsb clustering even at low-LET, with approximately 1.5 or more reactive dsb per cluster on average.

The immunolocalization and divergent roles of phosducin and phosducin-like protein in the retina.

PURPOSE: These investigations were undertaken to compare and contrast the roles of phosducin and phosducin-like protein in the retina. METHODS: Phosducin and phosducin-like protein were compared in an in vitro assay measuring their inhibition of transducin binding to light-activated rhodopsin. The two proteins were localized within the retina by immunoblot analyses and immunocytochemistry using affinity-purified antibodies with high specificity for each of the two homologs. The sensitivity of phosducin-like protein to phosphorylation was probed using in vitro protein kinase reactions. RESULTS: Phosducin and phosducin-like protein were found to have similar, though not identical, transducin inhibiting activity in vitro. These two proteins were found to be localized dissimilarly within the retina, with spatial overlap limited to the inner segments of the photoreceptors. Phosducin is found exclusively in photoreceptor cells, including the synaptic and nuclear layers, while phosducin-like protein is found throughout the inner retinal layers, most abundantly in the bipolar cells of the inner nuclear layer. Phosducin-like protein is not efficiently phosphorylated by the protein kinases tested, indicating that its regulation differs from that of phosducin. CONCLUSIONS: It appears that phosducin and phosducin-like protein play distinct roles in the retina. While phosducin is likely to be important in feedback regulation of the visual signal, such as in light adaptation, phosducin-like protein probably has little if any function in the phototransduction cascade. Phosducin-like protein may have a role in regulating the processing of visual signals by the neural cells of the inner retina.

An investigation of LET 'finger-prints' in Tradescantia.

Chromosome-type aberration scores, obtained in the Tradescantia microspore system following exposure to radiation, have been extracted from all significant papers since 1940, and augmented with unpublished results from this laboratory. The data include aberrations produced by X-, gamma-rays and neutrons, and cover an enormous range of qualities, doses, dose-rates and ambient gas conditions. Two proposed LET 'finger-print' ratios were examined: Dicentrics/Centric-rings (D/R or F-ratio) and Total deletions/Dicentrics+Centric-rings. There was no significant effect of LET on either of these. Further, D/R was independent of dose, whereas Deletions/D+R showed, as expected, a positive dose effect. In a very small subset of experiments, sizes of interstitial deletions had been recorded, and a significant reduction in modal size was observed for neutrons compared to X-rays. Even if this observation is confirmed by future work, it will not provide a usable 'finger-print' for long-term studies since, being acentric, deletions are rapidly eliminated from a dividing cell population.