The unusual γ-ray burst GRB 101225A from a helium star/neutron star merger at redshift 0.33.

Long γ-ray bursts (GRBs) are the most dramatic examples of massive stellar deaths, often associated with supernovae. They release ultra-relativistic jets, which produce non-thermal emission through synchrotron radiation as they interact with the surrounding medium. Here we report observations of the unusual GRB 101225A. Its γ-ray emission was exceptionally long-lived and was followed by a bright X-ray transient with a hot thermal component and an unusual optical counterpart. During the first 10 days, the optical emission evolved as an expanding, cooling black body, after which an additional component, consistent with a faint supernova, emerged. We estimate its redshift to be z = 0.33 by fitting the spectral-energy distribution and light curve of the optical emission with a GRB-supernova template. Deep optical observations may have revealed a faint, unresolved host galaxy. Our proposed progenitor is a merger of a helium star with a neutron star that underwent a common envelope phase, expelling its hydrogen envelope. The resulting explosion created a GRB-like jet which became thermalized by interacting with the dense, previously ejected material, thus creating the observed black body, until finally the emission from the supernova dominated. An alternative explanation is a minor body falling onto a neutron star in the Galaxy.

Relativistic jet activity from the tidal disruption of a star by a massive black hole.

Supermassive black holes have powerful gravitational fields with strong gradients that can destroy stars that get too close, producing a bright flare in ultraviolet and X-ray spectral regions from stellar debris that forms an accretion disk around the black hole. The aftermath of this process may have been seen several times over the past two decades in the form of sparsely sampled, slowly fading emission from distant galaxies, but the onset of the stellar disruption event has not hitherto been observed. Here we report observations of a bright X-ray flare from the extragalactic transient Swift J164449.3+573451. This source increased in brightness in the X-ray band by a factor of at least 10,000 since 1990 and by a factor of at least 100 since early 2010. We conclude that we have captured the onset of relativistic jet activity from a supermassive black hole. A companion paper comes to similar conclusions on the basis of radio observations. This event is probably due to the tidal disruption of a star falling into a supermassive black hole, but the detailed behaviour differs from current theoretical models of such events.

Deep neuromuscular block improves the surgical conditions for laryngeal microsurgery.

BACKGROUND: Adequate neuromuscular block is required throughout laryngeal microsurgery. We hypothesized that the surgical conditions would improve under a deeper level of rocuronium-induced neuromuscular block. METHODS: Seventy-two patients undergoing laryngeal microsurgery were randomly allocated to either the 'post-tetanic counts 1-2' (PTC1-2) group or the 'train-of-four counts 1-2' (TOFcount1-2) group according to the level of neuromuscular block used. Two different doses of rocuronium (1.2 or 0.5 mg kg(-1)) were used after anaesthetic induction, and two respective targets of neuromuscular block (post-tetanic counts ≤2 or train-of-four count of 1 or 2) were used. Surgical conditions were assessed by the surgeon using a five-point rating scale (extremely poor/poor/acceptable/good/optimal), and clinically acceptable surgical conditions were defined as those which were rated acceptable, good, or optimal. The occurrence of vocal cord movement and postoperative adverse events was assessed. RESULTS: The surgical conditions were significantly different between the PTC1-2 and TOFcount1-2 groups (extremely poor/poor/acceptable/good/optimal: 0/2/1/7/26 and 3/10/2/14/7, respectively, P<0.001). The incidence of clinically acceptable surgical conditions was significantly higher in the PTC1-2 group than in the TOFcount1-2 group (94 vs 64%, P=0.003). The percentage of patients who exhibited vocal cord movement was significantly lower in the PTC1-2 group than in the TOFcount1-2 group (3 vs 39%, P<0.001). The incidence of postoperative adverse events was not significantly different except for the less frequent occurrence of mouth dryness in the PTC1-2 group (P=0.035). CONCLUSIONS: Deep neuromuscular block (post-tetanic count of 1-2) surgical conditions in patients undergoing laryngeal microsurgery improves. CLINICAL TRIAL REGISTRATION: NCT01980069.

An assessment of oropharyngeal airway position using a fibreoptic bronchoscope.

Selecting the appropriate oropharyngeal airway for safe and effective airway management is important in clinical practice. In this prospective observational study, we examined the position of the distal end of oropharyngeal airways using a fibreoptic bronchoscope. We enrolled 149 adults (72 men and 77 women). The correct airway size was determined by inserting four adult sizes Guedel airway (Hudson RCI; Teleflex Medical, Research Triangle, Park, NC, USA) (sizes 8, 9, 10 and 11) sequentially in anaesthetised patients. The 'best fit' airway was size 10 in 45 (62%) men, and size 9 in 58 (75%) women. However, when these airways were inserted, the distal end of the airway either touched or passed beyond the epiglottis tip in 20 (27%) men and six (8%) women, respectively. When a size-9 airway was inserted in men and a size-8 airway inserted in women, the distal ends were obstructed by the tongue in three (2%) patients. In conclusion, a size-9 airway in men and a size-8 airway in women are the most acceptable sizes for adults of average height.

Observation of the hybridization gap and Fano resonance in the Kondo lattice URu2Si2.

The nature of the second-order phase transition that occurs in URu2Si2 at 17.5 K remains puzzling despite intensive research. A key question emerging in the field is whether a hybridization gap between the renormalized bands can be identified as the "hidden" order parameter. We report on the measurement of a hybridization gap in URu2Si2 employing a spectroscopic technique based on quasiparticle scattering. The differential conductance exhibits an asymmetric double-peak structure, a clear signature for a Fano resonance in a Kondo lattice. The hybridization gap opens well above 17.5 K, indicating that it is not the hidden order parameter. Our results put stringent constraints on the origin of the hidden order transition in URu2Si2 and demonstrate that quasiparticle scattering spectroscopy can probe the band renormalizations in a Kondo lattice via detection of a novel type of Fano resonance.

Smooth emergence in men undergoing nasal surgery: the effect site concentration of remifentanil for preventing cough after sevoflurane-balanced anaesthesia.

BACKGROUND: Suppression of cough during emergence after nasal surgery is practical to avoid bleeding from the surgical site. Previously recommended effect-site concentration of remifentanil for preventing cough during emergence is restricted to female patients undergoing thyroid surgery, so we evaluated effective effect-site concentration of remifentanil for preventing cough during emergence for men undergoing nasal surgery. METHODS: Twenty-four American Society of Anesthesiologists (ASA) physical status I or II, between the ages of 18 and 60 years old, non-smoker male patients undergoing nasal surgery were enrolled in this study. The effective effect-site concentration for 50% of patients (EC(50) ) and and that for 95% of patients (EC(95) ) of remifentanil for preventing cough were determined by Dixon's up-and-down method and by isotonic regression method with a bootstrapping approach. Haemodynamic variables were compared in patients with cough and without cough during emergence. RESULTS: The EC(50) of remifentanil for preventing cough during emergence by Dixon's method was 2.17 ng/ml [standard deviation (SD) 0.38]. The estimated EC(50) and EC(95) of remifentanil using isotonic regression model with a bootstrapping approach were 2.35 ng/ml [95% confidence interval (CI) 1.89-2.66] and 2.94 ng/ml (95% CI 2.83-2.97), respectively. Mean arterial pressure and heart rate were significantly higher in patients with cough during emergence. Three out of 13 patients that received more than 2.5 ng/ml of remifentanil experienced a brief episode of bradypnea. CONCLUSIONS: The EC(95) of remifentanil for preventing cough in men after nasal surgery is 2.94 ng/ml. Remifentanil target-controlled infusion with established effect-site concentration could suppress cough in men during emergence from sevoflurane anaesthesia after nasal surgery, though reversible respiratory depression might arise at high effect-site concentration of remifentanil.

Comparison between the SCL-90-R and MMPI in TMD patients with psychological problems.

OBJECTIVE: The aim of the study was to investigate the relationships between the Symptom Checklist-90-Revision (SCL-90-R) and the Minnesota Multiphasic Personality Inventory (MMPI) in temporomandibular disorders (TMD) patients with psychological problems. MATERIALS AND METHODS: Subjective symptoms, objective signs, and psychological characteristics of 36 TMD patients with psychological problems were analyzed. The symptom severity index (SSI) and craniomandibular index (CMI) were used to assess subjective symptoms and objective signs of patients with TMD, respectively. The SCL-90-R and MMPI were used for psychological evaluation. RESULTS: The SSI was not significantly correlated with the CMI in TMD patients with psychological problems, and these indices displayed significant correlations with the SCL-90-R and MMPI in several selected subscales. The results of SCL-90-R had a limited relationship with those of MMPI in these patients. Based on the MMPI diagnosis, the SCL-90-R somatization subscale showed moderate to high sensitivity and specificity, but the SCL-90-R depression subscale showed moderate to low sensitivity and specificity. CONCLUSIONS: Considering the limited relationship between the SCL-90-R and MMPI in TMD patients with psychological problems, more comprehensive psychological tests are recommended when clinicians suspect patients with TMD of having accompanying psychological problems.

Phosphodiesterase inhibition by naloxone augments the inotropic actions of beta-adrenergic stimulation.

BACKGROUND: In a shock state, naloxone generates the cardiovascular pressor effect by displacing the endogenous opiate-like peptide beta-endorphin, resulting in restoration of the normal response to catecholamines. In addition to this opioid antagonistic effect, the non-opiate receptor-mediated effect has also been proposed. The aim of this study was to define the mechanism of non-opiate receptor-mediated action of naloxone. METHODS: In guinea-pig ventricular tissues, cumulative concentration-response curves for isoproterenol as well as for forskolin and 3-isobutylmethylxanthine (IBMX) were obtained by increasing the concentration stepwise. To assess the effect on the phosphodiesterase (PDE), the effects of naloxone on contractile forces induced by isoproterenol (0.05 microM) in the presence of IBMX, cilostamide (a PDE III inhibitor), or rolipram (a PDE IV inhibitor) were observed. Naloxone-induced changes in cAMP production by isoproterenol both in the absence and in the presence of IBMX were measured. Naloxone-induced changes in cAMP production by forskolin in the presence of IBMX were also measured. RESULTS: Naloxone (30 microM) produced a leftward shift of the isoproterenol concentration-response curve (0.01-2 microM) without changing the maximal response. Forskolin (0.5-10 microM) produced a concentration-dependent increase in contractile forces. Naloxone increased the maximal inotropic response of forskolin. Naloxone showed no effect on the IBMX concentration-response curve. In the presence of IBMX (200 microM), naloxone did not alter the contractions evoked by isoproterenol or forskolin. Whereas naloxone increased contractile forces significantly (approximately 25%) more than that of isoproterenol in the presence of rolipram, no alteration of contractile forces in the cilostamide-incubated muscles was observed. Naloxone caused a concentration-related increase of cAMP in the absence of IBMX, but caused no change in its presence. CONCLUSIONS: The enhancement of myocardial contractility by naloxone in the presence of stimulation of adenylyl cyclase activity appears to be mediated by inhibition of PDE, specifically PDE III.

Andreev reflection and order parameter symmetry in heavy-fermion superconductors: the case of CeCoIn(5).

We review the current status of Andreev reflection spectroscopy on the heavy fermions, mostly focusing on the case of CeCoIn(5), a heavy-fermion superconductor with a critical temperature of 2.3 K. This is a well-established technique to investigate superconducting order parameters via measurements of the differential conductance from nanoscale metallic junctions. Andreev reflection is clearly observed in CeCoIn(5) as in other heavy-fermion superconductors. Considering the large mismatch in Fermi velocities, this observation seemingly appears to disagree with the Blonder-Tinkham-Klapwijk (BTK) theory. The measured Andreev signal is highly reduced to the order of maximum ∼13% compared to the theoretically predicted value (100%). The background conductance exhibits a systematic evolution in its asymmetry over a wide temperature range from above the heavy-fermion coherence temperature down to well below the superconducting transition temperature. Analysis of the conductance spectra using the extended BTK model provides a qualitative measure for the superconducting order parameter symmetry, which is determined to be the d(x(2)-y(2)) wave in CeCoIn(5). It is found that existing models do not quantitatively account for the data, which we attribute to the intrinsic properties of the heavy fermions. A substantial body of experimental data and extensive theoretical analysis point to the existence of two-fluid components in CeCoIn(5) and other heavy-fermion compounds. A phenomenological model is proposed employing a Fano interference effect between two conductance channels in order to explain both the conductance asymmetry and the reduced Andreev signal. This model appears plausible not only because it provides good fits to the data but also because it is highly likely that the electrical conduction occurs via two channels, one into the heavy-electron liquid and the other into the conduction electron continuum. Further experimental and theoretical investigations will shed new light on the mechanism of how the coherent heavy-electron liquid emerges out of the Kondo lattice, a prototypical strongly correlated electron system. Unresolved issues and future directions are also discussed.

Intra-operative warming with a forced-air warmer in preventing hypothermia after tourniquet deflation in elderly patients.

This randomized, single-blind study aimed to explore the effects of intra-operative warming with a forced-air warmer in the prevention of hypothermia after tourniquet deflation in elderly patients undergoing unilateral total knee replacement arthroplasty under general anaesthesia. Patients were randomized to receive either intra-operative warming using a forced-air warmer with an upper body blanket (warming group; n = 12) or no intra-operative warming (nonwarming group; n = 12). Oesophageal temperature was measured as core body temperature. At 30 min following tourniquet inflation, the core body temperature started to increase in the warming group whereas it continued to drop in the non-warming group. This difference was statistically significant. The final core body temperature after tourniquet deflation was significantly higher in the warming group (mean +/- SD 36.1 +/- 0.2 degrees C) than in the non-warming group (35.4 +/- 0.3 degrees C). Intra-operative forced-air warming increased the core body temperature before tourniquet deflation and prevented subsequent hypothermia in elderly patients under general anaesthesia.

Design and construction of a point-contact spectroscopy rig with lateral scanning capability.

The design and realization of a cryogenic rig for point-contact spectroscopy measurements in the needle-anvil configuration is presented. Thanks to the use of two piezoelectric nano-positioners, the tip can move along the vertical (z) and horizontal (x) direction and thus the rig is suitable to probe different regions of a sample in situ. Moreover, it can also form double point-contacts on different facets of a single crystal for achieving, e.g., an interferometer configuration for phase-sensitive measurements. For the later purpose, the sample holder can also host a Helmholtz coil for applying a small transverse magnetic field to the junction. A semi-rigid coaxial cable can be easily added for studying the behavior of Josephson junctions under microwave irradiation. The rig can be detached from the probe and thus used with different cryostats. The performance of this new probe has been tested in a Quantum Design PPMS system by conducting point-contact Andreev reflection measurements on Nb thin films over large areas as a function of temperature and magnetic field.

Cocaine administered into the medial prefrontal cortex reinstates cocaine-seeking behavior by increasing AMPA receptor-mediated glutamate transmission in the nucleus accumbens.

One of the major determinants of reinstatement to cocaine use among human addicts is acute reexposure to the drug, which often precipitates cocaine craving and relapse. We used an animal model of cocaine relapse to determine the role of the glutamatergic pathway from the medial prefrontal cortex (mPFC) to the nucleus accumbens in the reinstatement of cocaine-seeking behavior after a cocaine priming injection. Rats were trained to self-administer cocaine intravenously on a second order schedule. Responding was extinguished subsequently by substituting saline for cocaine. During subsequent reinstatement sessions, drug-seeking behavior was assessed after noncontingent priming injections. Results indicated that reinstatement induced by a systemic cocaine injection was blocked by intra-mPFC administration of the dopamine antagonist flupenthixol. Consistent with this finding, administration of cocaine directly into the mPFC reinstated cocaine-seeking behavior. Administration of cocaine into the nucleus accumbens also reinstated drug seeking, whereas microinjection of cocaine into the neostriatum or lateral septum did not. Reinstatement of cocaine seeking induced by intra-mPFC cocaine was blocked by administration of the AMPA receptor antagonist CNQX into the nucleus accumbens. Administration of the NMDA receptor antagonist AP-5 into the nucleus accumbens had variable effects on reinstatement induced by intra-mPFC cocaine in that AP-5 had no effect in some animals but augmented reinstatement in others. Subsequent experiments showed that intra-accumbal microinjection of AP-5 alone dose-dependently reinstated cocaine seeking. These data indicate that the glutamatergic pathway from the mPFC to the nucleus accumbens plays an important role in cocaine priming-induced reinstatement of drug seeking. Moreover, the present results demonstrate that AMPA and NMDA receptors in the nucleus accumbens have opposing roles in the reinstatement of cocaine-seeking behavior.

A reexamination of the role of GABA in the mammalian suprachiasmatic nucleus.

Three independent electrophysiological approaches in hypothalamic slices were used to test the hypothesis that gamma-amino butyric acid (GABA)A receptor activation excites suprachiasmatic nucleus (SCN) neurons during the subjective day, consistent with a recent report. First, multiple-unit recordings during either the subjective day or night showed that GABA or muscimol inhibited firing activity of the SCN population in a dose-dependent manner. Second, cell-attached recordings during the subjective day demonstrated an inhibitory effect of bath- or microapplied GABA on action currents of single SCN neurons. Third, gramicidin perforated-patch recordings showed that bicuculline increased the spontaneous firing rate during the subjective day. Therefore, electrophysiological data obtained by three different experimental methods provide evidence that GABA is inhibitory rather than excitatory during the subjective day.

Mechanical and electrophysiological effects of protamine on isolated ventricular myocardium: evidence for calcium overload.

OBJECTIVE: The aim was to examine in vitro the cellular cardiac effects of protamine, the cationic polypeptide employed to reverse heparin anticoagulation, and to define its mechanisms of action. METHODS: Isometric contractile force and action potential characteristics after rest and at frequencies up to 3 Hz were recorded in guinea pig ventricular papillary muscle. The actions of protamine (10-300 micrograms.ml-1) were compared to those of heparin (10, 30 units.ml-1), and to heparin (10 units.ml-1) neutralised with equivalent (100 micrograms.ml-1) or excess (200 micrograms.ml-1) protamine. The effects of protamine were also examined using (1) muscle rapid cooling contractures (RCC) to assess intracellular Ca2+ stores and (2) the whole cell voltage clamp method to evaluate K+ and Ca2+ currents of isolated ventricular myocytes. RESULTS: Protamine (100-300 micrograms.ml-1) depressed contractions by 35-65% at 3 Hz, whereas contractions were enhanced by 150-500% at lower rates (resting state, 0.5 Hz), with a concomitant rise in resting force. Protamine caused a resting depolarisation from -90 to -76 mV and depressed action potential amplitude. In contrast, heparin altered contractile or action potential characteristics minimally. In 26 mM K+ solution with 0.1 microM isoprenaline, 30-300 micrograms.ml-1 protamine caused dose dependent depression of late peaking force development and slow action potential prolongation. After 15 min rest, when RCC were not normally elicited, rest RCC became prominent in 100-300 micrograms.ml-1 protamine. Effects of heparin with 100 micrograms.ml-1 excess protamine were similar to those of 100 micrograms.ml-1 protamine alone. Voltage clamp of isolated myocytes revealed that 10 micrograms.ml-1 protamine irreversibly decreased current through inwardly rectifying K+ channels (IK1), increased leakage current, and decreased inward Ca2+ current (ICa). CONCLUSIONS: The loss of the normal force-frequency relation, partial depolarisation, rise in resting tension, and appearance of rested state rapid cooling contractures suggest that unbound protamine can lead to excess intracellular Ca2+, mediated by an alteration in membrane ionic conductances.

Two-step fabrication technique of gold tips for use in point-contact spectroscopy.

For a successful point-contact spectroscopy (PCS) measurement, metallic tips of proper shape and smoothness are essential to ensure the ballistic nature of a point-contact junction. Until recently, the fabrication of Au tips suitable for use in point-contact spectroscopy has remained more of an art involving a trial and error method rather than an automated scientific process. To address these issues, we have developed a technique with which one can prepare high quality Au tips reproducibly and systematically. It involves an electronic control of the driving voltages used for an electrochemical etching of a gold wire in a HCl-glycerol mixture or a HCl solution. We find that a stopping current, below which the circuit is set to shut off, is a single very important parameter to produce an Au tip of desired shape. We present detailed descriptions for a two-step etching process for Au tips and also test results from PCS measurements using them.

Specific antagonists for the myotropic action of angiotensin II and angiotensin I on the isolated rat colon.

Substitutions of 8 phenylalanine with L-alanine and D-phenylalanine abolish the myotropic action of the angiotensin II (AT(II)) analogues and confer inhibitory properties on the molecule. [8-L-Ala]-AT(II) and [8-D-Phe]-AT(II) antagonize specifically the myotropic action of AT(II) and angiotensin I (AT(I)) on the rat colon, while the action of other myotropic agents (acetylcholine, 5 hydroxytryptamine) is not modified.

Characterization of angiotensin receptors in vascular and intestinal smooth muscles.

1. A series of analogues of angiotensin II (AT(II)) has been used in the present experiments to characterize receptors for AT(II) in intestinal (rat stomach strip, rat colon) and vascular (rabbit aorta) smooth muscles. Two types of compounds have been chosen: (a) agonists with reduced potency, in which 4-Tyr, 6-His or 7-Pro had been substituted with L-Ala or with Gly and 1-amino-cyclopentane carboxylic acid (Acpc) and (b) competitive antagonists (8-Gly-AT(II), 8-Leu-AT(II)).2. Replacement of 4-Tyr, 6-His and 7-Pro with L-Ala decreases the potency, but does not influence the maximum effect of the analogue, while substitution of the same residue with Gly and Acpc reduces both potency and maximum effect.3. Compounds showing full size maximum responses were chosen to establish the following order of potency on the three preparations: AT(II)>4-Phe-AT(II)>7-Ala-AT(II)>6-Ala-AT(II)>4-Ala-AT(II).4. The four derivatives of AT(II) were completely inactive on tissues desensitized with AT(II). The responses to 5-hydroxytryptamine, acetylcholine and noradrenaline were not significantly modified, except for the rat colon.5. pA(2) values for the two competitive antagonists against AT(II) and 4-Phe-AT(II) were estimated in the three preparations by the use of the cascade superfusion technique. For comparison, pA(2) values were also estimated in rat stomach strips and rabbit aortas suspended in a normal organ bath, according to the method of Schild (1947). The similarities of the pA(2) values obtained in two series of experiments indicate that (a) the cascade superfusion technique is suitable for this type of study and (b) the receptor for AT(II) in the three tissues may be the same.6. It is suggested that receptors for AT(II) in intestinal and vascular smooth muscles may be the same, because (a) the order of potency of various agonists follows the same pattern, (b) the agonists are inactive on tissues desensitized with AT(II), (c) pA(2) values for competitive antagonists are similar in the three preparations.

Activation of the neutrophil respiratory burst requires both intracellular and extracellular calcium.

Activation of neutrophil oxidases, including NADPH oxidase, is Ca2+ dependent. The aim of this study was to determine the roles of intra- and extracellular Ca2+, leading to generation of the respiratory burst, as monitored by luminol-dependent chemiluminescence (CL). All results were recorded as integrals (millivolt.min) and compared by a two-tail Student's t test. Preincubation of cells with chelators of intra- or extracellular Ca2+ inhibited N-Formyl-Met-Leu-Phe (FMLP)-stimulated burst activity (p < 0.01). In contrast, stimulation by phorbol myristate acetate (PMA), while inhibited by extracellular Ca2+ chelation with EGTA (p < 0.001), was potentiated by intracellular Ca2+ chelation with BAPTA (p < 0.01). This suggests that the protein kinase C (PKC)-mediated burst may be diminished by intracellular Ca(2+)-dependent phosphatase. A selective inhibitor of tyrosine phosphatase, sodium vanadate, potentiated CL generation by both FMLP and PMA, indicating a dominant phosphatase activation with transiently increased Ca2+, masking the kinase-mediated respiratory burst. The selective inhibitors of PKC or tyrosine kinase prevented PMA and vanadate/PMA stimulation (p < 0.005). Furthermore, the putative Ca2+ channel agonists glutamate (10(-5)M) and N-methyl-D-aspartate (NMDA) (10(-5)M) alone failed to influence CL output, but produced marked potentiation following pre-treatment with vanadate. Again this indicates a dominant activation of phosphatase triggered by the glutamate-mediated Ca2+ influx, so masking the kinase-dependent NADPH oxidase activity. A competitive antagonist of NMDA, AP7, significantly decreased vanadate-mediated CL in an EGTA-sensitive manner (p < 0.001). The data confirm a requirement for intra- and extracellular Ca2+ in neutrophil respiratory burst activation via the kinase/phosphatase cycle, and an agonist effect by NMDA within the Ca2+ cascade mechanism.

Influence of heat inactivation of human serum on the opsonization of Streptococcus mutans.

Phagocytosis of bacteria, such as Streptococcus mutans, is important to host defense. One mechanism by which phagocytosis can be enhanced is by antibody or complement-mediated opsonization of bacteria. Many studies utilize opsonization of bacteria to enhance a cellular response, but little information has been found examining methodology or validity of the opsonization process following the denaturization of the serum. Human serum was inactivated by heat in order to disrupt the classical and alternative pathways of the complement cascade. S. mutans isolated from human subjects were opsonized with heat-inactivated human serum before exposing them to viable neutrophils in vitro. Luminol-dependent chemiluminescence (CL) was used to measure neutrophil activation. Human serum used to opsonize the bacteria was denatured by incubation at 57 degrees C for intervals of 30 and 60 min to inactivate complement. The results from the opsonization data indicated that there was significantly increased CL with 60-min inactivation of the serum (34% increase in mean integration mV.min; p < or = 0.05) over the nonopsonized control. This indicated a successful opsonization of the bacteria. In addition, the data demonstrate that the inactivation of serum requires a minimum of 60 min at 57 degrees C to disrupt the complement cascade, while 30- and 15-min inactivations produced no significant increase in CL activity over the control. Standard sandwich ELISA assays, detecting complement binding to S. mutans, confirmed successful heat inactivation of serum showing a significant decrease (p < or = 0.001) in complement binding to S. mutans after 30 min, but could not explain the increased CL response after 60-min heat deactivation of the serum.

Chemical libraries towards protein kinase inhibitors.

Over 500 human protein kinases identified to date are susceptible to play crucial roles in the regulation of many signal transduction pathways, making them significant drug discovery targets. However, their active sites share a high level of similarity, which constitutes a major challenge in the finding of selective and safe inhibitors. In order to meet this challenge, whether via traditional or alternative approaches, the use of chemical libraries to find either unknown natural ligands or specific inhibitors of particular kinases is more important than ever. This review briefly summarizes the recent literature on such libraries of peptides, natural product analogues, and small molecules. Significant chemical scaffolds, some synthetic routes particularly on solid-phase support, and computational tools employed for the efficient design of both selective and bioavailable inhibitors are highlighted.