Prohormone convertase 2 (PC2) null mice have increased mu opioid receptor levels accompanied by altered morphine-induced antinociception, tolerance and dependence.

Chronic morphine treatment increases the levels of prohormone convertase 2 (PC2) in brain regions involved in nociception, tolerance and dependence. Thus, we tested if PC2 null mice exhibit altered morphine-induced antinociception, tolerance and dependence. PC2 null mice and their wild-type controls were tested for baseline hot plate latency, injected with morphine (1.25-10mg/kg) and tested for antinociception 30min later. For tolerance studies, mice were tested in the hot plate test before and 30min following morphine (5mg/kg) on day 1. Mice then received an additional dose so that the final dose of morphine was 10mg/kg on this day. On days 2-4, mice received additional doses of morphine (20, 40 and 80mg/kg on days 1, 2, 3, and 4, respectively). On day 5, mice were tested in the hot plate test before and 30min following morphine (5mg/kg). For withdrawal studies, mice were treated with the escalating doses of morphine (10, 20, 40 and 80mg/kg) for 4days, implanted with a morphine pellet on day 5 and 3 days later injected with naloxone (1mg/kg) and signs of withdrawal were recorded. Morphine dose-dependently induced antinociception and the magnitude of this response was greater in PC2 null mice. Tolerance to morphine was observed in wild-type mice and this phenomenon was blunted in PC2 null mice. Withdrawal signs were also reduced in PC2 null mice. Immunohistochemical studies showed up-regulation of the mu opioid receptor (MOP) protein expression in the periaqueductal gray area, ventral tegmental area, lateral hypothalamus, medial hypothalamus, nucleus accumbens, and somatosensory cortex in PC2 null mice. Likewise, naloxone specific binding was increased in the brains of these mice compared to their wild-type controls. The results suggest that the PC2-derived peptides may play a functional role in morphine-induced antinociception, tolerance and dependence. Alternatively, lack of opioid peptides led to up-regulation of the MOP and altered morphine-induced antinociception, tolerance and dependence.

Clinical characteristics, hospital outcome and prognostic factors of patients with ventilator-related pneumothorax.

BACKGROUND: Mechanical ventilation is a common cause of iatrogenic pneumothorax in intensive care units (ICU). Most of the patients with ventilator-related pneumothorax (VRP) have underlying lung diseases and is associated with increased morbidity and mortality. The prognostic factors of VRP are not clear. The objective of this study was to find the possible prognostic factors. METHODS: Analysis of retrospectively collected data of patients with pneumothorax induced by mechanical ventilation. Data were obtained concerning demographics, acute physiology and chronic health evaluation (APACHE) II score, organ failure, underlying diseases, interval between the start of mechanical ventilation and pneumothorax, arterial blood gas, respiratory parameters and patient outcomes. RESULTS: One hundred and twenty-four patients with VRP were included for analysis. The incidence rate of VRP was 0.4% (124/31,660), and the mortality rate was 77.4%. The patients with VRP had higher hospital mortality rate than that of mechanically ventilated patients without pneumothorax (77.4% vs. 13.7%, P<0.001) or patient with procedure-related pneumothorax (77.4% vs. 29.4%, P<0.001). Most cases of VRP occurred in the early phase of mechanical ventilation, and 8.9% of the patients had a later episode of pneumothorax on the opposite lung. The interval between two episodes of VRP was short, at a median time of 2 days. Cox regression analysis showed that tension pneumothorax (P=0.001), PaO2/FiO2<200 (P=0.002), and APACHE II score (P=0.008) were significantly associated with death. CONCLUSION: VRP patients with tension pneumothorax or PaO2/FiO2<200 had a higher risk of death. APACHE II scores were associated with mortality in the VRP patients with PaO2/FiO2≥200 mmHg.

Spontaneous tonsillar haemorrhage due to von Willebrand's disease.

OBJECTIVE: We report a case of spontaneous tonsillar haemorrhage due to underlying von Willebrand's disease, without symptoms or signs of infection. METHOD: Case report and literature review of reported causes of spontaneous tonsillar haemorrhage. CASE REPORT: Spontaneous tonsillar haemorrhage is uncommon. Acute bacterial tonsillitis, peri-tonsillar abscess, infectious mononucleosis and idiopathic causes have been reported. Two cases of tonsillitis with underlying bleeding disorders have been reported. We present the first reported case of spontaneous tonsillar haemorrhage secondary to von Willebrand's disease, without evidence of tonsillitis. CONCLUSION: Spontaneous tonsillar haemorrhage is usually related to infection of the tonsils or peri-tonsillar space. An underlying bleeding disorder should be suspected if no evidence of infection is found.

Optimal combination of nested clusters by a greedy approximation algorithm.

Given a set of clusters, we consider an optimization problem which seeks a subset of clusters that maximizes the microaverage F-measure. This optimal value can be used as an evaluation measure of the goodness of clustering. For arbitrarily overlapping clusters, finding the optimal value is NP-hard. We claim that a greedy approximation algorithm yields the global optimal solution for clusters that overlap only by nesting. We present a mathematical proof of this claim by induction. For a family of n clusters containing a total of N objects, this algorithm has an {\rm O}(n;{2}) time complexity and O(N) space complexity.

Effect of horizontal position of the computer keyboard on upper extremity posture and muscular load during computer work.

The distance of the keyboard from the edge of a work surface has been associated with hand and arm pain; however, the variation in postural and muscular effects with the horizontal position have not been explicitly explored in previous studies. It was hypothesized that the wrist approaches more of a neutral posture as the keyboard distance from the edge of table increases. In a laboratory setting, 20 adults completed computer tasks using four workstation configurations: with the keyboard at the edge of the work surface (NEAR), 8 cm from the edge and 15 cm from the edge, the latter condition also with a pad that raised the work surface proximal to the keyboard (FWP). Electrogoniometers and an electromagnetic motion analysis system measured wrist and upper arm postures and surface electromyography measured muscle activity of two forearm and two shoulder muscles. Wrist ulnar deviation decreased by 50% (4 degrees ) as the keyboard position moved away from the user. Without a pad, wrist extension increased by 20% (4 degrees ) as the keyboard moved away but when the pad was added, wrist extension did not differ from that in the NEAR configuration. Median values of wrist extensor muscle activity decreased by 4% maximum voluntary contraction for the farthest position with a pad (FWP). The upper arm followed suit: flexion increased while abduction and internal rotation decreased as the keyboard was positioned further away from the edge of the table. In order to achieve neutral postures of the upper extremity, the keyboard position in the horizontal plane has an important role and needs to be considered within the context of workstation designs and interventions.

Effects of single D-amino acid substitutions on disruption of beta-sheet structure and hydrophobicity in cyclic 14-residue antimicrobial peptide analogs related to gramicidin S.

Gramicidin S (GS) is a 10-residue cyclic beta-sheet peptide with lytic activity against the membranes of both microbial and human cells, i.e. it possesses little to no biologic specificity for either cell type. Structure-activity studies of de novo-designed 14-residue cyclic peptides based on GS have previously shown that higher specificity against microbial membranes, i.e. a high therapeutic index (TI), can be achieved by the replacement of a single L-amino acid with its corresponding D-enantiomer [Kondejewski, L.H. et al. (1999) J. Biol. Chem. 274, 13181]. The diastereomer with a D-Lys substituted at position 4 caused the greatest improvement in specificity vs. other L to D substitutions within the cyclic 14-residue peptide GS14, through a combination of decreased peptide amphipathicity and disrupted beta-sheet structure in aqueous conditions [McInnes, C. et al. (2000) J. Biol. Chem. 275, 14287]. Based on this information, we have created a series of peptide diastereomers substituted only at position 4 by a D- or L-amino acid (Leu, Phe, Tyr, Asn, Lys, and achiral Gly). The amino acids chosen in this study represent a range of hydrophobicities/hydrophilicities as a subset of the 20 naturally occurring amino acids. While the D- and L-substitutions of Leu, Phe, and Tyr all resulted in strong hemolytic activity, the substitutions of hydrophilic D-amino acids D-Lys and D-Asn in GS14 at position 4 resulted in weaker hemolytic activity than in the L-diastereomers, which demonstrated strong hemolysis. All of the L-substitutions also resulted in poor antimicrobial activity and an extremely low TI, while the antimicrobial activity of the D-substituted peptides tended to improve based on the hydrophilicity of the residue. D-Lys was the most polar and most efficacious substitution, resulting in the highest TI. Interestingly, the hydrophobic D-amino acid substitutions had superior antimicrobial activity vs. the L-enantiomers although substitution of a hydrophobic D-amino acid increases the nonpolar face hydrophobicity. These results further support the role of hydrophobicity of the nonpolar face as a major influence on microbial specificity, but also highlights the importance of a disrupted beta-sheet structure on antimicrobial activity.

Increased endothelin-induced Ca2+ signaling, tyrosine phosphorylation, and coronary artery disease in diabetic dyslipidemic Swine are prevented by atorvastatin.

Endothelin-1 (ET-1) signaling mechanisms have been implicated in the pathogenesis of excess coronary artery disease in diabetic dyslipidemia. We hypothesized that in diabetic dyslipidemia ET-1-induced coronary smooth muscle calcium (Ca2+m) and tyrosine phosphorylation would be increased, and the lipid lowering agent, atorvastatin, would inhibit these increases. Male Yucatan miniature swine groups were treated for 20 weeks: normal low-fat fed control, high-fat/cholesterol fed (hyperlipidemic), hyperlipidemic made diabetic with alloxan (diabetic dyslipidemic), and diabetic dyslipidemic treated with atorvastatin (atorvastatin-treated). Blood glucose values were 5-fold greater in diabetic dyslipidemic and atorvastatin-treated versus control and hyperlipidemic. Total and low-density lipoprotein (LDL) plasma cholesterol in hyperlipidemic, diabetic dyslipidemic, and atorvastatin-treated were approximately 5-fold greater than control. Intravascular ultrasound detectable coronary disease and hypertriglyceridemia were only observed in diabetic dyslipidemic and were abolished by atorvastatin. In freshly isolated cells, the Ca2+m response to ET-1 in diabetic dyslipidemic was greater than in control, hyperlipidemic, and atorvastatin-treated groups. Selective ET-1 receptor antagonists showed in the control group that the ETB subtype inhibits ETA regulation of Ca2+m. There was almost a complete switch of receptor subtype regulation of Ca2+m from largely ETA in control to an increased inhibitory interaction between ETA and ETB in hyperlipidemic and diabetic dyslipidemic groups, such that neither ETA nor ETB antagonist alone could block the ET-1-induced Ca2+m response. The inhibitory interaction was attenuated in the atorvastatin-treated group. In single cells, basal and ET-1-induced tyrosine phosphorylation in diabetic dyslipidemic were more than 3- and 6-fold greater, respectively, than in control, hyperlipidemic, and atorvastatin-treated. Attenuation by atorvastatin of coronary disease and ET-1-induced Ca2+m and tyrosine phosphorylation signaling with no change in cholesterol provides strong evidence for direct actions of atorvastatin and/or triglycerides on the vascular wall.

Investigation of a direct conversion flat panel imager for portal imaging.

Flat-panel based x-ray imaging is an emerging new technology that could be used to significantly improve the quality of on-line portal imagers. There are two types of flat panel imagers: direct and indirect conversion. Previous experimental work on flat panel detectors for portal imaging application used indirect-conversion imagers. In this paper, a direct-conversion amorphous-selenium flat panel imager is investigated for application in portal imaging. The imager has an active imaging area of 14 in. X 17 in., i.e., 3072 X 2560 pixels each with dimensions 139 microm X 139 microm. The spatial frequency dependent detective quantum efficiency of the imager has been measured for a 6 MV beam and found to be amongst the best area detectors investigated for on-line portal imaging. Effects of changing pixel size, as well as possible improvements to both the image quality and convenience of operation are discussed. Comparison with an indirect conversion flat panel imager is also included.

The effects of mastery training and explicit feedback on task design preference in a vocational setting.

The purpose of this study was to examine the effects of mastery training and explicit feedback on the selection behavior of two individuals with severe mental retardation across two different vocational task designs. The two design options were (a) a site-based (traditional) approach, and (b) a more efficient motion-economy based approach. The site-based design was developed from workshop-standard task analyses. The motion-economy design was developed by re-configuring the site-based design using principles of motion economy and resulted in less distance movement required for task completion. The study was comprised of four assessment phases: (a) no experience (i.e., participant had no experience with either site-based or motion-economy based designs), (b) after training (i.e., participant was trained to criterion level on both design options), (c) training and timer, and (d) training, timer, and work incentives. Selection behavior was affected by task efficiency, only when efficiency was made more salient by pairing task cues with work incentives.

Are trigger sequences essential in the folding of two-stranded alpha-helical coiled-coils?

The amino acid residues comprising the interface between strands of the coiled-coil motif are usually hydrophobic and make a major contribution to coiled-coil folding and stability. However, in some cases the presence of excellent hydrophobic residues at the coiled-coil interface is insufficient for folding. It has been proposed that a "consensus trigger sequence" exists that is necessary within the coiled-coil domains of various proteins to trigger folding. Therefore, in this study we designed a 31-residue hybrid sequence based on sequences from the two-stranded parallel coiled-coil domains of the yeast transcriptional activator GCN4 and the actin-bundling protein Dictyostelium discoideum cortexillin I. The hybrid and its analogs were studied by CD spectroscopy and analytical ultracentrifugation. The hybrid had stable residues in the core "a" and "d" positions in the 3-4 hydrophobic repeat, denoted (abcdefg)n, but did not have a consensus trigger sequence and did not possess appreciable secondary structure as determined by CD spectroscopy. The substitutions in the parent peptide were introduced at positions other than "a" and "d", altering a variety of interactions including alpha-helical propensity, interchain and intrachain electrostatics, and hydrophobicity. Although the substitutions did not bring the overall sequence in closer agreement to the consensus trigger sequence, they increased coiled-coil folding and stability. Therefore, our results suggest that the combination of stabilizing effects along a protein sequence is a more general indicator of protein folding in coiled-coils than the identification of a specific trigger sequence. We propose that surpassing a critical threshold stability value using any type or combination of stabilizing effects will allow coiled-coils to fold, in the absence of a specific trigger sequence per se.

Improving the capacity of complex-valued neural networks with a modified gradient descent learning rule.

Jankowski et al. proposed (1996) a complex-valued neural network (CVNN) which is capable of storing and recalling gray-scale images. The convergence property of the CVNN has also been proven by means of the energy function approach. However, the memory capacity of the CVNN is very low because they use a generalized Hebb rule to construct the connection matrix. In this letter, a modified gradient descent learning rule (MGDR) is proposed to enhance the capacity of the CVNN. The proposed technique is derived by applying gradient search over a complex error surface. Simulation shows that the capacity of CVNN with MGDR is greatly improved.

Relaxation of the stability condition of the complex-valued neural networks.

Jankuwski et al. (1996) have proposed a complex-valued neural network (CVNN) that is capable of storing and recalling gray-scale images. However, the weight matrix of the CVNN must be Hermitian with nonnegative diagonal entries in order to preserve the stability of the network. The Hermitian assumption poses difficulties in both physical realizations and practical applications of the networks. In this paper, a new stability condition is derived. The obtained result not only permits a little relaxation on the Hermitian assumption of the connection matrix, but also generalizes some existing results.

Chronic unloading by left ventricular assist device reverses contractile dysfunction and alters gene expression in end-stage heart failure.

BACKGROUND: Left ventricular (LV) assist devices (LVADs) can improve contractile strength and normalize characteristics of the Ca(2+) transient in myocytes isolated from failing human hearts. The purpose of the present study was to determine whether LVAD support also improves contractile strength at different frequencies of contraction (the force-frequency relationship [FFR]) of intact myocardium and alters the expression of genes encoding for proteins involved in Ca(2+) handling. METHODS AND RESULTS: The isometric FFRs of LV trabeculae isolated from 15 patients with end-stage heart failure were compared with those of 7 LVAD-supported patients and demonstrated improved contractile force at 1-Hz stimulation, with reversal of a negative FFR after LVAD implantation. In 20 failing hearts, Northern blot analysis for sarcoplasmic endoreticular Ca(2+)-ATPase subtype 2a (SERCA2a), the ryanodine receptor, and the sarcolemmal Na(+)-Ca(2+) exchanger was performed on LV tissue obtained before and after LVAD implantation. These paired data demonstrated an upregulation of all 3 genes after LVAD support. In tissue obtained from subsets of these patients, Western blot analysis was performed, and oxalate-supported Ca(2+) uptake by isolated sarcoplasmic reticular membranes was determined. Despite higher mRNA for all genes after LVAD support, only SERCA2a protein was increased. Functional significance of increased SERCA2a was confirmed by augmented Ca(2+) uptake by sarcoplasmic reticular membranes isolated from LVAD-supported hearts. CONCLUSIONS: LVAD support can improve contractile strength of intact myocardium and reverse the negative FFR associated with end-stage heart failure. The expression of genes encoding for proteins involved in Ca(2+) cycling is upregulated (reverse molecular remodeling), but only the protein content of SERCA2a is increased.

Vaccination of young lambs against infection with Nematodirus battus using gamma irradiated larvae.

Helminthologically naIve 6-week-old Suffolk lambs were given 1-3 doses of 20000 gamma-irradiated infective larvae (L3) of the nematode Nematodirus battus at weekly intervals. Following an anthelmintic drench they were challenged with 50000 viable L3 at 10 weeks of age. Nematode worm burdens 14 days post-challenge showed a significant (P<0.01) 66% reduction in the single vaccine dose group. The two and three dose groups had mean worm burdens which were 30 and 42% lower than controls, respectively, although these were not statistically significant. There was little measurable stimulation of the immune system in the vaccinated lambs, suggesting that the repeatedly dosed animals may have developed immunological unresponsiveness to the parasite.

An anteromedial internal jugular vein successfully cannulated using the assistance of ultrasonography.

The internal jugular vein usually is found either lateral or anterolateral to the carotid artery when it is cannulated for central vein access using external anatomical landmarks. We report a case in which the carotid artery was inadvertently punctured, but the right internal jugular vein could not be found. We used ultrasonic guidance to determine that the right internal jugular vein was anteromedial to the carotid artery. A figure showing the ultrasound of this rare anatomical variation is provided. The advantages and utility of ultrasound when used for the placement of internal jugular central vein catheters are reviewed.

Neonicotinoid insecticides: molecular features conferring selectivity for insect versus mammalian nicotinic receptors.

The favorable selective toxicity of neonicotinoid insecticides (represented here by imidacloprid, thiacloprid, and a nitromethylene analogue) for insects versus mammals is not shared by three of their N-unsubstituted imine derivatives or by nicotine or epibatidine. The same selectivity pattern is evident at the receptor level, i.e., the insect nicotinic acetylcholine receptor (nAChR) versus mammalian nAChR subtypes (alpha1, alpha3, alpha4, and alpha7) assayed independently. The insect-selective compounds are not protonated with a nitroimine, cyanoimine, or nitromethylene group and the mammalian-selective compounds are ionized at physiological pH. We propose that the negatively charged tip of the nitro or cyano group (not a partial positive charge at imidazolidine N-1 as suggested earlier) interacts with a putative cationic subsite of the insect nAChR. This contrasts with the mammalian nAChRs where the iminium cation (+C-NH2 <--> C =+NH2) of the neonicotinoid imine derivatives or ammonium nitrogen of nicotine or epibatidine interacts with the anionic subsite.

Role of C3 cleavage in monocyte activation during extracorporeal circulation.

BACKGROUND: We previously demonstrated that inhibiting formation of terminal complement components (C5a and C5b-9) prevents platelet and neutrophil (PMN) but not monocyte activation during simulated extracorporeal circulation (SECC). This study examined whether earlier complement inhibition during SECC, blocking C3a formation, would additionally prevent monocyte activation. METHODS AND RESULTS: SECC was established by recirculating heparinized whole blood from human volunteers on a membrane oxygenator. CAB-2, a chimeric protein constructed from genes encoding the complement regulatory proteins CD46 and CD55, inactivates the C3/C5 convertases and blocks in vitro generation of C3a, C5a, and C5b-9. CAB-2 was used in 4 experiments at a final concentration of 300 micrograms/mL and 4 experiments at 30 micrograms/mL; 4 control runs used vehicle alone. Samples were assayed for C3a and C5b-9, monocyte activation (CD11b upregulation), PMN activation (CD11b upregulation and elastase release), and platelet activation (P-selectin expression and monocyte-platelet conjugate formation). CAB-2 at both doses significantly inhibited formation of C3a and C5b-9 during SECC. High-dose CAB-2 significantly blocked monocyte and PMN CD11b upregulation and PMN elastase release. CAB-2 also inhibited formation of platelet activation-dependent monocyte-platelet conjugates. CONCLUSIONS: Blockade of complement activation early in the common pathway inhibited monocyte CD11b upregulation during SECC, suggesting that early complement components contribute most to monocyte activation during SECC. As expected, PMN and platelet activation were blocked by terminal complement inhibition. This investigation further elucidates the relation between complement and blood cell activation during simulated cardiopulmonary bypass.

Intestinal helminth infections amongst school children in the Serian District of Sarawak.

School children from 3 primary and 2 secondary schools in Sarawak were examined for the presence of gastrointestinal helminths. One primary school and 1 secondary school were located in a town (Serian), the other primary and secondary schools were in the countryside outside Serian. The intestinal helminths detected were Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis and hookworm. Children from the rural schools had higher numbers of eggs in their faeces than those from the Serian schools. Children from the rural primary schools had higher number of eggs than those from the rural secondary school. The prevalence of Ascaris, Trichuris and hookworms in male and female and in primary and secondary school children was recorded.

New stability conditions for Hopfield networks in partial simultaneous update mode.

Cernuschi-Frías has proposed a partial simultaneous updating (PSU) mode for Hopfield networks. He also derived sufficient conditions to ensure global stability. In this letter, a counter-example is given to illustrate that the PSU sequence may converge to limited cycles even if one uses a connection matrix satisfying the Cernuschi-Frías conditions. Then, new sufficient conditions ensuring global convergence of a Hopfield network in PSU mode are derived. Compared with the result of fully parallel mode case, the new result permits a little relaxation on the lower bound of the main diagonal elements of the connection matrix.