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Given that reliable markers for early ischemic brain damage are lacking, we set out to test whether pimonidazole can be used as a reliable tool in the quantification of hypoxic insults, at early time points following experimental stroke.We have used semi-quantitative Western blotting detection of pimonidazole adducts in a rat model of reversible middle cerebral artery occlusion (MCAO), treated with remote post-conditioning.First, we demonstrated that a linear relationship exist between pimonidazole binding in the ischemic hemisphere and duration of ischemia, in animals subjected to 5, 15, 30, or 60 minutes of occlusion followed by 120 minutes of reflow. Then we showed a significant reduction in pimonidazole binding in the infarcted hemisphere, when rats with 60 minutes of MCAO, immediately after establishment of cerebral reflow, had 3×15 minutes intermittent hind limb ischemia followed by 24-hour survival. We analysed the middle cerebral arteries from animals with 60 minutes of MCAO and early remote post-conditioning, followed by 30 minutes, 24, or 48 hours of reflow. At 24 hours of reflow increases in phosphorylated protein kinase C-alpha with concomitantly increased levels of p38 phosphorylation were observed.Our investigation demonstrates that pimonidazole can be used for quantifying ischemic impact in stroke, even after very short survival times. It furthermore shows that early remote post-conditioning reduces ischemic damage, probably through hyperpolarization and reduced reflow vasospasm in the conduit middle cerebral arteries.

The objective of this article was to qualify and test the recommendations of a national Danish report. We conducted an investigation on the readmittance rate as well as reasons for readmittance in a patient cohort defined through the process of internal audit at the Emergency Department at Zealand University Hospital, Køge, Denmark.A retrospective, descriptive study of admitted patients in November 2014, including a total of 1,440 patients. Data and parameters were obtained from electronic patient records.A total of 162 patients were readmitted within 30 days from their initial admission (11% of the cohort). Of this group, 139 (86%) readmittances were unpreventable or planned. Readmissions caused by missed diagnosis or insufficient treatment accounted for 8% and 6%, respectively. The median time until readmission in these cases were two and four and a half days, respectively. The median time to readmission for the unpreventable readmissions ranged from 13 to 18.5 days.In terms of patient safety, our data support a seven-day observation period for readmission rates when measuring or monitoring quality of care in emergency departments.none.none.

Monoclonal antibody (mAb) based affinity resins usually suffer from low binding capacity, most probably as a result of steric hindrance by the large 150 kDa size of the mAb and a random immobilisation approach. The present work investigates the influence of a variety of factors on dynamic binding capacity (DBC) such as pore/ligand size ratio, accessibility of ligand and ligand density. The effect of pore/ligand size ratio was investigated using Fab and scFv fragments on various resins with different pore sizes. The accessibility of the ligand was investigated by a site-directed immobilisation approach, where three C-terminal tags, PPKPPK, FLAG™ and Cys, were introduced into the Fab fragments for immobilisation on resins via amino-, carboxyl- and thiol-groups, respectively. The scFv fragments were tagged at the C-terminal only with FLAG™ to enable a straight forward purification procedure, and were immobilised to resins via amino- and carboxyl-groups. The target protein had a molecular weight (MW) of 50 kDa. A 3-fold higher dynamic binding capacity at 100% breakthrough (DBC100%) was observed for Fab wild-type (wt) on CNBr-activated Sepharose 4 FF relative to mAb on same resin at the same ligand density. However, no major difference in DBC100% was observed between Fab wt and scFv immobilised on CNBr-activated Sepharose 4 FF at the same ligand density. Thus, further increase of pore/ligand size ratio from Fab to scFv on a resin with average pore size of 300 A, did not seem to be beneficial. Among the tested tags, only the C-terminal Cys tag proved to site-direct the ligands during immobilisation as it allowed the DBC100% to increase 1.6-fold as compared to Fab wt immobilised via amino-groups on CNBr-activated Sepharose 4 FF and Actigel ALD Superflow at the same ligand density. The influence of ligand density was investigated by selecting immobilised Fab Cys on Sulfhydryl-reactive resin. Increasing ligand density from 0.103 to 0.202 μmol/mL resulted in the same utilisation yield (82–85%), whereas a further increase in ligand density from 0.202 to 0.328 μmol/mL resulted in a 20%-unit decrease in utilisation yield and less steep breakthrough curve, suggesting steric hindrance in the pores of the resin. In addition, site-directed affinity ligands resulted in a more pronounced, sigmoid-shaped breakthrough curve, leading to more efficient use of capacity. The highest DBC100% was obtained for Fab Cys on Sulfhydryl-reactive resin and scFv on Actigel ALD Superflow; 11 mg/mL and 10 mg/mL, respectively, as compared to the DBC100% of 0.8 mg/mL for mAb on CNBr-activated Sepharose 4 FF. Pore/ligand size ratio of 3, which was achieved for Fab ligands on the studied resins, was shown to be feasible for capturing a protein in MW of 50 kDa. Totally, a 13.8-fold improvement in DBC100% was achieved with the Fab-based affinity resin coupled via the C-terminal Cys as compared to the mAb-based affinity resin.

The Grid Friendly™ Appliance (GFA) controller was proposed to autonomously switch off appliances by detecting under-frequency events. In this paper, a new frequency responsive load (FRL) controller is proposed by extending the functionality of the existing GFA controller. The proposed FRL controller can autonomously switch on (or off) end-use loads by detecting over-frequency (or under-frequency) events through local frequency measurement. Using the proposed FRL controller, a hierarchical decentralized control strategy is also developed for engaging the end-use loads to provide primary frequency response. The developed control strategy consists of two decision making layers including supervisory and device layers, and has several desirable features for primary frequency control. The FRL controller at the device layer preserves the autonomous operation of the enduse loads. The coordinator at the supervisory layer coordinates autonomous response to overcome the stability issue associated with high penetration of FRLs. The simulation results illustrate the effectiveness of the developed hierarchical decentralized control strategy in providing primary frequency response with the proposed FRL controller.

The wholesale power markets have undergone substantial change over the past few years to deal with increasing penetrations of variable energy resources. The market design and processes have become more complex because of introduction of new products and services to deal with the variability and intermittency of renewable resources. Additionally, the increasing adoption of distributed energy resources presents new challenges for the power grid operations, requiring additional flexibility to ensure stable, reliable, and resilient grid operations. The current market designs, however, do not explicitly value flexibility from resources. This paper presents a novel unit commitment/economic dispatch problem formulation that explicitly values the flexibility offered by resources. The market design is based on the concept of two-part pricing where resources are first paid to make their capacities available, including the flexibility, in the day-ahead markets and subsequently paid performance payments based on real-time dispatch.

The most formidable challenge in assembling a smart grid is the integration of a high penetration of renewables. Demand response (DR), a largely promising concept, is increasingly discussed as a means to cope with the intermittent and uncertain renewables. In this paper, we propose a dynamic market mechanism (DMM) that reaches the market equilibrium through continuous negotiations between key market players. In addition to incorporating renewables, this market mechanism also incorporates a quantitative taxonomy of DR devices, based on the inherent magnitude, runtime, and integral constraints of demands. The DMM is evaluated on an IEEE 118 bus system, a high-fidelity simulation model of the midwestern U.S. power grid. The results show how the proposed mechanism can be utilized to determine combinations of DR devices in the presence of intermittent and uncertain renewables with varying levels of penetration so as to result in a desired level of social welfare.

Mutations in the gene for desmoplakin (DSP) may cause arrhythmogenic right ventricular cardiomyopathy (ARVC) and Carvajal syndrome (CS). Desmoplakin is part of all desmosomes, which are abundantly expressed in both myocardial and epidermal tissue and serve as intercellular mechanical junctions. This study aimed to investigate protein expression in myocardial and epidermal tissue of ARVC and CS patients carrying DSP mutations in order to elucidate potential molecular disease mechanisms. Genetic investigations identified three ARVC patients carrying different heterozygous DSP mutations in addition to a homozygous DSP mutation in a CS patient. The protein expression of DSP in mutation carriers was evaluated in biopsies from myocardial and epidermal tissue by immunohistochemistry. Keratinocyte cultures were established from skin biopsies of mutation carriers and characterized by reverse transcriptase polymerase chain reaction, western blotting, and protein mass spectrometry. The results showed that the mutation carriers had abnormal DSP expression in both myocardial and epidermal tissue. The investigations revealed that the disease mechanisms varied accordingly to the specific types of DSP mutation identified and included haploinsufficiency, dominant-negative effects, or a combination hereof. Furthermore, the results suggest that the keratinocytes cultured from patients are a valuable and easily accessible resource to elucidate the effects of desmosomal gene mutations in humans.

The inflammatory response plays a pivotal role in propagating injury of intracerebral hemorrhage (ICH). Glucagon-like-peptide-1 (GLP-1) is a hormone with antidiabetic effect and may also have antiinflammatory properties. Despite consensus that the glucoregulatory action is mediated by the GLP-1 receptor (GLP-1R), mechanisms in the brain remain unclear. We investigated the effect of a long-acting GLP-1 analog, liraglutide, and its truncated metabolite, GLP-1(9–36)a from dipeptidyl peptidase-4 (DPP-4) cleavage in ICH-induced brain injury. Primary outcomes were cerebral edema formation, neurobehavior, and inflammatory parameters. GLP-1(9–36)a, GLP-1R inhibitor, adenosine monophosphate-activated protein kinase (AMPK) phosphorylation inhibitor and DPP-4 inhibitor were administered to examine the mechanisms of action. Liraglutide suppressed neuroinflammation, prevented brain edema and neurologic deficit following ICH, which were partially reversed by GLP-1R inhibitor and AMPK phosphorylation inhibitor. Liraglutide-mediated AMPK phosphorylation was unaffected by GLP-1R inhibitor, and was found to be induced by GLP-1(9–36)a. GLP-1(9–36)a showed salutary effects on primary outcomes that were reversed by AMPK phosphorylation inhibitor but not by GLP-1R inhibitor. Liraglutide and DPP-4 inhibitor co-administration reversed liraglutide-mediated AMPK phosphorylation and antiinflammatory effects. Liraglutide exerted duals actions and the antiinflammatory effects are partially mediated by its metabolite in a phosphorylated AMPK-dependent manner. Therapies that inhibit GLP-1 degradation may weaken the metabolite-mediated effects.

Aneurysmal rebleeding poses a serious risk in patients with subarachnoid hemorrhage (SAH). Studies have shown that antifibrinolytic therapy with tranexamic acid has a dramatic effect on the rate of rebleeding. Therefore, changes in the fibrinolytic system could be hypothesized. We have used an experimental SAH rat model to demonstrate serial changes in the haemostatic system as evaluated by Thromboelastography® (TEG). In the SAH group, a shorter reaction time (R-time) and higher maximum amplitude (MA) were observed. In the saline group, only a shorter R-time was observed. The study has shown that a hypercoagulable state is present immediately after experimental SAH is induced as determined by TEG. The reduction in R-time and rise in MA observed in the SAH group indicate that blood in the subarachnoid space is necessary to accomplish a full systemic coagulation response. This abnormality in coagulation profile seems to be a response to the acute traumatic event caused by induction of SAH.

Whole body hematocrit/large vessel hematocrit (WBH/LVH ratio) has been examined in 21 normotensive and 35 hypertensive subjects and found to be the same, presumably as an expression of a uniform distribution of the blood volume in the two groups. Moderate renal impairment, the degree of the retinopathy, or the height of the diastolic blood pressure do not seem to influence the ratio figures. The WBH/LVH ratio was examined during treatment with various hypotensive drugs. A significant increase in the ratio was demonstrated during the influence of a single effective intravenous dose of pentolinium. Similar results were obtained during short-term treatment with guanethidine alone and during long-term treatment with guanethidine and hydrochlorothiazide. Treatment with hydrochlorothiazide alone caused an increase in ratio after one week and after three months, but these increases were not significant. The possible causes of these alterations in the WBH/LVH ratio are discussed. The advantage of employing a double isotope technique in preference to a quicker single isotope technique has been analyzed in relation to the determination of the total blood volume in patients during treatment with hypotensive drugs.

Short-term exercise produced a decrease in plasma volume and a rise in B.P. in normotensives and untreated hypertensive patients. The decrease in plasma volume is presumably due to an increased intra-capillary pressure in the contracting muscles. In guanethidine-treated patients there was no change in plasma volume during exercise, probably because of vasodilatation and decreased intra-capillary pressure in the contracting muscles. In several of the guanethidine-treated patients B.P. fell during and after exercise, in some cases so markedly that symptoms of cerebral ischemia developed.

Eleven patients with essential hypertension were treated with hydrochlorothiazide/KCl for three months. A significant fall in total blood volume, exchangeable sodium and blood pressure could be demonstrated both after one to two weeks and after three months of treatment, while a fall in serum sodium and body weight was only significant after one to two weeks of treatment. Thus, hypovolemia may, perhaps, be the explanation for the antipressor effect of thiazide even after three months of treatment.

Thirty-one multiple sclerosis patients in a stable phase were given intensive symptomatic therapy. Isometric and isokinetic measurements of voluntary power were used to assess the effect of the treatment. Twenty-one of the patients were improved, six unchanged and four aggravated. The relative improvement was on the average 35%, best in the weakest patients. Isokinetic measurements revealed a particularly good progress for the fastest movements, probably reflecting reduction of spasticity. The optimum effect of treatment was found on an average after three weeks. Such tests seem to be useful in following and analysing the training and possibly in establishing treatment models.

Coagulation factor VIII (FVIII) is one of the most complex biopharmaceuticals due to the large size, poor protein stability and extensive post-translational modifications. As a consequence, efficient production of FVIII in mammalian cells poses a major challenge, with typical yields two to three orders of magnitude lower than for antibodies. In the present study we investigated CHO DXB11 cells transfected with a plasmid encoding human coagulation factor VIII. Single cell clones were isolated from the pool of transfectants and a panel of 14 clones representing a dynamic range of FVIII productivities was selected for RNA sequencing analysis. The analysis showed distinct differences in F8 RNA composition between the clones. The exogenous F8-dhfr transcript was found to make up the most abundant transcript in the present clones. No correlation was seen between F8 mRNA levels and the measured FVIII productivity. It was found that three MTX resistant, nonproducing clones had different truncations of the F8 transcripts. We find that by using deep sequencing, in contrast to microarray technology, for determining the transcriptome from CHO transfectants, we are able to accurately deduce the mature mRNA composition of the transgene and identify significant truncations that would probably otherwise have remained undetected.

Object The authors investigated early changes in the cerebral arteries of rats that occur after subarachnoid hemorrhage (SAH). Methods Messenger RNA was investigated by performing microarray and quantitative real-time polymerase chain reaction (PCR) analyses, and protein expression was shown by performing immunohistochemical studies. The array data indicated that the initial processes that occur after SAH involve activation of genes involved in angiogenesis, inflammation, and extracellular matrix (ECM) remodeling. The real-time PCR investigation confirmed upregulation of genes that were observed using the microarray to be regulated, including iNOS, MMP13, and cxcl2. The authors also verified the upregulation of previously implicated genes for G-protein–coupled receptors (endothelin B [ETB], angiotensin 1 [AT1], and AT2) and metalloproteinase 9. The results of an immunohistochemical study confirmed that receptor genes that were seen to be regulated produced an increase in protein expression. Double immunostaining of rat cerebral arteries with endothelial cell– or smooth-muscle cell–specific antibodies verified that an increase in ETB, 5-hydrotryptamine (5-HT1B), and 5-HT1D receptor expression occurs in smooth-muscle cells. Conclusions Processes occurring after SAH lead to enhanced arterial contractility and ECM remodeling either directly or through angiogenesis and inflammation. These processes are active via an increase in metalloproteinase expression, the presence of proangiogenic factors, and the expression of proinflammatory genes.

The pathogenesis of cerebral ischaemia after subarachnoid haemorrhage (SAH) still remains elusive. The purpose of the present study was to examine whether specific protein kinas C (PKC) inhibition in rats could alter the transcriptional SAH induced Endothelin (ET) type B and 5-hydroxytryptamine type 1B (5-HT1B) receptor upregulation and prevent the associated cerebral blood flow (CBF) reduction. The PKC inhibitor RO-31-7549 or vehicle was injected intracisternally after the induced SAH in rats ( n = 3 to 10 in each groups for each method). The involvement of the PKC isoforms was investigated with Western blot; only PKCδ and PKCα subtypes were increased after SAH RO-31-7549 treatment abolished this. At 2 days after the SAH basilar and middle cerebral arteries were harvested and the contractile response to endothelin-1 (ET-1; ETA and ETB receptor agonist) and 5-carboxamidotryptamine (5-CT; 5-HT1 receptor agonist) were investigated with a myograph. The contractile responses to ET-1 and 5-CT were increased ( P < 0.05) after SAH compared with sham operated rats. In parallel, the ETB and 5-HT1B receptor mRNA and protein expression were significantly elevated after SAH, as analysed by quantitative real-time polymerase chain reaction and immunohistochemistry, respectively. Administration of RO-31-7549 prevented the upregulated contraction elicited by application of ET-1 and 5-CT in cerebral arteries and kept the ETB and 5-HT1B receptor mRNA and protein levels at pre-SAH levels. Regional and global CBF evaluated by an autoradiographic technique were reduced by 60%±4% after SAH ( P < 0.05) and prevented by treatment with rO-31-7549. Our study suggests that PKC plays an important role in the pathogenesis of cerebral ischaemia after SAH.

Conscription has been employed for more than a century. To evaluate the course and fate of conscripts deemed mentally unfit for immediate continued service, a prospective questionnaire study of conscripts referred for mental evaluation was conducted. Questionnaires were completed at the time of enrollment, time of admittance for evaluation, time of discharge from their evaluation and at 1-year follow-up. Out of a class of 6949 conscripts, 93 were referred for psychiatric evaluation and 69 of those referred decided to enter the study (60 of these were in time deemed unfit for further military service). At the time of admittance, on average after 79 days of service, there was a significant 36%, 34%, 20% and 20% increase in depression, sleep, somatization and interpersonality scores, respectively. At the time of discharge 14 days after admittance, the symptom scores had dropped to a level comparable to scores at the time of enrollment. At 1-year follow-up, the descending trend in scores was maintained, though not reaching statistical significance compared with scores at time of discharge. The scores at 1-year follow-up were also, except for a higher depression score, comparable with the scores of a control group of conscripts evaluated 1 year after service. The psychological profile of conscripts referred for evaluation was different at the time of enrollment compared with conscripts in general, the scores being between 9% (interpersonality) and 29% (depression) higher. They also had a higher prevalence of adverse family events in their history. The course observed is indifferent from other situations of enforced stress.

The cellular mechanisms responsible for cerebral vasospasm (CVS) occurring after subarachnoid hemorrhage (SAH) have been of major interest over the past 50 years. The present review describes how each of the discrete anatomic components that comprise the cerebral artery may contribute to the pathology of CVS. The blood extravasated after SAH is hemolyzed and undergoes degradation with resultant production of free radicals, known to be powerful initiators of vascular damage. An inflammatory response is generated activating both leukocytes and platelets with subsequent release of inflammatory agents. The cerebral artery affected by CVS undergoes phenotypic change involving both the endothelial and smooth muscle cells. In the endothelium the production of nitric oxide and prostacyclin is affected. In the smooth muscle cells signal transduction pathways that enhance the function of the contractile proteins and induce the upregulation of contractile receptors are activated. In parallel, there is evidence that nervous reflex pathways involving the trigeminal ganglion and the hypothalamus are activated. However, the relative contributions of each of the systems are speculative. Therapy may be directed at disrupting the cascade leading from the SAH insult to CVS or at overcoming the dysfunction incurred by CVS; possible therapeutical interventions are considered.

Overall the aim has been to characterise the in vitro upregulation of the 5-hydroxytryptamine 1B/1D (5-HT(1B/1D)) and endothelin (ET) receptors in the cerebral circulation as well as demonstrating phenotypic changes in a pathophysiological model mimicking subarachnoid haemorrhage. The first part describes receptor upregulation using organ culture of whole vessel segments from both rat and man. The upregulation is dependent on gene transcription and translation; thus, an active signalling pathway is involved. Further analysis of this pathway revealed that protein kinase C is especially important in the upregulation process of the ET(B) receptor. The second part attempts to take the concept of receptor upregulation into the pathophysiological realm, especially focusing on experimental subarachnoid haemorrhage. Changes similar to those observed using the in vitro organ culture technique were observed in vivo. Especially the ET(B) receptor upregulates and perhaps more importantly, this appears to be associated with a generally increased sensitivity towards ET-1. Now, even at physiological concentrations of ET-1 were appreciable contraction observed. Similar results were obtained when studying the upregulation of the 5-HT(1B) receptor; the upregulation imparts increased sensitivity of the cerebral arteries towards serotonin. The increased functional responses represent a significant contribution towards the understanding of cerebral vasospasm after subarachnoid haemorrhage. Finally, it is revealed that the ET(B) receptor is more frequently present in human arteries from patients with cerebrovascular disease thus substantiating the idea of receptor upregulation as part of human cerebrovascular pathology.

The vasomotor effects of triptans in the middle cerebral artery (MCA) of rats were studied using the pressurised arteriography method and in vitro vessel baths. Using the arteriograph, MCAs from Sprague-Dawley rats were mounted on two glass micropipettes, pressurised to 85 mm Hg and luminally perfused. Luminally added 5- hydroxytryptamine (5-HT), sumatriptan and rizatriptan induced maximal dilatations of 22 ± 4, 10 ± 2 and 13 ± 5%, respectively, compared to the resting diameter. The relaxant effect of sumatriptan was blocked by the 5- HT1B/1D receptor selective antagonist GR 55562 (10–6M). The use of Nω-nitro-L-arginine and charybdotoxin revealed that the dilatation involved both nitric oxide and endothelially derived hyperpolarising factor. Thus, the earlier demonstrated expression of 5-HT1B/1D immunoreactivity in the endothelium may well translate into a relaxant response to 5-HT and triptans. Using the vessel bath technique, MCA segments were mounted on two metal wires. The relaxant responses to sumatriptan could not be reproduced using this model; instead, weak contractile responses (6 ± 3% of submaximal contractile capacity) were observed. The difference in observations between the experimental models may be related to the maintenance of shear stress in the arteriograph.

1 Organ culture has been shown to upregulate both endothelin (ET) and 5-hydroxytryptamine 1B/1D (5-HT(1B/1D)) receptors in rat cerebral arteries. The purpose of the present study was to investigate the involvement of protein kinases, especially protein kinases C (PKC) and A (PKA) in this process. 2 The effect of inhibiting protein kinases during organ culture with staurosporine (unspecific protein kinase inhitor), RO 31-7549 (specific inhibitor of classical PKC's) and H 89 (specific inhibitor of PKA) was examined using in vitro pharmacological examination of cultured vessel segments with ET-1 (unspecific ET(A) and ET(B) agonist), S6c (specific ET(B) agonist) and 5-CT (5-HT(1) agonist). Levels of mRNA coding for the ET(A), ET(B), 5-HT(1B) and 5-HT(1D) receptors were analysed using real-time RT-PCR. 3 Classical PKC's are critically involved in the appearance of the ET(B) receptor; co-culture with RO 31-7549 abolished the contractile response (6.9+/-1.8%) and reduced the ET(B) receptor mRNA by 44+/-4% as compared to the cultured control. Correlation between decreased ET(B) receptor mRNA and abolished contractile function indicates upstream involvement of PKC. 4 Inhibition of PKA generally had an enhancing effect on the induced changes giving rise to a 7-25% increase in E(max) in response to ET-1, S6c and 5-CT as compared to the cultured control. 5 Staurosporine inhibited the culture induced upregulation of the response of both the ET(A) and the 5-HT(1B/1D) receptors, but had no significant effect on the mRNA levels of these receptors. This lack of correlation indicates an additional downstream involvement of protein kinases. British Journal of Pharmacology (2002) 137, 118-126. doi:10.1038/sj.bjp.0704838 (Less)

Summary: This pilot study looks at the frequency of suicide among Danish soldiers who took part in the UN mandated forces (UNMF) during the 1990's. In a contingent of nearly 4000 Danish UN soldiers four suicides were documented, two of whom committed suicide less than one month before deployment and two who committed suicide within a year after discharge from mission. Contributing factors, prevention strategies, and implications for future research are discussed.

In recent decades, moves toward higher integration of Renewable Energy Resources have called for fundamental changes in both the planning and operation of the overall power grid. One such change is the incorporation of Demand Response (DR), the process by which consumers can adjust their demand in a flexible manner. This paper presents a survey of various aspects of DR including the different types of participants, as well as the underlying challenges and the overall potential of DR when it comes to large-scale implementations. Benefits of DR as reported in the literature for performance metrics such as frequency control and price control, as well as methods for ensuring privacy are discussed. A quantitative taxonomy of DR recently proposed in the literature based on the inherent magnitude, run-time, and integral constraints is discussed and its integration with economic dispatch is explored.

The revised Scandinavian Neurotrauma Committee (SNC) guidelines on management of patients with head trauma include an option for measurement of S100B in peripheral blood with 100% sensitivity for neurosurgical intervention. A medical technology assessment was conducted to evaluate any impact of using S100B on the use of computed tomographies (CT) of the brain and admission for observation.Patients referred for assessment of head injury over a period of 1.5 months had their blood sampled for measurement of S100B in serum. Results were not available to the treating physician and treatment was conducted according to existing practice. Patient records were reviewed retrospectively and post hoc divided into two groups depending on whether the SNC criteria for taking the blood sample were met. The use of CT and admission was analysed.A total of 39 patients had their blood sampled for analysis. In all, 12 patients were excluded in pursuance of SNC guidelines, which left 27 patients for analysis. A total of 15 patients had abnormally high S100B levels. Using the SNC criteria, only eight of these qualified a priori for blood sampling. Furthermore, seven of the 11 patients who were admitted had normal S100B levels.The number of patients with an above-threshold concentration of S100B was almost equally distributed between those fulfilling the SNC criteria for S100B assessment and those who could have been discharged without further evaluation. Using S100B as a screening tool may lead to an increase in the use of CTs of the brain. In relation to admission, measurement of S100B may contribute to the adoption of an appropriate observation strategy.not relevant.not relevant.

In this paper, we propose a dynamic market mechanism that converges to the desired market equilibrium. Both locational marginal prices and the schedules for generation and consumption are determined through a negotiation process between the key market players. In addition to incorporating renewables, this mechanism accommodates both consumers with a shiftable Demand Response and an adjustable Demand Response. The overall market mechanism is evaluated in a Day Ahead Market and is shown in a numerical example to result in a reduction of the cost of electricity for the consumer, as well as an increase in the Social Welfare.

5-hydroxytryptamine (5-HT) is thought to be involved in migraine headache and the pathophysiology of cerebrovascular diseases. Previous data show that organ culture induces a phenotypic change in cerebral vessels. Therefore we investigated if these changes also applied for the vasoconstrictive 5-HT receptors. Rat cerebral arteries express 5-HT 2 receptors. Using organ culture we observed a phenotypic change with a selective up-regulation of 5-HT IB/ID receptors. This was revealed by an increased sensitivity to the selective 5-HT IB/ID agonist 5-CT after organ culture (pEC 50(fresh) 5.6±0.2 and pEC 50(cultured) 6.8 ± 0.4). The response was inhibited by the 5-HT IB/ID selective antagonist GR55562 (pEC 50(fresh) 5.1 ± 0.2 and pEC 50(cultured) 6.0 ± 0.3). The organ model might mimic the phenotypic changes during cerebrovascular diseases.

An unusual case of very-late-onset metachromatic leukodystrophy (MLD) with dementia was studied. The patient was a 41-year-old male who presented with mild dementia and a single generalized tonic clonic seizure. Neuropsychological assessment demonstrated mild amnesia, visuospatial dysfunction and attention deficits with a slow psychomotor speed. MR brain imaging displayed confluent hyperintensities of periventricular and subcortical white matter. Low levels of arylsulfatase A confirmed the diagnosis. Impaired cortical glucose metabolism especially of the medial temporal and frontal cortices was observed using positron emission tomography and fluor-18-labeled fluorodesoxyglucose. The neuropsychological deficits are related to the location of deficits in glucose metabolism.

Udgivelsesdato: 2000-Oct-27 The low density lipoprotein (LDL) receptor is responsible for removing the majority of the LDL cholesterol from the plasma. Mutations in the LDL receptor gene cause the disease familial hypercholesterolemia (FH). Approximately 50% of the mutations in the LDL receptor gene in patients with FH lead to receptor proteins that are retained in the endoplasmic reticulum (ER). Misfolding of mutant LDL receptors is a probable cause of this ER retention, resulting in no functional LDL receptors at the cell surface. However, the specific factors and mechanisms responsible for retention of mutant LDL receptors are unknown. In the present study we show that the molecular chaperone Grp78/BiP co-immunoprecipitates with both the wild type and two different mutant (W556S and C646Y) LDL receptors in lysates obtained from human liver cells overexpressing wild type or mutant LDL receptors. A pulse-chase study shows that the interaction between the wild type LDL receptor and Grp78 is no longer detectable after 2(1/2) h, whereas it persists for more than 4 h with the mutant receptors. Furthermore, about five times more Grp78 is co-immunoprecipitated with the mutant receptors than with the wild type receptor suggesting that Grp78 is involved in retention of mutant LDL receptors in the ER. Overexpression of Grp78 causes no major alterations on the steady state level of active LDL receptors at the cell surface. However, overexpression of Grp78 decreases the processing rate of newly synthesized wild type LDL receptors. This indicates that the Grp78 interaction is a rate-limiting step in the maturation of the wild type LDL receptor and that Grp78 may be an important factor in the quality control of newly synthesized LDL receptors.

Denmark has been engaged in the Afghanistan war and as a result, Rigshospitalet has received a number of multi-traumatized Danish soldiers. Lesions sustained in armed conflict differ from their civilian counterparts and knowledge of the pathophysiology related to these types of lesions is essential when engaging in the intensive care of these patients.The study was conducted as a retrospective survey of Danish soldiers evacuated from Afghanistan to the Intensive Care Unit at Rigshospitalet in the 2002-2012 period. The following data were recorded: age, gender, hospitalization (days), mortality, organ involvement, respiratory therapy, dialysis, circulatory supportive care, antibiotic treatment and bacteriology. Furthermore, Acute Physiology and Chronic Health Evaluation, Simplified Acute Physiology Score and Sequential Organ Failure Assessment scores were calculated.A total of twenty patients were identified and included in the study. All patients had sustained serious blast injuries as a result of explosion. Primarily the central nervous system, respiratory, musculoskeletal and abdominal systems were affected by the explosions. Eighteen patients survived to discharge and two patients died.Explosion was the most frequent cause of injury in all cases and caused damage to several organ systems. Infections after combat injuries are a major problem because of the different microbiological profile.The use of explosives has been and remains a substantial part of warfare, and this review has showed us that the knowledge of the mechanism of injury is indeed essential, and that intelligence on the microbiological flora of the geographical location of the conflict is essential.not relevant.not relevant.

Background Criteria for Systemic Inflammatory Response Syndrome (SIRS) are recognized as operational criteria for detecting a possible bacteriaemia. Given the sensitivity and specificity of the criteria, clinical judgment is crucial in detecting the condition. The aim of the study was retrospectively to identify clinical characteristics of patients not primarily thought of as having bacteriaemia, yet harboring the condition.

Objectives: Hypothermia is still unproven as beneficial treatment in human stroke, although in animal models, conditioning the brain with hypothermia has induced tolerance to insults. Here, we delineate the feasibility of drug-induced mild hypothermia in reducing ischemic brain damage when conditioning before (preconditioning) and after (postconditioning) experimental stroke. Methods: Hypothermia was induced in rats with a bolus of 6 mg/kg talipexole followed by 20 h continuous talipexole infusion of 6 mg/kg in total. Controls received similar treatment with saline. The core body temperature was continuously monitored. In preconditioning, hypothermia was terminated before either reversible occlusion of the middle cerebral artery (MCAO) for 60 min or global ischemia for 10 min with 2-vessel occlusion and hypotension. In postconditioning, rats experienced 60 min of MCAO before hypothermia was induced either immediately or with 3 h delay. Rats survived ischemia for 2, 7 or 90 days. Infarct volumes were quantified by stereology. Additional experiments of methodological relevance were included in the study. Results: Talipexole induced mild hypothermia (35.1 ± 1.1 to 36.0 ± 0.5°C) for Conclusion: Talipexole is registered for use as a dopamine substitute in humans with Parkinson's disease. Although dosages cannot be directly translated to patients, our study exemplifies in an animal model that drug-induced hypothermia in a clinical setting might reduce cerebral ischemic damage before neuro- and cardiac surgical procedures and after stroke.

Intraventricular hemorrhage (IVH) subsequent to intracerebral hemorrhage (ICH) or subarachnoid hemorrhage (SAH) is associated with high mortality and morbidity. The use of fibrinolytic agents to treat this condition has previously been reported in small clinical trials with limited numbers of patients. Variability regarding inclusion criteria, method of administration and outcome have made it difficult to draw firm conclusions regarding the efficacy of antifibrinolytic therapy. Nine patients with CT-diagnosed IVH were treated with Alteplase intrathecally for 3 to 5 days according to the CT-verified clearance of IVH. After the treatment period, a repeat CT scan was performed to evaluate treatment effect.

An update of published clinical advances in the treatment of cerebral vasospasm after subarachnoid haemorrhage was provided. Searching MEDLINE using the search terms "cerebral vasospasm" and "clinical trials" 46 papers were identified that had been published since the International Conference on Cerebral Vasospasm in Istanbul, Turkey in 2006. Of these 26 were either safety studies or case reports leaving 20 papers for consideration. The major topics covered were calcium antagonists, magnesium sulphate, statins, and fasudil hydrochloride. The studies published did not reach an impact justified recommended routine use, but certainly as options. Results of the CONSCIOUS trials on endothelin receptor antagonists are awaited.

Canine coagulation factor VII (FVII) deficiency can be hereditary or acquired and may cause life threatening bleeding episodes if untreated. FVII procoagulant activity can be measured by FVII activity (FVII:C), but assays for measurement of canine specific FVII antigen (FVII:Ag) have not been available to date. In this study, a canine specific ELISA for measurement of FVII:Ag in plasma was developed and validated. The FVII:Ag ELISA correctly diagnosed homozygous and heterozygous hereditary FVII deficiency. Together with activity based assays, such as FVII:C, the FVII:Ag ELISA should be valuable in the diagnosis of hereditary canine FVII deficiency.

Objective: Delayed cerebral vasospasm has long been recognized as an important cause of poor outcome after an otherwise successful treatment of a ruptured intracranial aneurysm, but it remains a pathophysiological enigma despite intensive research for more than half a century. Method: Summarized in this review are highlights of research from North America, Europe and Asia reflecting recent advances in the understanding of delayed ischemic deficit. Result: It will focus on current accepted mechanisms and on new frontiers in vasospasm research. Conclusion: A key issue is the recognition of events other than arterial narrowing such as early brain injury and cortical spreading depression and of their contribution to overall mortality and morbidity.

Thirteen pregnancies in ten women on oxcarbazepine (OXC) monotherapy and one pregnancy in a woman on OXC and topiramate therapy were retrospectively analyzed. A significant decrease of ratio plasma concentration of 10-monohydroxy derivate (MHD) of oxcarbazepine to dosage was found by 26.2% during first trimester, by 36.5% during second trimester and by 38.2% during third trimester when compared to pre-pregnant levels. Eight patients experienced seizure deterioration during the pregnancy, five of which had been seizure-free before the pregnancy. In seven pregnancies (50%) the seizure frequency was at least doubled during pregnancy compared to a pre-pregnancy baseline. There was a trend toward a correlation between seizure deterioration and decrease in plasma concentration of MHD.

Because of the concern about environmental chemicals with oestrogenic and anti-oestrogenic effects, there is a need to construct biosensors for classifying such chemicals according to their effect on oestrogen receptor conformation. The conformation of the ligand-binding domains (LBD) of oestrogen receptor-alpha and -beta determine their transcription regulation activity. Some ligands, i.e., the natural oestrogen oestradiol, induce an active conformation allowing interaction with co-activators. In contrast, antagonists like ICI 182, 780, because of their bulky side chains, do not allow an alpha-helix 12 positioning compatible with co-activator binding. Another type of oestrogen receptor-ligand interactions, termed "passive antagonism", was first defined by X-ray crystal structure analysis of receptors in complex with the side chain-less 5,11-cis-diethyl-5,6,11,12-tetrahydrochrysene-2,8-diol (THC). We have now used the ability of peptides selected from phage-displayed peptide libraries to bind conformation specifically to oestrogen receptor-alpha and -beta LBDs to analyse conformations induced by THC and a group of chlorinated biphenyls and their aryl-hydroxylated metabolites, suspected of being environmental chemical disruptors. In oestrogen receptor-beta, THC defined a "passive antagonist" peptide recognition pattern, which was also induced by several antagonistic hydroxylated biphenyls, while a clearly different peptide recognition pattern was induced by their chlorinated agonistic counterparts. In oestrogen receptor-alpha, THC induced a conformation similar to that induced by oestriol and other oestrogen receptor-alpha agonists, which, as evaluated by site-directed mutagenesis, have a functionally important interaction with oestrogen receptor-alpha residue His524. We conclude that the peptide recognition pattern can be used to classify suspected environmental endocrine disruptors according the oestrogen receptor-alpha and -beta conformations they induce.

Three different conformations of the ligand binding domain of the human estrogen receptor (ER-LBD) are observed for the native state when binding an agonist and when binding an antagonist. By conjugating ER-LBD conformation specific peptides to CdS nanoparticles, the three different states can be identified by anodic stripping voltammetry. This electrochemical sensor can detect and distinguish the binding of different ligands to the human estrogen receptor.

Cerebral vasospasm is an important syndrome that afflicts 30% of patients in the aftermath of, and secondary to, subarachnoid hemorrhage. Starting approximately one week after the hemorrhage, the condition worsens the prognosis of the hemorrhage significantly. Apart from general supportive care, no treatment exists for cerebral vasospasm. During the past 50 years, it was thought that the ischemia that signifies poor outcome is more or less exclusively caused by arterial narrowing. However, this idea has recently been challenged by the failure of the drug clazosentan to improve patient outcome, despite reversing vasoconstriction. In this article, we discuss the opinion that factors other than vasoconstriction are important in the pathophysiology and prognosis of cerebral vasospasm. Such factors include global ischemia, disruption of the blood-brain barrier, activation of apoptotic and inflammatory pathways, and cortical spreading depression.

Udgivelsesdato: 2007-null Molecular chaperones assist protein folding, and variations in their encoding genes may be disease-causing in themselves or influence the phenotypic expression of disease-associated or susceptibility-conferring variations in many different genes. We have screened three candidate patient groups for variations in the HSPD1 and HSPE1 genes encoding the mitochondrial Hsp60/Hsp10 chaperone complex: two patients with multiple mitochondrial enzyme deficiency, 61 sudden infant death syndrome cases (MIM: #272120), and 60 patients presenting with ethylmalonic aciduria carrying non-synonymous susceptibility variations in the ACADS gene (MIM: *606885 and #201470). Besides previously reported variations we detected six novel variations: two in the bidirectional promoter region, and one synonymous and three non-synonymous variations in the HSPD1 coding region. One of the non-synonymous variations was polymorphic in patient and control samples, and the rare variations were each only found in single patients and absent in 100 control chromosomes. Functional investigation of the effects of the variations in the promoter region and the non-synonymous variations in the coding region indicated that none of them had a significant impact. Taken together, our data argue against the notion that the chaperonin genes play a major role in the investigated diseases. However, the described variations may represent genetic modifiers with subtle effects.

A new electrochemical DNA sensor providing detection capabilities down to 100 attomol of target DNA has been developed. The method applies CdS, ZnS, and PbS nanoparticles conjugated with short DNA sequences which are immobilized via hybridization with complementary sequences on a gold surface. When the DNA target is added, it can be identified by ousting the existing hybridization between one of the DNA−nanoparticle conjugates and the surface DNA. The nanoparticles remaining at the surface are detected by stripping voltammetry. The setup is constructed to give a signal-off response with a build-in control signal as only one of two different metal sulfide signaling probes on the surface is removed by hybridization with the DNA target. The competition assay is, in principle, label-free since no labels are required for detection after addition of DNA target. The dissociation of PbS nanoparticles from the surface after addition of the DNA target has been imaged by fluid phase AFM.

Upregulation of endothelin B (ETB) and 5-hydroxytryptamine 1B (5-HT1B) receptors via transcription has been found after experimental subarachnoid hemorrhage (SAH), and this is associated with enhanced phosphorylation of the mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase (ERK1/2). In the present study, we hypothesized that inhibition of ERK1/2 alters the ETB and 5-HT1B receptor upregulation and at the same time prevents the sustained cerebral blood flow (CBF) reduction associated with SAH. The ERK1/2 inhibitor SB386023-b was injected intracisternally in conjunction with and after the induced SAH in rats. At 2 days after the SAH, cerebral arteries were harvested for quantitative real-time polymerase chain reaction, immunohistochemistry and analysis of contractile responses to endothelin-1 (ET-1; ETA and ETB receptor agonist) and 5-carboxamidotryptamine (5-CT; 5-HT1 receptor agonist) in a sensitive myograph. To investigate if ERK1/2 inhibition had an influence on the local and global CBF after SAH, an autoradiographic technique was used. At 48 h after induced SAH, global and regional CBF were reduced by 50%. This reduction was prevented by treatment with SB386023-b. The ERK1/2 inhibition also decreased the maximum contraction elicited by application of ET-1 and 5-CT in cerebral arteries compared with SAH. In parallel, ERK1/2 inhibition downregulated ETB and 5-HT1B receptor messenger ribonucleic acid and protein levels compared with the SAH. Cerebral ischemia after SAH involves vasoconstriction and subsequent reduction in the CBF. The results suggest that ERK1/2 inhibition might be a potential treatment for the prevention of cerebral vasospasm and ischemia associated with SAH.

Duchenne's muscular dystrophy (DMD) is a fatal neuromuscular disease caused by absence of dystrophin. Utrophin is a chromosome 6-encoded dystrophin-related protein (DRP), sharing functional motifs with dystrophin. Utrophin's ability to compensate for dystrophin during development and when transgenically overexpressed has provided an important impetus for identifying activators of utrophin expression. The utrophin promoter A is transcriptionally regulated in part by heregulin-mediated, extracellular signal-related kinase-dependent activation of the GABP α/β transcription factor complex. Therefore, this pathway offers a potential mechanism to modulate utrophin expression in muscle. We tested the ability of heregulin to improve the dystrophic phenotype in the mdx mouse model of DMD. Intraperitoneal injections of a small peptide encoding the epidermal growth factor-like region of heregulin ectodomain for 3 months in vivo resulted in up-regulation of utrophin, a marked improvement in the mechanical properties of muscle as evidenced by resistance to eccentric contraction mediated damage, and a reduction of muscle pathology. The amelioration of dystrophic phenotype by heregulin-mediated utrophin up-regulation offers a pharmacological therapeutic modality and obviates many of the toxicity and delivery issues associated with viral vector-based gene therapy for DMD.

Misalignment and instability after cervical laminectomy, performed to treat spondylotic myelopathy, has been described as possible adverse effects. Forty-six consecutively patients on whom laminectomy had been performed in a 4-year period were identified; 7.7 +/- 0.6 years after operation, 11 patients had died and 27 patients were available for follow-up. Postoperative static subluxation was observed in 26% of the patients with an average slip of 3.7 +/- 3.1 mm; 7% had abnormal intervertebral movement displaying 1-2 mm movement from full flexion to full extension. Seventy-four per cent of the patients showed abnormal spinal curvature as judged from radiographs. However, no correlation with outcome was observed. Seventy-four per cent of the patients thought of the result of the operation as either good or fair; objectively, the best long-term effect of the operation was upon arm function. Although some patients develop postural anomaly, laminectomy remains, in terms of instability, a justifiable procedure in the elderly patient with spondylosis.

Object. Cerebral vasospasm following subarachnoid hemorrhage (SAH) leads to reduced blood flow in the brain. Inspired by organ culture—induced changes in the receptor phenotype of cerebral arteries, the authors investigated possible changes in the 5-hydroxytryptamine (HT) receptor phenotype after experimental SAH. Methods. Experimental SAH was induced in rats by using an autologous prechiasmatic injection of arterial blood. Two days later, the middle cerebral artery (MCA), posterior communicating artery (PCoA), and basilar artery (BA) were harvested and examined functionally with the aid of a sensitive in vitro pharmacological method and molecularly by performing quantitative real-time reverse transcription—polymerase chain reaction (PCR). In the MCA and BA the 5-HT1B receptor was upregulated, as determined through both functional and molecular analysis. In response to selective 5-HT1 receptor agonists both the negative logarithm of the 50% effective concentration was increased (one log unit in the MCA and one half unit in the BA), as was the agonist's potency (increased by 50% in the MCA and doubled in the BA). In addition, the authors found an approximately fourfold increase in the number of copies of messenger RNA coding for the 5-HT1B receptor as determined by quantitative real-time PCR. In the PCoA no upregulation of the 5-HT1B receptor was observed. Conclusions. Changes in the receptor phenotype in favor of contractile receptors may well represent the end stage in a sequence of events leading from SAH to the actual development of cerebral vasospasm. Insight into the mechanism of upregulation may provide new targets for developing specific treatment against cerebral vasospasm.