The chronic pelvic pain syndrome is char


The chronic pelvic pain syndrome is characterized by pelvic pain, voiding symptoms and varying degrees of inflammation within expressed prostatic secretions. We evaluated the chemokines monocyte chemoattractant protein 1 (CCL2) and macrophage inflammatory protein-1 alpha (CCL3) in expressed prostatic secretions to identify marker increases associated with inflammatory (IIIA) and noninflammatory (IIIB) chronic pelvic pain syndrome. In addition, chemokine levels were correlated with clinical pain as determined by the National Institutes of Health chronic prostatitis symptom index.

Materials and Methods: Expressed prostatic secretions were collected by digital rectal examination, and evaluated by enzyme linked immunosorbent assays for monocyte chemoattractant protein 1 and macrophage inflammatory protein-la in 154 patients including controls (13), those with benign prostatic hyperplasia (54), chronic pelvic pain syndrome see more IIIA (37) and IIIB (50). Monocyte chemoattractant protein 1 and macrophage inflammatory protein-la levels were compared between IIIA, IIIB and the control subgroups, and correlated

against the chronic prostatitis symptom index and pain subscore using a Spearman test.

Results: Mean levels of monocyte chemoattractant protein 1 in the control, inflammatory benign prostatic hyperplasia, noninflammatory benign prostatic hyperplasia, inflammatory GSK1120212 purchase chronic pelvic pain syndrome and noninflammatory chronic pelvic pain syndrome were 599.4, 886.0, 1,636.5, 3,261.2 and 2,272.7 pg/ml, respectively. Mean levels of macrophage inflammatory protein-1 alpha in the control, inflammatory benign prostatic hyperplasia, noninflammatory benign prostatic hyperplasia, IIIA chronic pelvic pain syndrome and IIIB chronic pelvic pain syndrome were 140.1, 299.4, 238.7, 1,057.8 and 978.4 pg/ml, respectively. For each cytokine both

chronic pelvic pain syndrome subtypes had statistically higher levels than the control group and patients with benign prostatic hyperplasia (p = 0.0002). selleck kinase inhibitor Receiver operating curves using monocyte chemoattractant protein 1 levels greater than 704 pg/ml and macrophage inflammatory protein-la greater than 146 pg/ml identified patients with chronic pelvic pain syndrome with an accuracy of 90% from control patients. Macrophage inflammatory protein-la levels (p = 0.0007) correlated with the pain subscore of the chronic prostatitis symptom index while monocyte chemoattractant protein 1 (p = 0.71) did not.

Conclusions: Monocyte chemoattractant protein I and macrophage inflammatory protein-la within the prostatic fluid in both chronic pelvic pain syndrome subtypes provide candidate future biomarkers for chronic pelvic pain syndrome. In addition, macrophage inflammatory protein-la increase in expressed prostatic secretions provides a new marker for clinical pain in chronic pelvic pain syndrome patients.

This work has important implications for the understanding and tr

This work has important implications for the understanding and treatment of sexual dysfunction in people including delayed/premature ejaculation, involuntary vaginal spasms, and pain during intercourse. JQ1 concentration Published by Elsevier Ltd on behalf of IBRO.”
“Purpose: Altered sensory processing in interstitial cystitis/painful bladder syndrome cases may result from a deficiency of the central nervous system to adequately filter incoming visceral afferent information. We used prepulse inhibition as an operational measure of sensorimotor gating to examine early pre-attentive stages of information processing in females with interstitial

cystitis/painful bladder syndrome and healthy controls.

Materials and Methods: We assessed prepulse inhibition in 14 female patients with interstitial cystitis/painful bladder syndrome

and 17 healthy controls at 60 and 120-millisecond prepulse-to-startle stimulus intervals. We evaluated group differences in prepulse inhibition, and relationships between prepulse inhibition, neuroticism and acute stress ratings.

Results: Patients showed significantly decreased prepulse inhibition at 60 and 120-millisecond prepulse intervals. The prepulse inhibition deficit was related to acute stress ratings in the patients. However, increased neuroticism appeared to mitigate the prepulse inhibition deficit in those GSK872 with interstitial cystitis/painful bladder syndrome, possibly reflecting greater vigilance.

Conclusions: Compared to healthy controls,

female patients with interstitial cystitis/painful bladder syndrome had decreased ability to adequately filter incoming information and perform appropriate sensorimotor gating. These results suggest that a possible mechanism for altered interoceptive information processing in interstitial cystitis/painful bladder syndrome cases may be a general deficit in filtering mechanisms due to altered pre-attentive processing.”
“In larval lamprey, spinal locomotor activity can be initiated by pharmacological microstimulation from the following higher order brain locomotor areas [Paggett Pyruvate dehydrogenase lipoamide kinase isozyme 1 et al. (2004) Neuroscience 125:25-33; Jackson et al. (2007) J Neurophysiol 97:3229-3241]; rostrolateral rhombencephalon (RLR); ventromedial diencephalon (VMD); or dorsolateral mesencephalon (DLM). In the present study, pharmacological microstimulation with excitatory amino acids (EAAs) or their agonists in the brains of in vitro brain/spinal cord preparations was used to determine the sizes, pharmacology, and organization of these locomotor areas. First, the RLR, DLM and VMD locomotor areas were confined to relatively small areas of the brain, and stimulation as little as 50 mu m outside these areas was ineffective or elicited tonic or uncoordinated motor activity.

At 2 years, niacin therapy had significantly increased the median

At 2 years, niacin therapy had significantly increased the median HDL cholesterol level from 35 mg per

selleck inhibitor deciliter (0.91 mmol per liter) to 42 mg per deciliter (1.08 mmol per liter), lowered the triglyceride level from 164 mg per deciliter (1.85 mmol per liter) to 122 mg per deciliter (1.38 mmol per liter), and lowered the LDL cholesterol level from 74 mg per deciliter (1.91 mmol per liter) to 62 mg per deciliter (1.60 mmol per liter). The primary end point occurred in 282 patients in the niacin group (16.4%) and in 274 patients in the placebo group (16.2%) (hazard ratio, 1.02; 95% confidence interval, 0.87 to 1.21; P = 0.79 by the log-rank test).


Among patients with atherosclerotic cardiovascular disease and LDL cholesterol levels of less than 70 mg per deciliter (1.81 mmol per liter), there was no incremental clinical benefit from the addition of niacin to MK-4827 statin therapy during a 36-month follow-up period, despite significant improvements in HDL cholesterol and triglyceride levels. (Funded by the National Heart, Lung, and Blood Institute and Abbott Laboratories; AIM-HIGH ClinicalTrials.gov number, NCT00120289.)”

Voiding cystourethrogram is an invasive test that evokes anxiety. Our primary aim was to determine whether midazolam is beneficial in decreasing anxiety in children who undergo voiding cystourethrogram. Secondary aims were an examination of parent anxiety, health care professional perceptions and post-procedure behavioral outcomes in children after voiding cystourethrogram.

Materials these and Methods: A total of 44 children were randomized to placebo or oral midazolam before voiding cystourethrogram in double-blind fashion. The Modified Yale Preoperative Anxiety Scale was used to evaluate child behavior before and during voiding cystourethrogram,

and the Post Hospitalization Behavior Questionnaire was used to investigate any short-term and intermediate-term behavioral outcomes. The State-Trait Anxiety Inventory was used to evaluate parent personal anxiety during voiding cystourethrogram. A separate questionnaire was administered to radiology staff. Statistical analysis included the 2-sample t and Fisher exact tests.

Results: There was no difference in Modified Yale Preoperative Anxiety Scale scores in children randomized to midazolam or placebo. There was also no significant difference in parent anxiety. Radiology care providers identified no reliable benefit when blinded to sedation vs placebo. We did not note any post-procedural behavior issues after voiding cystourethrogram at up to 6 months of followup.

Conclusions: Midazolam may not significantly help with child or parent anxiety during voiding cystourethrogram. No reliable benefit was noted according to radiology health care provider perception and there was no significant post-procedural behavior benefit.

4/V787A, Na(v)1 4/V787C, and Na(v)1 4/V787K cDNAs, expressed wild

4/V787A, Na(v)1.4/V787C, and Na(v)1.4/V787K cDNAs, expressed wildtype and mutated channels with the auxiliary

beta 1 subunit in Xenopus oocytes, and used the two-electrode voltage clamp technique to examine the effects of these mutations on channel inhibition by four SCI insecticides (indoxacarb, its bioactivated metabolite DCJW, metaflumizone, and RH3421). Mutations at Val787 affected SCI insecticide sensitivity in a manner that was independent of mutation-induced changes in slow inactivation gating. Sensitivity to inhibition by 10 mu M indoxacarb was significantly increased in all three mutated Go6983 channels, whereas sensitivity to inhibition by 10 mu M metaflumizone was significantly reduced in Na(v)1.4/V787A channels and completely abolished in Na(v)1.4/V787K channels. The effects of Val787 mutations on metaflumizone were correlated with the hydrophobicity of the substituted amino acid rather than

the extent of slow inactivation. None of the mutations at Val787 significantly affected the sensitivity to inhibition by DCJW or RH3421. These results demonstrate that the impact of mutations at Val787 on sodium channel inhibition by SCI insecticides depend on the specific insecticide examined and is independent of mutation-induced changes in slow inactivation gating. We propose that Val787 may be a unique determinant of metaflumizone binding. (C) 2012 Elsevier Inc. All rights reserved.”
“Basic science literature ABT-737 abounds with molecules that promise to ameliorate almost any disease, from curing cancer to slowing the aging process itself. However, most of these compounds will never even be evaluated in humans, let alone proven effective. Here, we use resveratrol as an example to highlight the enormous difficulties in understanding pharmacokinetics, determining side effects, and, ultimately, establishing mechanisms of action for a natural compound. Despite extensive interest and effort, and continuing promising results from basic science groups, very little is known even today about the effects of resveratrol in humans. Part of PAK6 the problem is the unattractiveness

of natural compounds to large, well-funded companies that could run clinical trials because developing their own molecules affords much greater protection for their intellectual property. In fact, selling unpatentable material motivates smaller nutraceutical companies to complicate the scientific problem even more each creates its own proprietary blend, making it extremely difficult to compare their data with those of other companies, or of academic labs using pure compounds. But even beyond these problems lies a deeper one; resveratrol, and almost every natural compound, is likely to have many clinically relevant targets with different dose response profiles, tissue distributions, and modifiers.

The mechanism is independent of membrane potential We have propo

The mechanism is independent of membrane potential. We have proposed that feedback

inhibition of CcO by ATP keeps LY294002 datasheet the membrane potential and ROS production at low levels. Various forms of stress switch off allosteric ATP-inhibition via reversible dephosphorylation of CcO, resulting in increased membrane potential and cellular ROS levels. This mechanism is proposed to represent a missing molecular link between stress and degenerative diseases.”
“After clinical resolution of signs and symptoms of mild traumatic brain injury (MTBI) it is still not clear if there are residual abnormalities of structural or functional brain networks. We have previously documented disrupted interhemispheric functional connectivity in ‘asymptomatic’ concussed individuals during the sub-acute phase of injury. Testing of 15 normal volunteers (NV) and 15 subacute MTBI subjects was performed within 24 h of clinical symptoms resolution and medical clearance for the first stage of aerobic activity. In this MRS study we report: (a) both in the genu and splenium of the corpus callosum NAA/Cho and NAA/Cr ratios were significantly (p < 0.05) lower in MTBI subjects shortly after the injury compared to NVs, and (b) the metabolic ratio NAA/Cho in the splenium significantly correlated with the magnitude

of inter-hippocampal functional connectivity KPT-330 datasheet in normal volunteers, but not in MTBI. This novel finding supports our

Bacterial neuraminidase hypothesis that the functional disruption of interhemispheric brain networks in MTBI subjects results from compromised metabolic integrity of the corpus callosum and that this persists despite apparent clinical return to baseline. (C) 2011 Elsevier Ireland Ltd. All rights reserved.”
“Aims: To determine whether Clostridium botulinum neurotoxin (BoNT) production in anaerobic culture was affected by temperature and could influence the sandwich ELISA (sELISA) detection of group III toxins in pre-enriched gastrointestinal (GI) contents from clinically suspect cattle botulism cases.

Methods and Results: Bovine post-mortem GI samples taken from 124 and 96 animals with suspect and nonsuspect botulism, respectively, were pre-enriched anaerobically at 30 and 37 degrees C prior to testing by sELISA. After enrichment at 37 degrees C, BoNT was demonstrated in all clinically suspect bovine botulism cases that had been identified by the mouse bioassay, and enrichment by both temperatures enabled BoNT detection in a number of mouse bioassay-negative suspect cases.

Conclusions: Culture temperature does influence the production of group III BoNT, and incubation at both 30 and 37 degrees C is required for optimum detection.

Blocking RAGE by anti-RAGE immunoglobulin G or its silencing by s

Blocking RAGE by anti-RAGE immunoglobulin G or its silencing by siRNA significantly protected podocytes from AOPPs-induced apoptosis both in vitro and in vivo and ameliorated albuminuria in AOPPs-challenged mice. AOPPs-induced activation of nicotinamide adenine dinucleotide phosphate oxidase and the excessive generation of intracellular superoxide were largely inhibited by anti-RAGE immunoglobulin G or LY294002 RAGE siRNA. Moreover, blockade of RAGE decreased the activation of the p53/Bax/caspase-dependent proapoptotic pathway induced by AOPPs. Thus, AOPPs

interact with RAGE to induce podocyte apoptosis and this, in part, may contribute to the progression of chronic kidney disease. Kidney International (2012) CUDC-907 supplier 82, 759-770; doi:10.1038/ki.2012.184; published online 23 May 2012″
“In 2008, a successful computational design procedure was’ reported that yielded active enzyme catalysts for the Kemp elimination.

Here, we studied these proteins together with a set of previously unpublished inactive designs to determine the sources of activity or lack thereof, and to predict which of the designed structures are most likely to be catalytic. Methods that range from quantum mechanics (QM) on truncated model systems to the treatment of the full protein with ONIOM QM/MM and AMBER molecular dynamics (MD) were explored. The most effective procedure involved molecular dynamics, and a general MD protocol was established. Substantial deviations from the ideal catalytic geometries were observed for a number of designs. Penetration of water into the catalytic site and insufficient residue-packing around the active site are the main factors that can cause enzyme designs to be inactive. Where in the past, computational new evaluations of designed enzymes were too time-extensive for practical

considerations, it has now become feasible to rank and refine candidates computationally prior to and in conjunction with experimentation, thus markedly increasing the efficiency of the enzyme design process.”
“Transmission of human cytomegalovirus (HCMV) to the fetus is the most common type of intrauterine infection; the mechanism of HCMV pathogenesis in the developing central nervous system remains unclear. Thrombospondins 1 and 2 (TSP1, TSP2) produced by immature astrocytes are critical for fetal synaptogenesis. To examine the effect of HCMV on fetal astrocytes, human fetal astrocytes were isolated and cultured with HCMV AD169. Cells were harvested at different time points. Protein and mRNA expressions of TSP1 and TSP2 were determined using RT-qPCR, western blotting analysis, and enzyme-linked immunosorbent assay. The results showed that HCMV infection induced time-dependent decreases in mRNA and protein expressions of both TSP1 and TSP2 in astrocytes.

The silent period was longer in the patients than in the controls

The silent period was longer in the patients than in the controls when a RMTh-related SI was used and did not differ between the two groups when a fixed SI was used. We concluded that the observed TMS changes could be interpreted as primary alterations of intracortical motor excitability followed by defects of cortical inhibition and should be attributed to schizophrenia, antipsychotic medication or the interaction between the two factors. (C) 2009 Elsevier Ireland Ltd. All rights reserved.”

prosopagnosia is a condition that, present from an early age, makes it difficult for an individual to recognise someone from APR-246 molecular weight his or her face. Typically, research into prosopagnosia has employed static images that do not contain the extra information we can obtain from moving faces and, as a result, very little is known about the role of facial motion for identity processing in prosopagnosia.

Two experiments comparing the performance of four congenital prosopagnosics with that of age matched and younger controls on their ability to learn and recognise (Experiment 1) and match (Experiment 2) novel faces are reported. It was found that younger controls’ recognition memory performance increased with dynamic presentation, however only one of the four prosopagnosics selleck compound showed any improvement. Motion aided matching performance of age matched controls and all prosopagnosics. In addition, the face inversion effect, an effect that tends to be reduced in prosopagnosia, emerged when prosopagnosics matched moving faces. The results suggest during that facial motion can be used as a cue to identity, but that this may be a complex and difficult cue to retain. As prosopagnosics performance improved with the dynamic presentation of faces it would appear that prosopagnosics can use motion as a cue to recognition, and the different patterns for the face inversion effect that occurred in the prosopagnosics for static and dynamic faces suggests that the

mechanisms used for dynamic facial motion recognition are dissociable from static mechanisms. (C) 2013 Elsevier Ltd. All rights reserved.”
“Endophenotypes have emerged as an important concept in the study of schizophrenia. Perceptual/attentional anomalies were examined as potential endophenotypes in a family study using a strategy for “”multiplex/simplex schizophrenia”". The sample was comprised of 797 subjects: 206 schizophrenia patients, 302 first-degree relatives and 289 controls. The Spanish versions of the Structured Interview for Assessing Perceptual/attentional Anomalies (SIAPA) and Positive and Negative Symptoms Scale (PANSS) were applied to measure the presence of perceptual/attentional anomalies, and positive and negative subscale respectively. An ANCOVA was carried out for global comparisons between groups.

These CTLs (notably CD8(+) T cells) recognize and kill insulin-se

These CTLs (notably CD8(+) T cells) recognize and kill insulin-secreting pancreatic

beta cells, reducing their number PD0325901 purchase by similar to 90%. The resulting reduction of insulin secretion causes the defective regulation of glucose metabolism, leading to the characteristic symptoms of diabetes. Recognition of beta cells as targets by CTLs depends on the interactions between MHC-peptide complexes on the surface of beta cells and receptors (TCRs) on T cells. Those CTLs with high affinity TCRs (also called high avidity T cells) cause most of the harm, while those with low affinity TCRs (also called low avidity T cells) play a more mysterious role. Recent experimental evidence suggests that low avidity T cells accumulate as memory T cells during the disease and may be protective in NOD mice (a strain prone to developing T1D), delaying disease progression. It has been hypothesized that such low avidity T cells afford disease protection either by crowding the islets of Langerhans, where beta cells reside, or by killing antigen presenting cells (APCs).

In this paper, we explore the hypothesized mechanisms for

this protective effect in the context of a series of models for (1) the interactions of low and high avidity T cells, (2) the effect of APCs and (3) the feedback from beta cell killing to autoantigen-induced T cell proliferation. We analyze properties of these models, noting consistency of predictions with observed behaviour. We then use the this website models to examine the influence of various treatment strategies on the progression of the Mannose-binding protein-associated serine protease disease. The model reveals that progressive accumulation of memory low avidity

autoreactive T cells during disease progression makes treatments aimed at expanding these protective T cell types more effective close to, or at the onset of clinical disease. It also provides evidence for the hypothesis that low avidity T cells kill APCs (rather than the alternate hypothesis that they crowd the islets). Crown Copyright (C) 2008 Published by Elsevier Ltd. All rights reserved.”
“Research in songbirds shows that singing behavior is regulated by both brain areas involved in vocal behavior as well as those involved in social behavior. Interestingly, the precise role of these regions in song can vary as a function of the social, environmental and breeding context. To date, little is known about the neurotransmitters underlying such context-dependent regulation of song. Dopamine (DA) modulates highly motivated, goal-directed behaviors (including sexually motivated song) and emerging data implicate DA in the context-dependent regulation of singing behavior. This study was performed to begin to examine whether differences in DA receptors may underlie, in part, context-dependent differences in song production.

In contrast, KSPs bound fewer SDS molecules, and showed a very di

In contrast, KSPs bound fewer SDS molecules, and showed a very different migration time and peak pattern mTOR inhibitor in CE, thereby providing some insight about the structural heterogeneity of SDS:protein complexes and the relative KS of the various proteins.”
“Quercetin has demonstrated protective effects against A beta-induced toxicity on both neurons and endothelial cells. However, whether or not quercetin has an effect on the neurovascular coupling is unclear. In the present study, we aim to investigate the anti-amnesic effects

of quercetin and to explore the underlying mechanisms. A beta(25-35) (10 nmol) was administrated to mice i.c.v. Quercetin was administrated orally for 8 days after injection. Learning and memory behaviors were evaluated by measuring spontaneous alternation in Morris Water Maze test and the step-through positive avoidance test. The regional cerebral find more blood flow was monitored before the A beta(25-35) injection and on seven consecutive days after injection. Mice were sacrificed and cerebral cortices were isolated on the last day. The effects of quercetin on the neurovascular unit (NVU) integrity, microvascular function and cholinergic neuronal changes, and the modification of signaling pathways were tested. Our results demonstrate that quercetin treatment for A beta(25-35)-induced amnesic mice improved Bay 11-7085 the learning and memory

capabilities and conferred robust neurovascular coupling protection, involving maintenance of the NVU integrity, reduction of neurovascular oxidation, modulation of microvascular function, improvement of cholinergic system, and regulation of neurovascular RAGE signaling pathway and ERK/CREB/BDNF pathway. In conclusion, in A beta(25-35)-induced amnesic mice, optimal doses of quercetin administration were beneficial. Quercetin protected the NVU likely through reduction of oxidative damage, inactivation of RAGE-mediated pathway and preservation of cholinergic

neurons, offering an alternative medication for Alzheimer’s disease. (C) 2012 Elsevier Ltd. All rights reserved.”
“Objective: To retrospectively assess the therapeutic value of endovascular stenting for treatment of the nutcracker syndrome (NCS) in long-term follow-up and to explore the selection of the size of stents in Chinese patients with NCS.

Methods: From January 2004 to August 2010, 30 patients (two women and 28 men) between 13 and 32 years old (mean, 18.2) who were diagnosed with NCS were admitted for endovascular treatment. Each patient received one self-expanding metallic stent (14-mm diameter, 60 mm long) in the compressed portion of the left renal vein during the operation, and three patients with severe left-sided varicoceles received left gonadal vein embolization. The postoperative follow-up was 12 to 80 months (median, 36.0 months).

e that Lys-N peptide electron transfer dissociation spectra are

e. that Lys-N peptide electron transfer dissociation spectra are perfectly suited for de novo interpretation. A stringent “”golden”" dataset of peptides identified by conventional database search algorithms was taken to validate the performance of LysNDeNovo. The results on this dataset indicate that LysNDeNovo was able to confidently identify a considerable proportion (42%), without requiring any prior genome or protein sequences. Results of similar quantity and quality could also be obtained

SN-38 price on a much more extensive experimental dataset, illustrating the potential for higher throughput de novo sequencing using these methods.”
“In earlier earlier studies it has been found that rats respond to intracerebroventricular (i.c.v.) injection of cholecystokinin-octapeptide (CCK-8) with a febrile response characterized by rises of heat production and core temperature together with tail-skin Y27632 vasoconstriction mediated by CCK2 receptors. Biotelemetric investigations of the same species have additionally shown that CCK-induced fever is accompanied by decreased locomotor activity.

Similar data for mice have not been reported so far. In the present studies C57BL/6 mice were infused i.c.v. for 3 days with CCK-8 to see effects on body core temperature, locomotor activity, food intake and body weight. Biotelemetric monitoring disclosed a rise in daylight core temperature and a fall of night-time locomotor activity both lasting beyond the time of i.c.v. infusions. Food intake was suppressed only during infusion, while a significant decrease of body weight was sustained after the end of CCK-8 infusion. It is concluded that similar to rats mice also respond to i.c.v. infusion of CCK-8 with a fever-like (regulated) hyperthermia and some components of sickness behavior as measured by biotelemetry, Aspartate and

thus a CCK-mediated mechanism may contribute to fever genesis also in mice. (C) 2011 Elsevier Ltd. All rights reserved.”
“Background: This study aimed to explore the developmental effects of prenatal exposure to Pb and Cd on infant cognitive development at 6 months of age.

Methods: Between 2006 and 2010, the blood levels of Pb and Cd were measured in 884 mothers during their early and late pregnancy. The mental (MDI) and psychomotor (PDI) development index scores of the infants were assessed using the Bayley Scales of Infant Development. The development index scores were adjusted for birth weight, maternal age, maternal education level, family income, breastfeeding status, and residential area.

Results: The geometric mean of the maternal blood concentration was 1.36 mu g/dL (10th percentile = 0.83; 90th percentile = 2.13; range = 0.26-9.10) for Pb and 1.42 mu g/L (10th percentile = 1.01; 90th percentile = 2.16; range = 0.03-9.87) for Cd during the early pregnancy period and 1.27 mu g/dL (10th percentile = 0.77; 90th percentile = 2.10; range = 0.12-4.28) for Pb and 1.52 mu g/L for Cd (10 percentile = 1.07; 90th percentile = 2.10; range = 0.