Quality appraisal was conducted using the Critical Appraisal Skil

Quality appraisal was conducted using the Critical Appraisal Skills Programme. A meta-ethnographic approach was used to categorize further the main findings under key concepts.\n\nResultsSome 1174 citations were retrieved through an electronic database search, reference lists and known literature. Of these, 26 were included for review

after application of full-text inclusion and exclusion criteria. The overall quality of the studies was rated as average to good, with 77 per cent of the methodological quality ACY-738 assessment criteria being met. Six key concepts were identified: signs of effective communication, signs of communication problems, effects on teamwork, conditions for communication, effects on patient safety and understanding collaborative work.\n\nConclusionCommunication was shown to affect operating theatre practices in all of the studies reviewed. Further detailed observational research is needed to gain a better understanding of how to improve the working environment and patient safety in theatre.”
“Discordance in estrogen receptor and human epidermal growth GANT61 price factor receptor 2 receptor status between the primary tumor and recurrence is frequently reported in the literature. This is frequently interpreted as evidence for a change in the biology of breast cancer during the course of the disease. This

commentary discusses some of the caveats of this interpretation. Discordant receptor results can be caused by any of 3 factors: (a) a genuine switch in the biology of the disease, (b) sampling error in focally receptor-positive cancers, and (c) limited accuracy and reproducibility of receptor assays. The relative contribution of each of these factors to discordant results is unknown. A switch in molecular class between primary and recurrent cancer (or residual cancer after therapy) appears to be a rare event based on the available

limited molecular profiling data. Small pockets of strongly focally receptor-positive tumor nests in a larger receptor-negative cancer are also relatively infrequently seen. Discordance resulting from inherent limitations in assay reproducibility is evident from the frequently discordant receptor results even when the same samples are assessed in different laboratories (e. g., central versus local laboratory). A repeat tumor Cl-amidine concentration biopsy is clearly justified when it is suspected, on clinical grounds, that the original receptor results may have been false negative or when the diagnosis of metastatic disease is in question. However, routine repeat biopsy for receptor re-evaluation does not necessarily improve diagnostic accuracy and have a potential to harm through a false-negative result. For patients with clinical courses consistent with hormone responsiveness, or with prior positive hormone receptor results, a course of endocrine therapy is reasonable regardless of the most recent hormone receptor assay result.

Results: Based on classification and regression tree analysis

\n\nResults: Based on classification and regression tree analysis, both the whole limb perometer volume percentage change >= 15% and the sum of circumferences (of 6 defined sites along the limb) percentage change >= 7% performed well overall in predicting moderate or severe perceived swelling (defined as “lymphedema”). Both definitions predicted lymphedema in approximately the same fraction of patients with misclassification rates of 16% and

15%, sensitivity 56% and 50%, specificity 95% and 100%, respectively. Using >= 15% of whole perometer volume BAY 63-2521 percentage change, 12% of patients with inguinal dissection had lymphedema compared with 23% of patients with ilio-inguinal dissection. Combining both groups, 18% of patients had lymphedema, positive and negative predictive values 82% and 84%. Using the definition >= 7% of the sum of circumferences percent change, 7% of patients with inguinal dissection had lymphedema compared with 19% of patients with ilio-inguinal dissection (overall 14% had lymphedema, positive and negative predictive values 100% and 82%, respectively). Of the variables assessed, only radiotherapy was significantly associated with predicted lymphedema (OR 12.6; 95% CI 1.7 to > 100; P = 0.001 using whole perometer

www.selleckchem.com/products/ipi-145-ink1197.html change >= 15%; and OR 13.0; 95%CI 1.4 to > 100; P = 0.021 using sum circumference change >= 7%).\n\nConclusions: A whole limb perometer volume percentage change of >= 15%

and increase in the sum of circumferences of the defined points along the limb >= 7% provide robust definitions of lower limb lymphedema.”
“PACAP is a critical regulator of long-term catecholamine secretion from the adrenal medulla in vivo, however the receptor or pathways for Ca2+ entry triggering acute and sustained secretion have not been adequately characterized. We have previously cloned the selleckchem bovine adrenal chromaffin cell PAC1 receptor that contains the molecular determinants required for PACAP-induced Ca2+ elevation and is responsible for imparting extracellular Ca2+ influx-dependent secretory competence in PC12 cells. Here, we use this cell model to gain mechanistic insights into PAC1hop-dependent Ca2+ pathways responsible for catecholamine secretion. PACAP-modulated extracellular Ca2+ entry in PC12 cells could be partially blocked with nimodipine, an inhibitor of L-type VGCCs and partially blocked by 2-APB, an inhibitor and modulator of various transient receptor potential (TRP) channels. Despite the co-existence of these two modes of Ca2+ entry, sustained catecholamine secretion in PC12 cells was exclusively modulated by 2-APB-sensitive Ca2+ channels. While IP3 generation occurred after PACAP exposure, most PACAP-induced Ca2+ mobilization involved release from ryanodine-gated cytosolic stores.

This study demonstrates that when sampling leaf disks for gene ex

This study demonstrates that when sampling leaf disks for gene expression analysis, the time between tissue removal from the plant and flash freezing in liquid nitrogen can be extended. This increase in time allowable during sampling provides greater flexibility in sampling

large replicated field experiments for statistical analysis of gene expression data.”
“Species of the genus Serrapinnus from the rio Tocantins-Araguaia basin are revised and three new species are described. Serrapinnus aster new species is diagnosed by the presence of check details scimitar-shaped ventral procurrent caudal-fin rays of mature males forming a semicircle and by the presence of 7-9 cusps on the premaxillary teeth; S. lucindai new species is distinguished from its congeners by the presence of a higher number of ventral procurrent caudal-fin rays (17 to 19); and S. tocantinensis new species differs from the remaining species of the genus by the elongation of the unbranched

dorsal and pelvic-fin rays into filaments in mature males. Serrapinnus sterbai is recognized as broadly distributed in the Tocantins-Araguaia basin and is redescribed based on specimens from across its entire distribution. A key for the cheirodontines occurring in the Atlantic drainages of northeastern Brazil, from the rio Tocantins-Araguaia to the rio Paraguacu is provided.”
“The aim of this study was to assess the incidence of stress urinary buy PD0332991 incontinence (SUI) following laparoscopic sacrocolpopexy in continent women and its relation to postoperative prolapse stage. A total of 220 continent women with symptomatic apical prolapse who underwent laparoscopic sacrocolpopexy were prospectively evaluated; 100 women

had previous hysterectomy. Patients were followed up at 3 and 12 months after surgery. All subjects completed the King’s Health Questionnaire AZD0530 supplier (KHQ) and Prolapse Quality of Life Questionnaire (P-QOL) and were examined using the Pelvic Organ Prolapse Quantification (POP-Q) system. The primary outcome measure was the number of women who developed new onset moderate/severe SUI at 3 and 12 months postoperatively and its relation to postoperative prolapse stage. At 3 months, 52 women (23.6 %) developed moderate/severe SUI; 27 (12.2 %) had severe SUI. Eleven women (5.0 %) underwent surgery for SUI within 6 months of sacrocolpopexy. All surgery for SUI was in women who had post-hysterectomy sacrocolpopexy. Postoperatively, the vaginal apex (point C) was at stage 0/I in 195 cases (88.6 %). There were no differences in postoperative POP-Q stage of the anterior and apical vaginal walls between continent women and those with SUI (p = 0.45). The posterior vaginal wall was higher in women who developed de novo SUI (p = 0.03). The incidence of SUI following apical prolapse repair is 23.6 %. Subsequent continence procedures were performed in 5.0 % of patients. All were in women who had previous hysterectomy making the risk in this group 11 %.

Material and method: These guidelines establish some objectiv

\n\nMaterial and method: These guidelines establish some objective and measurable evaluation criteria related to the aspects that intervene in the SLNB. Each criterion is associated

to 4 levels of quality standards. There are 3 essential criteria of mandatory fulfillment to implement SLNB in the clinical practice. They evaluate care experience (6 criteria), operative aspects (18 criteria, 3 essential) and scientific activity (8 indicators). The initial validation phase included 50 patients and the application phase 120 patients (February 2009).\n\nResults: Three essential criteria related to the detection of the sentinel node (98%), accredited sensitivity (90%), and mean number of axillary sentinel nodes (2.1) were fulfilled. The application of

LY3039478 in vivo self-evaluation guidelines in our Unit obtained a global score of 39 out of 71 points, quality level II being the most prevalent\n\nConclusion: During the validation phase, our unit fulfilled the criteria established to implement the procedure. Following that, the use of this guideline accredits the good practice of the SLNB procedure and makes it possible to identify the areas of improvement, in our case, those related to the scientific and training activities. (C) 2010 Elsevier Espana, S.L. and SEMNIM. All rights reserved.”
“Increasing the ability to store mass-reared natural enemies during periods or seasons of low demand is a critical need of the biocontrol industry. We tested the hypothesis that chemicals

can enhance long-term Entinostat ic50 cold storage of a predatory see more mite Phytoseiulus persimilis Athias-Henriot. The research objective was to determine the effect of cryoprotectant and carbohydrate chemicals on in-storage survival of predators. In-storage survival at 8 degrees C was greater for predators sprayed with glycerol (5%, v/v) or glucose (10% and 20%, v/v) than with water spray controls. After 74 days in the cryoprotectant experiment, predator survival declined to 11.5% in the 5% glycerol treatment and 7.8% in the water spray control. After 88 days in the carbohydrate experiment, predator survival declined to 22% in the 20% glucose treatment and 2% in the water spray control. Although many individuals expired within 50 days in both experiments, a few females survived more than 200 days. This research suggests that select cryoprotectants and carbohydrates have a limited capacity to facilitate long-term storage of P. persimilis.”
“Although executive functions (e.g., response inhibition) are often thought to interact consciously with reward, recent studies have demonstrated that they can also be triggered by unconscious stimuli. Further research has suggested a close relationship between consciously and unconsciously triggered response inhibition. To date, however, the effect of reward on unconsciously triggered response inhibition has not been explored.

64 and 0 27, respectively), whereas the domain score urinary inco

64 and 0.27, respectively), whereas the domain score urinary incontinence was inversely correlated (r (s) = -0.32 and -0.35, respectively).\n\nThe sensation or visualization of a bulge in the vagina was the only symptom which correlated positively with the degree of pelvic organ prolapse, and clinical examination and dynamic MR imaging showed similar correlation in this respect.”
“Plant-carbon inputs SBI-0206965 manufacturer to soils in the form of dissolved sugars, organic acids and amino acids fuel much of heterotrophic microbial activity belowground. Initial residence times of these compounds in the soil solution are on the order

of hours, with microbial uptake a primary removal mechanism. Through microbial biosynthesis, the dissolved compounds become dominant precursors for formation of stable soil organic carbon. How the chemical class (e.g. sugar) of a dissolved compound influences stabilization in field soils is unknown and predictions from our understanding of microbial metabolism, AS1842856 turnover and identity are contradictory. We show that soil carbon formation, from chronic amendments of dissolved compounds to fertilized and unfertilized grasslands, is 2.4-times greater from a sugar than an amino acid. Formation rates are negatively correlated with respiration rates of the compounds, and

positively correlated with their recovery in microbial biomass. These relationships suggest that the efficiency of microbial growth on a compound is positively related to formation rates of soil organic carbon. Fertilization does not alter these findings, but together nitrogen and phosphorus additions

reduce soil carbon formation. Our results highlight the need to consider both nutrient enrichment and global-change induced shifts in the form of dissolved root inputs to soils to predict future soil carbon stocks and hence phenomena such as climate warming and food security to which these stock sizes are intimately tied.”
“Measured selleck compound uptake of cyantraniliprole (3-bromo-1-(3-chloro-2-pyridinyl)-N-[4-cyano-2-methyl-6-[(methylamino)-carbonyl]phenyl]-1H-pyrazole-5-carboxamide) into tomatoes following hydroponic exposure allowed calibration of a novel soil uptake model. The total mass of plant parts in treated plants was derived from the weights of successively harvested control plants (no cyantraniliprole provided) over 18 days following the first sampling of ripe tomatoes. Transpired water measured during plant growth was coupled with the calculated increase in plant mass to determine a transpiration coefficient constant (L/kg plant fresh weight) for use in the model. Cyantraniliprole concentrations in mature fruit, fresh foliage, and plant uptake solutions were used as the basis for a nonlinear least squares optimization that consistently resolved to values that were empirically valid compared to metabolism studies in whole plants.


“TIBA is a tool to reconstruct phylogenetic trees from rea


“TIBA is a tool to reconstruct phylogenetic trees from rearrangement data that consist of ordered lists of synteny blocks (or genes), where each synteny block is shared with all of its homologues in the input genomes. The evolution of these synteny blocks, through rearrangement operations, is modelled by the uniform Double-Cut-and-Join model. Using a true distance estimate

under this model and simple distance-based methods, TIBA reconstructs a phylogeny of the input genomes. Unlike any previous tool for inferring phylogenies from rearrangement data, TIBA uses novel methods of robustness estimation to provide support values for the edges in the inferred tree.”
“Background/Aims: To investigate

www.selleckchem.com/products/10058-f4.html the morphological changes of the inner retinal layer by spectral-domain optical coherence tomography (OCT) after idiopathic full-thickness macular hole (MH) surgery. Methods: In a retrospective selleck chemicals llc study, the authors evaluated 52 eyes of 49 patients with MH closed following vitrectomy. All patients were followed postoperatively for more than 6 months. Cross-sectional and retinal surface images were obtained using Cirrus high-definition OCT before and after surgery. In 24 of the 52 eyes, fundus autofluorescence (FAF) was analyzed. Results: The incidence of dissociated optic nerve fiber layer (DONFL) increased gradually over time after surgery. 57.7% had

defects of only the retinal nerve fiber layer (RNFL) and 30.8% had defects in the inner plexifornn layer at 6 months after surgery. Postoperative selleck compound best-corrected visual acuity did not differ significantly based on the depth of the DONFL (p = 0.299). There were no changes in FAF in the area with DONFL. Conclusions: DONFL is characterized by progressive defects that are not limited to RNFL thickness. The healing process after vitrectomy for MH is not limited in the RNFL affecting deeper structural changes. Further investigations are required to evaluate the pathophysiological mechanism of inner retinal change after MH surgery. Copyright (C) 2013 S. Karger AG, Basel”
“Background: Worldwide diversity of alleles of D4 receptor gene (DRD4), linked to attention deficit hyperactivity disorder (ADHD), is mostly the result of length and single nucleotide polymorphisms in a 48-bp tandem repeat (VNTR). Alleles containing from two (2R) to eleven (11R) repeats have been identified. The most common are 4R, 7R and 2R. Aim: To study the association of ADHD risk with DRD4 genotypes in Chilean students.


“Periodontal disease is a localised


“Periodontal disease is a localised KPT-8602 clinical trial inflammatory response caused by the infection of a periodontal pocket arising from the accumulation of subgingival plaque. Periodontal disease has been considered as a possible risk factor for other systemic diseases such as cardiovascular diseases and pre-term low birth weight infants. Advances in understanding the aetiology, epidemiology and microbiology of periodontal pocket flora have revolutionised the therapeutic strategies for the management of periodontal disease progression. This review summarises the recent developments in

the field of intra-pocket drug delivery systems and identifies areas where further research may lead to a clinically effective intra-pocket delivery system.”
“In humans, dehydroepiandrosterone (DHEA), with its sulfate, is the most abundant adrenal steroid, whereas the rat adrenals are not capable of synthesizing this steroid. Circulating concentrations of DHEA sulfate lie in the millimolar range and those of DHEA in the subnanomolar

range. DHEA exerts protective potential during vascular remodeling, Ro-3306 chemical structure although the underlying mechanisms of this protection are imperfectly defined. We hypothesized that physiological doses of DHEA alter signaling pathways that are of central importance for vascular integrity. We exposed human endothelial cells, vascular smooth muscle cells, and fibroblasts to DHEA (10(-6) to 10(-10) mol/L) and observed

a dose-and time-dependent increase of extracellular signal-regulated kinases 1 and 2 activation. Similar results were observed in rat vascular smooth muscle cells. In addition, in rat vascular smooth muscle cells, we found altered phosphorylation and cellular translocation of the transcription factor FoxO1. Pharmacological blockade of the mineralocorticoid receptor Roscovitine (MR) with eplerenone or small interfering RNA-mediated MR-silencing prevented DHEA-induced extracellular signal-regulated kinase 1/2 phosphorylation and its effects on FoxO1. Of note, in a cell-based MR transactivation assay, we did not find any agonist effect of DHEA on MR activity. We conclude that DHEA induces early signaling events in vascular cells that might underlie the DHEA-mediated protection against vasculopathies. These effects are dependent on the MR, although the finding that DHEA fails to act as a direct MR agonist suggests that additional signaling proteins are involved. In this regard, DHEA may either interact with coeffectors to modify MR activity or serves as a ligand for a yet unknown receptor that might transactivate the MR. (Hypertension. 2011;58:471-478.) .

Results Median (interquartile range) of des-acyl ghrelin level,

Results. Median (interquartile range) of des-acyl ghrelin level, was 167 (121-195) pg/ml in HF versus 149 (130-223) pg/ml in post-HTx, p = NS. Acylated ghrelin level was 76 (51-99) pg/ml

versus 13 (0-30) pg/ml, p smaller than 0.001. Acylated/total Ion Channel Ligand Library cell line ratios were 0.33 (0.20-0.47) versus 0.08 (0-0.13), p smaller than HF 0.001. The correlation between acylated and total ghrelin levels was greater in HF than that in HTx. Acyl ghrelin correlated inversely with body mass index in HF, but not in HTx. Conclusion. Acylated ghrelin and the acylated/total ratio were dramatically higher in HF compared with those in HTx. Acylation rather than secretion of ghrelin is upregulated in HF and the resistance to ghrelin’s anabolic and appetite-stimulating effects is not at the level of acylation, but downstream at the ghrelin-receptor

level.”
“Long interspersed element-1s (LINE-1 or L1s) are abundant retrotransposons check details that occur in mammalian genomes and that can cause insertional mutagenesis and genomic instability. L1 activity is generally repressed in most cells and tissues but has been found in some embryonic cells and, in particular, in neural progenitors. Moreover, L1 retrotransposition can be induced by several DNA-damaging agents. We have carried out experiments to verify whether L1 retrotransposition is affected by oxidative DNA damage, which plays a role in a range of human diseases, including cancer and inflammatory and neurodegenerative disease. To this purpose, BE(2)C neuroblastoma cells, which

are thought to represent embryonic precursors of sympathetic neurons, have been treated with hydrogen peroxide and subjected to an in vitro retrotransposition assay involving an episomal L1(RP) element tagged with enhanced green fluorescent protein. Our results indicate that hydrogen peroxide treatment induces an increase HSP990 in the retrotransposition of transiently transfected L1(RP) and an increase in the expression of endogenous L1 transcripts. An increase of gamma-H2AX foci and changes in the mRNA levels of MRE11, RAD50, NBN and ERCC1 (all involved in DNA repair) have also been found. Thus, oxidative stress can cause L1 dysregulation.”
“Decellularized bone/bone marrow was prepared to provide a microenvironment mimicking that of the bone marrow for three-dimensional culture in vitro. Bone/bone marrows were hydrostatically pressed at 980 MPa at 30 degrees C for 10 min to dismantle the cells. Then, they were washed with EGM-2 and further treated in an 80% EtOH to remove the cell debris and lipid, respectively. After being rinsed and shaken with PBS again, treated bone/bone marrows were stained with hematoxylin and eosin (H-E) to assess the efficacy of decellularization.

To date, prognosis

for patients with these tumors is base

To date, prognosis

for patients with these tumors is based on results from small single-center patient series and controlled trials. Large population-based observational studies are lacking. To assess national trends in histology-specific survival, the authors reviewed patient survival data spanning 30 years (1973-2003) CH5183284 nmr from the Surveillance, Epidemiology, and End Results (SEER) registry, a US population-based cancer registry.\n\nMethods. The SEER registry was queried to identify cases of histologically confirmed primary spinal chordoma, chondrosarcoma, osteosarcoma, or Ewing sarcoma using coding from the International Classification of Disease for Oncology, Third Edition. Association

of survival with histology, metastasis status, tumor site, and year of diagnosis was assessed using Cox proportional-hazards regression analysis.\n\nResults. A total of 1892 patients were identified with primary osseous spinal neoplasms (414 with chordomas, 579 with chondrosarcomas, 430 with osteosarcomas, CX-6258 ic50 and 469 with Ewing sarcomas). Chordomas presented in older patients (60 +/- 17 years; p < 0.01) whereas Ewing sarcoma presented in younger patients (19 +/- 11 years; p < 0.01) compared with patients with all other tumors. The relative incidence of each tumor type remained similar per decade from 1973 to 2003. African Americans comprised a significantly greater proportion of patients with osteosarcomas than other tumors (9.6% vs 3.5%, respectively; p < 0.01). Compared with the sacrum, the mobile spine was more likely to be the site of tumor location for chordomas than for all other tumors (47% vs 23%, respectively; p < 0.05). Osteosarcoma and Ewing sarcoma were 3 times more likely than chondrosarcoma and chordoma to present with metastasis (31% vs 8%, respectively). Resection was performed more frequently for chordoma (88%) and chondrosarcoma (89%) than for osteosarcoma (61%) and PXD101 price Ewing sarcoma (53%). Overall median survival was histology-specific (osteosarcoma, 11 months; Ewing

sarcoma, 26 months; chondrosarcoma, 37 months; chordoma, 50 months) and significantly worse in patients with metastasis at presentation for all tumor types. Survival did not significantly differ as a function of site (mobile spine vs sacrum/pelvis) for any tumor type, but more recent year of diagnosis was associated with improved survival for isolated spinal Ewing sarcoma (hazard ration [HR] 0.95; p = 0.001), chondrosarcoma (HR 0.98; p = 0.009), and chordoma (HR 0.98; p = 0.10), but not osteosarcoma.\n\nConclusions. In this analysis of a 30-year, US population-based cancer registry (SEER), the authors provide nationally representative prognosis and survival data for patients with malignant primary spinal osseous neoplasms.

PAAG deposited extensively in the breast tissues, armpits and spa

PAAG deposited extensively in the breast tissues, armpits and space of the thoracic-abdominal wall, and the breast was connected with the abdominal wall through the fistula of different sizes. At 2 weeks, the percentages of decrease

in Selleck PD-1/PD-L1 Inhibitor 3 drainage volume and in lesion lacuna size of the thoracic-abdominal wall (82% and 80%, respectively) in patients receiving the multiple incisions combined with radical therapy were significantly different from those who did not receive the multiple incisions (46% and 45%) (Both P smaller than 0.01). At 4 weeks, in some of the patients receiving the multiple incisions combined with radical therapy, the lacuna of the thoracic-abdominal wall disappeared Oligomycin A molecular weight completely, and the lesions with flowing masses had been cleared. Conclusions: The new method of subareolar incision combined with surgery for inferior segment of mass to clean the mixture and thoroughly eliminate the lacuna of the thoracic-abdominal wall as well as suture to close the intramammary fistula can improve the treatment efficacy.”
“In solid organ transplantation, human cytomegalovirus (HCMV) is considered to be the most important viral pathogen. We report a case of a CMV R-/D+ small intestine transplant recipient with a primary CMV infection on valganciclovir prophylaxis. Sequencing of the HCMV DNA for drug resistance-associated mutations revealed the UL97 mutation N510S.

This mutation has been initially reported to confer ganciclovir resistance. Based on in vitro recombinant phenotyping, this assumption has recently been questioned. Switching the antiviral treatment to an intravenous regimen MCC950 chemical structure of ganciclovir eliminated HCMV DNAemia, showing the in vivo efficacy of ganciclovir for the UL97 mutation N510S. Hence, knowledge of drug efficacy is crucial for an adequate choice of antiviral medication, carefully balancing antiviral potency versus the risk of harmful side effects.”
“Plasminogen activator inhibitor (PAI)-1 is a major fibrinolytic inhibitor. High PAI-1 is associated with increased

renal and cardiovascular disease risk. Previous studies demonstrated PAI-1 down-regulation by 1,25-dihydroxyvitamin D-3 (1,25(OH)(2)D-3), but the molecular mechanism remains unknown. Here we show that exposure of mouse embryonic fibroblasts to TNF alpha or LPS led to a marked induction of PAI-1, which was blunted by 1,25(OH)(2)D-3, NF-kappa B inhibitor or p65 siRNA, suggesting the involvement of NF-kappa B in 1,25(OH)(2)D-3-induced repression. In mouse Pai-1 promoter a putative cis-kappa B element was identified at -299. EMSA and ChIP assays showed that TNE-alpha increased p50/p65 binding to this kappa B site, which was disrupted by 1,25(OH)(2)D-3. Luciferase reporter assays showed that PAI-1 promoter activity was induced by TNF alpha or LPS, and the induction was blocked by 1,25(OH)(2)D-3.