At baseline, the two groups did not differ significantly with respect to gender, age, BMI and outcome measures. Comparing the differences between groups at short-and mid-term follow-up, the GPR group revealed a significant reduction

(from baseline) Belinostat inhibitor in all outcome measures with respect to the SE group. The ordered logistic regression model showed an increased likelihood of definitive improvement (reduction from baseline of at least 30% in RMDQ and VAS scores) for the GPR group compared to the SE group (OR 3.9, 95% CI 2.7 to 5.7).\n\nConclusions: Our findings suggest that a GPR intervention in subjects with persistent LBP induces a greater improvement on pain and disability as compared to a SE program. These results must be confirmed by further studies with higher methodological standards, including randomization, larger sample size, longer follow-up and subgrouping of the LBP subjects.”
“It has been recognized that new antimalarials with a novel mode of action are critical to combat the continued emergence and dissemination of drug-resistant parasites that threaten the efficacy of current malaria treatments. Thus, recent high-throughput check details screening campaigns have been initiated using asexual intraerythrocytic stage cell-based assays of Plasmodium

falciparum. These have led to the unprecedented identification of over 10,000 new antimalarial compounds. Inherently, novel compounds identified by cell-based assays will have poorly defined modes of action. While some of these compounds may have recognizable targets, the majority of cell-based hits are comprised of unique chemical scaffolds usually lacking cross-resistance with known drugs. It is likely that these novel antimalarial scaffolds will reveal new targets. A challenge for the community will be to assign these small molecules to their targets. In this article, we review methodologies LY2606368 to assist in the determination of a compound’s mode of action.”
“High surface ozone concentration is increasingly being recognized as a factor

that negatively affects crop yields in Asia. However, little progress has been made in developing ozone-tolerant genotypes of rice-Asia’s major staple crop. This study aimed to identify possible tolerance mechanisms by characterizing two quantitative trait loci (QTLs) that were previously shown to influence visible leaf symptoms under ozone exposure (120 nl l(-1), 7 h d(-1), 13 d). Two chromosome segment substitution lines (SL15 and SL41) that carried introgressions of the QTLs OzT3 and OzT9, respectively, were exposed to ozone at 120 nl l(-1) along with their parent Nipponbare. In accordance with the expected QTL effect, SL15 showed stronger visible symptoms of ozone damage than Nipponbare, whereas SL41 had fewer symptoms.

Concerted action between the US and Europe could

selleck move vaccine safety monitoring to today’s level of requirements globally and should be pursued. Published by Elsevier Ltd on behalf of The International Alliance for Biological Standardization.”
“Angiopoietin-1 is a vascular strengthening factor during vascular development and a protective factor for pathological vascular inflammation and leakage. Brain vascular leaking and inflammation are two important pathological processes of stroke; therefore, we hypothesized that variants of the microRNA-binding site in angiopoietin-1 would affect its expression and confer a risk of stroke. To test our hypothesis, a predicted microRNA-binding site was found in the 3′-UTR of angiopoietin-1 using bioinformatics; variant rs2507800 was identified to be located in the miR-211-binding site of angiopoietin-1. Secondly, the effects of the identified variant on angiopoietin-1 translation were assessed using a luciferase reporter assay and ELISA. We found that the A allele of rs2507800 suppressed angiopoietin-1 translation by facilitating miR-211 binding, but not the T allele. Subjects carrying the TT genotype had higher plasma

angiopoietin-1 levels than those with the A allele. Finally, the association of the variant with stroke Lapatinib was tested in 438 stroke patients and 890 controls, and replicated in an independent population of 1791 stroke patients and 1843 controls. The TT genotype resulted in a significant reduction in overall stroke risk OR, 0.51 [95% confidence interval (CI), 0.36-0.74], P = 0.0003, ischemic stroke [OR, 0.56 (95% CI, 0.36-0.85), P = 0.007] and hemorrhagic stroke [OR, 0.46 (95% CI, 0.26-0.80), P = 0.007]. These results were confirmed in an independent study. Our results provide evidence that

the TT genotype (rs2507800) in the 3′-UTR of angiopoietin-1 might reduce the risk of stroke by interfering with miR-211 binding.”
“Enterovirus type 71 (EV71) 2A protease exhibited strong transcriptional activity in yeast cells. The transcriptional activity of 2A protease was independent of Selleck ACY-738 its protease activity. EV71 2A protease retained its transcriptional activity after truncation of 40 amino acids at the N-terminus but lost this activity after truncation of 60 amino acids at the N-terminus or deletion of 20 amino acids at the C-terminus. Thus, the acidic domain at the C-terminus of this protein is essential for its transcriptional activity. Indeed, deletion of amino acids from 146 to 149 (EAME) in this acidic domain lost the transcriptional activity of EV71 2A protein though still retained its protease activity. EV71 2A protease was detected both in the cytoplasm and nucleus using confocal microscopy analysis. Coxsackie virus B3 2A protease also exhibited transcriptional activity in yeast cells. As expected, an acidic domain in the C-terminus of Coxsackie virus B3 2A protease was also identified.

The CdS/CdSe/CdS nanoplatelets represent a new example of colloidal nanoheterostructures with mixed confinement regimes for electrons and holes. In these materials holes are confined to a thin (similar

to 1.8 nm) two dimensional CdSe quantum well, while the electron confinement can be gradually relaxed in all three dimensions by growing epitaxial CdS layers on both Sides of the quantum well find protocol The relaxation of the election confinement energy caused a shift of the emission band from 510 to 665 nm with unusually small inhomogeneous broadening of the emission spectra.”
“As researchers continue to understand non-clinical psychosis (NCP-brief psychotic-like experiences occurring in 5-7% of the general population; van Os et al., 2009), it is becoming evident that functioning deficits and facial emotion recognition (FER) impairment characterize this phenomenon. However, the extent to which these domains are related remains unclear. Social/role functioning and FER were assessed in 65 adolescents/young adults exhibiting low and high-NCP. Results indicate that

FER and social/role functioning deficits were present in the High-NCP group, and that the domains were associated in this group alone. Taken together, findings suggest that a core emotive deficit is tied to broader social/role dysfunction in NCP. (C) 2012 Elsevier B.V. All rights reserved.”
“Ellagic acid has been XMU-MP-1 chemical structure shown to inhibit tumor cell growth. However, the underlying molecular mechanisms remain elusive. In this study, our aim was to investigate whether ellagic acid inhibits the proliferation of MCF-7 human breast cancer cells via regulation of the TGF-beta/Smad3 signaling pathway. MCF-7 breast cancer cells were transfected with pEGFP-C3 or pEGFP-C3/Smad3 plasmids, and treated with ellagic acid alone or in combination with SIS3, a specific inhibitor of Smad3 phosphorylation.

Cell proliferation was assessed by MTT assay and the cell cycle was detected by flow cytometry. Moreover, gene expression was detected by RT-PCR, real-time PCR and Western blot analysis. The selleck chemicals MTT assay showed that SIS3 attenuated the inhibitory activity of ellagic acid on the proliferation of MCF-7 cells. Flow cytometry revealed that ellagic acid induced G0/G1 cell cycle arrest which was mitigated by SIS3. Moreover, SIS3 reversed the effects of ellagic acid on the expression of downstream targets of the TGF-beta/Smad3 pathway. In conclusion, ellagic acid leads to decreased phosphorylation of RB proteins mainly through modulation of the TGF-beta/Smad3 pathway, and thereby inhibits the proliferation of MCF-7 breast cancer cells.”
“Objectives: The aim of this study was to elucidate the prognostic significance of mitral regurgitation (MR) after primary percutaneous coronary intervention (PCI) for acute ST-elevation myocardial infarction (STEMI).

001). However, TH increased phase singularity number (wavebreaks) during VF (P<0.05) and Si pacing (P<0.05). TH resulted in earlier onset of APD alternans (P<0.001), which was predominantly SDA (P<0.05), and increased pacing-induced VF episodes (P<0.05). TH also decreased CV, shortened wavelength, and enhanced APD dispersion and the spatial heterogeneity of CV restitution.\n\nConclusions: TH (30 degrees C) increased the vulnerability of pacing-induced VF by (1) facilitating wavebreaks during VF and Si pacing, and (2) enhancing proarrhythmic electrophysiological parameters, including promoting

earlier onset of APD alternans (predominantly SDA) during selleck chemicals llc S1 pacing. (Circ J 2009; 73: 2214-2222)”
“Brain metastasis has become an increasing cause of

morbidity QNZ supplier and mortality in cancer patients as the treatment of systemic disease has improved. Brain metastases frequently are highly vascularized, a process driven primarily by VEGF. VEGF mediates numerous changes within the vasculature including endothelial cell retraction and increased permeability, vasodilation, and new vessel formation. Here we describe a xenograft brain metastasis model that mimics the critical steps of metastasis including tumor cell dissemination and vascular adhesion, tumor growth and tumor associated angiogenesis. Magnetic resonance (MR) imaging was used to evaluate two aspects of the functional response of brain metastasis to the anti-VEGF receptor therapeutic, AZD2171 (Cediranib, RECENTIN (TM)). MR tracking of individual cells demonstrated that cediranib did not impede tumor

cell extravasation into the brain parenchyma despite evidence that anti-VEGF treatment decreases the permeability of the blood brain barrier. In a second assay, blood volume imaging using ultrasmall superparamagnetic iron oxide revealed that treatment of well-developed brain metastasis with cediranib for 7 days led to a heterogeneous response with respect to individual tumors. Overall, there was a significant average decrease in the tumor vascular bed volume. The majority of large tumors demonstrated substantially reduced central blood volumes relative to normal brain while retaining a rim of elevated blood volume at buy Semaxanib the tumor brain interface. Small tumors or occasional large tumors displayed a static response. Models and assays such as those described here will be important for designing mechanism-based approaches to the use of anti-angiogenesis therapies for the treatment of brain metastasis.”
“Objective: We describe the short-term results of the patients who underwent transapical treatment of a paravalvular leak (PVL) in our centre. Background: Increasing experience with transapical aortic valve implantation has inspired us to explore this approach for prosthetic paravalvular leak reduction in high risk patients.

A high number and variety of microbes, including a high proportion of fungal pathogens, were detected.”
“Purpose: Paclitaxel-coated balloon catheters inhibit restenosis after coronary and peripheral angioplasty (PCI,PTA). The aim of this study was to investigate paclitaxel plasma levels and laboratory parameters following PTA with paclitaxel-coated balloons (PCB) in peripheral arteries.\n\nMaterials and Methods: This single treatment arm, multicenter study included

14 patients with Rutherford stage 1 – 5 with occlusions of up to 5 cm or >= 70% diameter stenosis of the superficial femoral or popliteal arteries (SFA, PA). PTA was performed using up to three PCB catheters. The paclitaxel plasma levels and safety laboratory EPZ-6438 in vitro parameters were determined by collecting blood samples pre-intervention,

immediately post-intervention, at 0.5, 1, 2, 4, 8, 24 hours and 1 and 4 weeks post-intervention (p.i.). Vital signs were monitored to assess clinical safety.\n\nResults: PTA was performed successfully in all patients. Paclitaxel plasma levels were always below a level and duration known to cause systemic side effects. A mean peak paclitaxel plasma level (40 ng/ml) was reached immediately p.i. and decreased rapidly below detectable levels in more than half of the patients already 2 hours p.i. The paclitaxel plasma concentrations returned to values below detectable levels at 24 hours p.i. EVP4593 NF-��B inhibitor in all patients. Laboratory parameters and vital signs did not give any reason for safety concerns. No adverse events associated with balloon PXD101 mw coating were observed.\n\nConclusion:

The results of 14 patients with peripheral arterial occlusive disease show no systemic bioavailability of paclitaxel >24 hours after PTA with one or more PCB catheters, indicating that the PCB catheter is safe with regard to possible systemic effects.”
“Background/Aim: Unresectable metastatic colorectal cancer with very slow tumour growth rate does not necessarily require for strong short-interval chemotherapy. In the present study, we administered monthly chemotherapy and aimed to evaluate the usefulness of the specific treatment schedule in patients with unresectable metastatic colorectal cancer with slow tumour growth rate. Patients and Methods: Since 2009, at our Institution, patients’ whose serum carcinoembryogenic antigen (CEA) values on the treatment day were not higher than those before initial chemotherapy, and patients who did not wish to undergo intensive chemotherapy, were prospectively scheduled for specific chemotherapy. Between January 2009 and December 2011, 10 patients with unresectable metastatic colorectal cancer who received monthly chemotherapy were enrolled in the current study.

(C) 2013 Elsevier Ltd. All rights reserved.”
“Nysius caledoniae Distant, 1920 is recorded for the first

time as an alien introduced species in New Zealand. Collection with young stages indicates that it may already have established itself in Auckland. Characters of the species enabling its identification Hippo pathway inhibitor are presented. The pest-status of N. caledoniae, N. vinitor Bergroth, 1891, and N. huttoni White, 1878, is reviewed. A key to the genera of Orsillidae in New Zealand is provided, along with a revised key to the species of Nysius in that country.”
“This discussion examines how speaker pauses, both filled and silent, are keyed to functions within a conversation and to functions

within narration. In Alzheimer’s discourse, pause-fillers can be both placeholders and hesitation markers; they may be ohs and ums or longer formulaic phrases. Extracts from the speech of 4 older women from the United States and from New Zealand are reviewed for changes in syntactic complexity, for retention of story components, and for pauses. The extracts illustrate these functions for silent pauses: as word-finding; as planning at word, phrase, and narrative component Selleckchem Pevonedistat levels; and as pragmatic compensation as other interactional and narrative skills decrease.”
“Intracellular inclusions containing chlamydia-like organisms are frequently observed in the gill epithelial cells of Atlantic salmon,

Salmo salar L., cultured in fresh water in Ireland. In this study, the causative agent was identified in four separate freshwater sites, using 16s rRNA sequencing, as ‘Candidatus Clavochlamydia salmonicola’. Histopathology and real-time (RT) PCR were used to further assess infections. The prevalence of infection ranged from 75-100% between sites and infection intensity was highly variable. No significant lesions were associated with these infections. As a diagnostic tool, RT-PCR proved marginally more sensitive than this website histopathology. The fate of ‘Candidatus Clavochlamydia salmonicola’ in Atlantic salmon post-seawater transfer was investigated in a 12-week marine longitudinal study. Both RT-PCR and histopathological examination indicate that the organism disappears from the gills 4-6 weeks post-transfer.”
“Stimuli responsive pH- and temperature-sensitive hydrogel drug delivery systems, as those based on N-isopropylacrylamide (NiPAAm) and itaconic acid (IA), have been attracting much of the attention of the scientific community nowadays, especially in the field of drug release. By adjusting comonomer composition, the matrix is enabled to protect the incorporated protein in the highly acidic environment of upper gastrointestinal tract and deliver it in the neutral or slightly basic region of the lower intestine.

In particular, the carboxylic acid groups on a polymer containing L-aspartic acid units were converted to N-hydroxysuccinimidyl esters, providing a useful template for further derivatization. (C) 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47:

3757-3772, 2009″
“Background This study cross-validated the factor structure of the Self-Stigma Scale-Short (SSS-S) in a cohort of patients with mental illness in southern LY294002 clinical trial Taiwan. The measurement invariance of the SSS-S factor structure across mental illness and gender was also examined. Methods The sample consisted of 161 patients with schizophrenia (51.6% males; mean age +/- SD = 40.53 +/- 10.38 years) and 189 patients with other mental illnesses (34.9% males; mean age = 46.52 +/- 11.29 years). Results The internal reliability (total score: alpha = 0.948) and concurrent validity (r = 0.335 to 0.457 with Depression and Somatic Symptoms Scale; r = -0.447 to -0.556 with WHOQOL-BREF)

of the SSS-S were both satisfactory, and the results Selleckchem Nepicastat verified that the factor structure in our Taiwan sample (RMSEA = 0.0796, CFA = 0.992) was the same as that of the Hong Kong population. In addition, the results supported the measurement invariance of the SSS-S across mental illness (Delta RMSEAs = -0.0082 to -0.0037,Delta CFAs = 0.000) and gender (Delta RMSEAs = -0.0054 to -0.0008,Delta CFAs = -0.001 to 0.000). Conclusion Future studies can use the SSS-S to compare self-stigma between genders and between patients with different kinds of mental Dactolisib PI3K/Akt/mTOR inhibitor illnesses.”
“Radiosurgery for glioblastoma is limited to the development of resistance, allowing tumor cells to survive and initiate tumor recurrence. Based on our previous work that coadministration of tissue factor and lipopolysaccharide following radiosurgery selectively induced thrombosis in cerebral arteriovenous malformations, achieving thrombosis of 69% of the capillaries

and 39% of medium sized vessels, we hypothesized that a rapid and selective shutdown of the capillaries in glioblastoma vasculature would decrease the delivery of oxygen and nutrients, reducing tumor growth, preventing intracranial hypertension, and improving life expectancy. Glioblastoma was formed by implantation of GL261 cells into C57Bl/6 mouse brain. Mice were intravenously injected tissue factor, lipopolysaccharide, a combination of both, or placebo 24 hours after radiosurgery. Control mice received both agents after sham irradiation. Coadministration of tissue factor and lipopolysaccharide led to the formation of thrombi in up to 87 +/- 8% of the capillaries and 46 +/- 4% of medium sized vessels within glioblastoma. The survival rate of mice in this group was 80% versus no survivor in placebo controls 30 days after irradiation. Animal body weight increased with time in this group (r = 0.88, P = 0.0001).

“
“We present design of novel low-power homonuclear dipolar recoupling experiments for magic-angle-spinning solid-state NMR studies of proteins. The pulse sequences are developed by combining principles of symmetry-based dipolar recoupling and optimal

control-based pulse sequence design. The scaffold of the pulse sequences is formed by known CN-type recoupling sequences, while the intrinsic sequence elements are designed using optimal control. This procedure allows for the development of high-performance pulse sequences demanding significantly weaker rf fields than previous symmetry-based pulse sequences while compensating for rf inhomogeneity and providing excitation over relevant ranges of chemical shifts for biological applications. The new recoupling experiments,

check details referred to as optimal control CN ((OC)CN), are demonstrated numerically and experimentally by two-dimensional (2D) (13)C-(13)C and three-dimensional (3D) (15)N- (13)C- (13)C chemical shift correlation experiments on uniformly (13)C, (15)N-labeled ubiquitin. Exploiting the double-quantum, band-selective dipolar recoupling properties of the (OC)CN experiments, we demonstrate significant sensitivity enhancement for 2D and 3D correlation spectra showing exclusively one- or two-bond correlations. (C) 2009 American Institute of Physics. [DOI: 10.1063/1.3157737]“
“Global warming is already causing salinization of freshwater ecosystems located in semi-arid regions, including YH25448 clinical trial Turkey. Daphnids, which are important grazers on phytoplankton and a major food source for fish and invertebrates, are sensitive to not only changes in salinity levels, but also presence of predators. In this study, the interactive effect of salinity toxicity (abiotic factor) with predation pressure mimicked by the fish-exuded kairomone (biotic factor) and the effect of salt acclimation on daphnids were investigated. Impacts of these stressors on daphnid survival, life history and

molecular profile were observed. The presence FK228 mouse of the kairomone antagonistically alters the effect of salinity, as observed from the 24- and 48-h LC50 values and survival results. Molecular findings provided solid evidence to this antagonism at even lower salt concentrations, for which antagonism was not evident with organismal data. Fish predation counterbalances the negative effect of salinity in terms of reserve energy density. Therefore, it is important to investigate multiple stressor effects in ecotoxicological bioassays complemented with molecular techniques. The single effect of increasing salinity resulted in increased mortality, decreased fecundity, and slower somatic growth in Daphnia, despite their acclimation to salinity.

Of these, 81 were receiving statins and/or fibrates as LLT and 31 were participating in a non-pharmacologic therapeutic program FK228 manufacturer (NPT) of diet and exercise therapy. The mean age for the total population was 56.7 years (20-78): the LLT group 58.6 and NPT group 51.9 years. Women comprised 53% of the LLT group and 58% of the NPT. No significant differences in baseline lipid profiles, CK level, BMI, waist measurement, gender, cigarette smoking, alcohol consumption, non-steroidal anti-inflammatory drugs or acetaminophen use, frequency of myalgias, SF-36 PF or mHAQ scores were observed between groups. On comparison of gender groups, we ob-served that men receiving LLT had significantly better

SF36 PF (p = 0.037) than men on NPT. There were no differences in SF-36 PF or mHAQ scores between selleck chemicals llc groups for females.\n\nConclusion: We found no adverse effects of LLT on physical functioning or quality of life. Indeed, men treated with LLT had significantly better SF-36 PF scores than men treated non-pharmacologically.”
“The aim of this study was to investigate the oxidative stress status and antioxidant enzyme activities in infertile men’s semen and to determine their relationship with spermatozoa characteristics. Four groups of infertile men, normozoospermic (n = 9), azoospermic (n = 13), oligoasthenozoospermic (n = 38), and

asthenozoospermic (n = 42), were tested for malonaldialdehyde (MDA) concentration and catalase (CAT) and superoxide dismutase (SOD) activities in semen using spectrophotometric methods. We found that MDA levels in semen and SOD activity in seminal plasma (SP) were significantly higher in oligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic group. In contrast, the mean values of CAT activity were not

significantly different in all groups. We found positive correlations between semen MDA concentration and SOD activity (P < 0.01). MDA levels in sperm pellet and in SP were also positively correlated with mobility grade b (P < 0.01), acrosome anomalies (P < 0.01), and residual cytoplasmic droplets (P < 0.05). In contrast, SOD activity in SP was negatively correlated with sperm concentration and normal morphology (P < 0.05). Similarly, CAT activity in SP was negatively correlated with sperm and leukocyte concentrations (P < GDC-0068 0.05). In conclusion, our results suggest that the evaluation of oxidative status and antioxidant defenses may be taken as an important tool for diagnosis and treatment of male infertility. J. Clin. Lab. Anal. 23:99-104, 2009. (C) 2009 Wiley-Liss, Inc.”
“T cell receptor excision circles (TRECs) have been suggested to be useful markers of recent thymic output and graft-versus-host disease (GVHD) was thought to have an adverse effect on thymic output. On the other hand, TREC concentrations are diluted with T cell expansion. The C-type lectin superfamily inhibitory receptor CD94 expression on T cells might be a consequence of CD8 T cell expansion.

These profiles are sampled from dune crests, away from high concentrations of animals and without

termite mounds. Given low-density animal grazing is unlikely to contribute consistent spot-scale nitrate over decades, these profiles give an initial estimate of naturally-produced concentrations. This insight is important for the management of the Stampriet Basin and wider Kalahari groundwater. This study expands our knowledge about elevated nitrate in dryland USZs, demonstrating that it can occur as pulses, probably in response to transient vegetation cover and that it is not limited to long-residence time USZs with very limited downward moisture flux (recharge). (C) 2014 Elsevier Ltd. All rights reserved.”
“The aim of this work is to compare INCB024360 the thinning patterns of the ganglion cell inner-plexiform layer (GCIPL) and peripapillary retinal nerve fiber layer (pRNFL) as measured using Cirrus high-definition optical coherence tomography (HD-OCT) in patients with visual field (VF) defects that respect AZD9291 datasheet the vertical meridian. Twenty eyes of 11 patients with VF defects that respect the vertical meridian were enrolled retrospectively. The thicknesses

of the macular GCIPL and pRNFL were measured using Cirrus HD-OCT. The 5 and 1 % thinning area index (TAI) was calculated as the proportion of abnormally thin sectors at the 5 and 1 % probability level within the area corresponding to the affected VF. The 5 and 1 % TAI were compared between the GCIPL and pRNFL measurements. The color-coded GCIPL deviation map showed a characteristic vertical thinning pattern of the GCIPL, which is also seen in the VF of patients with brain lesions. The 5 and 1 % TAI were significantly higher in the GCIPL measurements than in the pRNFL measurements (all p smaller than 0.01). Macular GCIPL analysis clearly visualized a characteristic topographic pattern of retinal ganglion cell (RGC)

loss in patients with VF defects that respect the vertical BKM120 cost meridian, unlike pRNFL measurements. Macular GCIPL measurements provide more valuable information than pRNFL measurements for detecting the loss of RGCs in patients with retrograde degeneration of the optic nerve fibers.”
“To evaluate clinical and imaging features of cancer recurrence in reconstructed breasts following skin-sparing mastectomy (SSM) or nipple areolar skin-sparing mastectomy (NASSM). This study was approved by our Institutional Review Board. In this retrospective study, we included patients with pathologically confirmed recurrent cancer who had transverse rectus abdominis myocutaneous (TRAM) flap reconstruction after SSM or NASSM and whose follow-up radiological studies were available. Each patient’s demographic data, imaging studies and clinical outcomes were reviewed.