As a result, the M1-M2 polarization balance was increased in FH a

As a result, the M1-M2 polarization balance was increased in FH and, more markedly in NFH. M1 MoMas and the M1-M2 polarization ratio were directly correlated to pre-treatment LDL cholesterol levels and strongly associated with the presence of atherosclerotic plaques. In conclusion, we show for the first time that human hypercholesterolemia is associated with a pro-inflammatory imbalance of circulating monocytic cells, which can predispose to the development of atherosclerosis. (C) 2014 Elsevier Ireland Ltd. All rights reserved.”
“Key message Comparative proteome analysis revealed the proteins that are differentially

expressed during the processes of tetraploid black locust (Robinia pseudoacacia L.) Copanlisib cost adventitious root formation, which are involved in carbohydrate binding, energy and metabolism, and stress response. Abstract The tetraploid black locust (Robinia pseudoacacia L.) is an important fast-growing tree used in afforestation and is also a protein-rich feed for poultry. Currently, the primary approach for tetraploid black locust reproduction is through cutting propagation. To identify the specific proteins in the rooting process, including the formation of adventitious root primordium (ARP) and

Combretastatin A4 inhibitor the elongation of adventitious root (EAR), a comparative proteomic analysis of the ARP and EAR of tetraploid black locust was performed to identify ACY-1215 in vivo related proteins that may be involved in regulating

ARP formation and the process of EAR. High-resolution two-dimensional electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric (MS) analysis was used to identify differentially expressed proteins in ARP and EAR. A total of 84 protein spots showed significant expression changes by 2-DE and were successfully identified by MALDI TOF/TOF MS/MS. Of these, 29 proteins were differentially expressed during the ARP phase, and the remaining 55 proteins did so in the EAR phase. The majority of identified proteins were classified into functional categories, including carbohydrate binding, energy and metabolism, protein degradation/folding and import, cellular cytoskeleton, hormonerelated and oxidation reactions, stress response, cell proliferation, and transcription regulation. Quantitative real-time PCR analysis was performed for selected proteins and showed that not all of the protein expression levels were consistent with the mRNA levels. This study provides the basis for further functional studies of differentially expressed proteins, which will contribute to the understanding of the biochemical processes in adventitious root primordium formation and the elongation of adventitious root.

The microtubule depolymerase mitotic centromere-associated kinesi

The microtubule depolymerase mitotic centromere-associated kinesin (MCAK) is a key regulator of mitotic spindle assembly and dynamics. However, the regulatory mechanisms underlying its depolymerase activity during the cell cycle remain elusive. Here, we showed that PLK1 is a novel regulator of MCAK in mammalian cells. MCAK interacts with PLK1 in vitro and in vivo. The neck and motor domain of MCAK associates with the kinase domain of PLK1. MCAK is a novel substrate of PLK1, and the phosphorylation stimulates its microtubule depolymerization activity of MCAK in vivo. Overexpression of a polo-like kinase 1 phosphomimetic mutant MCAK causes a dramatic increase

in misaligned chromosomes and in multipolar spindles in mitotic cells, whereas overexpression of a nonphosphorylatable MCAK mutant results in aberrant anaphase with sister chromatid bridges, suggesting that precise regulation

of the MCAK activity BLZ945 in vivo by PLK1 phosphorylation is critical for proper microtubule dynamics and essential for the faithful chromosome segregation. We reasoned that dynamic regulation of MCAK phosphorylation by PLK1 is required to orchestrate faithful cell division, whereas the high levels of PLK1 and MCAK activities seen in cancer Navitoclax mouse cells may account for a mechanism underlying the pathogenesis of genomic instability.”
“The purpose of this study is to consider the effect of polydispersity in triblock copolymer on its order-to-disorder phase transition and morphology, by supplementing continuous description of polymer indices carried out using random phase approximation by a discrete sampling of the polydispersity performed at the mesoscale level. Both methods confirm that the increase in polydispersity involves a greater stability of the ordered state, in agreement with published data on the effect of polydispersity on diblock copolymers. The morphology simulations reveal that longer blocks are phase segregated and shorter blocks are more or less uniformly distributed Silmitasertib concentration throughout the polymer. It is proposed that avoiding very short polymer chains will increase the degree of phase segregation, thus improve the mechanical and conductive properties

of proton exchange membrane more efficiently.”
“We demonstrate 260 GHz (lambda = 1.15 mm) near field imaging using a conical Teflon probe whose tip protrudes through an aperture in a tapered aluminum holder. The imaging system is based on a quasioptical millimeter wave vector network analyzer. We present a variety of different imaging examples of dielectrics and metals, in both reflection and transmission modes, as well as an analysis of interesting diffraction and scattering effects observed in some of the images. The probe has an approximate tip diameter of 0.17 mm and an aperture size of about 1 mm. We observe horizontal resolution ranging from 0.2-0.5 mm depending on the sample being imaged. (C) 2009 American Institute of Physics. [DOI: 10.1063/1.

The statistically significant differences were found at the locus

The statistically significant differences were found at the locus EGF: sows with BB genotype showed higher number of piglets born alive (NBA) and higher number of alive piglets on 21st day (N21) (P < 0.01 and P < 0.05) when compared to the other genotypes. At the LIF1 locus sows of AA genotype had higher number of NBA (P < 0.01) when compared to BB genotype, and at the LIF3 sows of BB had PD173074 mouse better results for this trait when compared to AA and AB genotype (P < 0.01 and P < 0.05). In our study, we obtained also statistically significant results for association between AREG gene and

reproductive traits. In parities 2 to 4, statistically significant differences were observed between sows of A1A1 and A1A2 genotype of the AREG gene for NBA (P < 0.05) and between AA and BB genotypes of the LIF1 gene for NBA and N21 (P < 0.01 and P < 0.05). (C)

2013 Elsevier B.V. All rights reserved.”
“Ventricular assist devices are 432 frequently used to bridge pediatric patients to cardiac transplantation; however, experience in single ventricle patients with aortopulmonary shunts remains limited. This case report addresses the challenge of balancing pulmonary and systemic circulation with a focus on the role of continuous versus pulsatile ventricular assist device support.”
“Objective: To develop recommendations for child unintentional injury prevention by comparing New Zealand’s child unintentional injury mortality and injury prevention policies with those of European countries. BI 6727 nmr Methods: Unintentional child injury death rates based on external cause of injury were calculated and ranked. NZ’s score for each of the 12 domains CCI-779 in vivo (based on external causes of injury) from the New Zealand Child and Adolescent Report Card was compared to European scoring. Policy priorities are identified by domains where mortality makes up a high proportion of overall child unintentional injury mortality (high burden of injury) and where report card score for that domain

is low in comparison to other countries (under-utilisation of effective interventions). Results: Death as a motor vehicle occupant accounts for 49% of all child unintentional injury deaths, followed by pedestrian (10%) and drowning deaths (8%). The overall score for the 12 policy domains of the NZ Report Card ranks NZ as 15(th) among the 25 European countries. There are important policy and legislative actions which NZ has not implemented. Conclusions: A number of evidence-based injury prevention policy and legislative actions are available that could target areas of greatest childhood injury mortality in NZ. Implications: A set of injury prevention policy and legislation priorities are presented which, if implemented, would result in a significant reduction in the injury mortality and morbidity rates of NZ children.”
“Traditional colorimetric protein assays such as Biuret, Lowry, and modified Lowry (U-1988) are unsuitable for colored biological samples.

Because of combined clopidogrel and aspirin resistance and to uns

Because of combined clopidogrel and aspirin resistance and to unsuccessful PCI treatment, a single coronary artery bypass graft (CABG) was planned. Awaiting surgery, 3 days later, the fourth ST occurred. It is angiographically confirmed and thus, CABG was

performed. After CABG, in Sotrastaurin chronic treatment with aspirin (300 mg/die) and ticlopidine (500 mg/die), no bleeding complications occurred and the patient did not experience recurrent ischemia (2 years follow-up). A better platelet inhibition by ticlopidine than that obtained by clopidogrel was observed. Our case report remarks the importance to identify these poor responder patients as the treatment can be tailored with alternative therapeutic options (ticlopidine, prasugrel, warfarin) and/or different revascularization strategies (CABG).”
“Background: 4 Near-infrared spectroscopy estimates soft-tissue oxygenation approximately 2 to 3 cm below the skin. The purpose of the present study was to evaluate muscle oxygenation in the setting of an acute compartment syndrome of the leg and to determine if near-infrared spectroscopy SRT2104 cell line is capable of detecting perfusion deficits.\n\nMethods: Fourteen patients with unilateral lower extremity trauma

were enrolled after the diagnosis of an acute compartment syndrome was made clinically and confirmed with intracompartmental pressure measurements. Lower extremity muscle compartments were evaluated with near-infrared spectroscopy, and near-infrared spectroscopy values of the uninjured, contralateral leg of each patient were used as learn more internal reference values. The compartment perfusion gradient was calculated as the diastolic blood pressure minus the intracompartmental pressure.\n\nResults: Intracompartmental pressures ranged from 21 to 176 mm Hg (mean, 79 mm Hg) and exceeded 30 mm Hg in all compartments but two (both in the same patient). Thirty-eight compartments had a perfusion gradient of 510 mm Hg (indicating ischemia). Among ischemic compartments, near-infrared

spectroscopy values in the anterior, lateral, deep posterior, and superficial posterior compartments of the injured limbs were decreased by an average 10.1%, 10.1%, 9.4%, and 16.3% in comparison with the corresponding compartments of the uninjured leg. Differences in near-infrared spectroscopy values (the near-infrared spectroscopy value for the injured leg minus the near-infrared spectroscopy value for the uninjured leg) were positively correlated with compartment perfusion gradient within each compartment (r = 0.82, 0.65, 0.67, and 0.62, for the anterior, lateral, deep posterior, and superficial posterior compartments, respectively; p < 0.05 for all).\n\nConclusions: Normalized near-infrared spectroscopy values decrease significantly with decreasing lower limb perfusion pressures. Near-infrared spectroscopy may be capable of differentiating between injured patients with and without an acute compartment syndrome.

Because of the significant perioperative risk, the demanding oper

Because of the significant perioperative risk, the demanding operative management and the rarity of this tumor, patients with HC should be referred to tertiary surgical centers.”
“Brain tumors are the leading cause of death and disability from childhood disease in developed countries. Pediatric posterior fossa tumors are often effectively controlled with a combination

of surgery, radiation, and chemotherapy, depending on tumor type. White matter injury following resection of tumor and radiation treatment is associated with cognitive declines, including working memory deficits. We investigated how brain injury following treatment for posterior fossa tumors results in deficits in working memory. We used diffusion tensor imaging and probabilistic tractography to examine the structural integrity of cerebello-thalamo-cerebral tracts in patients and healthy children. We also compared working memory outcome in patients versus controls, and related GSK J4 ic50 this function to integrity of cerebello-thalamo-cerebral tracts. Bilateral cerebello-thalamo-cerebral tracts were delineated in all participants. Patients treated with a combination of surgery and radiation had selleck compound lower mean anisotropy and higher mean radial diffusivity within the cerebellar regions of the cerebello-thalamo-cerebral tract compared to patients treated with surgery

only and healthy controls. Poorer working memory scores were observed for the cranial radiation group

relative to controls. Reduced anisotropy and higher radial diffusivity within the entire cerebello-thalamo-cerebral pathway predicted lower working memory. Our finding that working memory function is related to the integrity of cerebello-thalamo-cerebral connections is a novel contribution to the understanding of cerebral-cerebellar communication. Identifying differences in the structural integrity of white matter for specific pathways is an essential step in attempting to learn more localize the effects of posterior fossa tumors and their treatment methods. (C) 2011 Elsevier Inc. All rights reserved.”
“Topographic modulation of tissue response is an important consideration in the design and manufacture of a biomaterial. In developing new tissue therapies for skin, all levels of architecture, including the nanoscale need to be considered. Here we show that keratinocyte phenotype is affected by nanoscale changes in topography with cell morphology, proliferation, and migration influenced by the pore size in anodic aluminum oxide membranes. A membrane with a pore size of 300 nm, which enhanced cell phenotype in vitro, was used as a dressing to cover a partial thickness burn injury in the pig. Wounds dressed with the membrane showed evidence of advanced healing with significantly less organizing granulation tissue and more mature epidermal layers than control wounds dressed with a standard burns dressing.

The average follow-up was 30 1 months The average period from th

The average follow-up was 30.1 months. The average period from the original arthroplasty to the arthrodesis was 49.8 months. Results: Thirteen of the see more 17 ankles were considered radiographically healed after the first attempt in an average time of 3.7 months and 3 after repeat arthrodesis.

Bone grafts were used in 16 patients. The median postoperative AOFAS score was 74.5. The mean Meary angle of the hindfoot was 5 degrees of valgus. Conclusion: Tibiotalar and tibiotalocalcaneal arthrodeses were effective salvage procedures for failed TAR. Massive cancellous allografts were a good alternative to compensate for the large bone defect after removal of the prosthesis and to preserve the leg length. Level of Evidence: Level IV, retrospective case series.”
“SETTING: The risk of pulmonary tuberculosis (PTB) may increase with increased use of anti-tumour necrosis factor (TNF) treatment in inflammatory arthritis. OBJECTIVE: To evaluate the impact of anti-TNF treatment on radiological manifestations of PTB. METHOD: Between January 2007 and December 2012, the chest radiographs (CXRs) of 23 consecutive patients with newly diagnosed PTB who underwent anti-TNF treatment were studied. Chest computed Selleckchem Dibutyryl-cAMP tomography (CT) images were available for 14. To compare the radiological features

of PTB, the CXRs of 46 immunocompetent PTB patients with similar demographics were studied as controls, of whom 34 underwent chest CT. Two radiologists and one chest physician reviewed the chest images independently. RESULTS: Compared with the controls, fibronodular

lesions were less common on Crenigacestat chemical structure CXR in the anti-TNF group (P smaller than 0.001). In contrast, lymphadenopathy (P smaller than 0.001), pleural effusion (P = 0.015) and pericardial effusion (P = 0.02) were more common, while tree-in-bud appearance (P = 0.017) was less commonly depicted on chest CT in the anti-TNF group. Although there was no significant difference in zonal predilection and laterality of the lesions between the two groups, diffuse lesions (P = 0.004) on chest CT scans were more frequent in the anti-TNF group. CONCLUSION: Unusual presentations of PTB were more common in the CXRs and/or CT scans of patients who underwent anti-TNF treatment.”
“Following injury, platelets rapidly interact with the exposed extracellular matrix (ECM) of the vessel wall and the surrounding tissues. Hyaluronan (HA) is a major glycosaminoglycan component of the ECM and plays a significant role in regulating inflammation. We have recently reported that human platelets degrade HA from the surfaces of activated endothelial cells into fragments capable of inducing immune responses by monocytes.

As expected, the initial burst is reduced to 55%, followed by a s

As expected, the initial burst is reduced to 55%, followed by a steady release about 20% from day 1 to day 5 and a slower release of the retained 25% after day 6, resulting in bacterial inhibition zone diameter of 30 mm which can last for more than a month in antibacterial tests, compared with the coated specimen without HA gradually loosing inhibition zone after 21 days. Besides, the cell culture indicates that the vancomycin-chitosan/HA composite coated has enhanced

the proliferation, the differentiation and the mineralization this website of the osteoblast-like cell. In general, it is helpful for the osteointegration on permanent implants. Consistently, it effectively provides the prophylaxis and therapy of osteomyelitis according to the results of the rabbit infection animal model. (C) 2013 Elsevier B.V. All rights reserved.”
“Resorption Z-DEVD-FMK cell line of the alveolar ridge may lead to ridge deformities that make dental implant placement difficult or impossible. Augmentation of the alveolar ridge may restore appropriate ridge form to allow implant placement. Forty-four patients with edentulous spaces completed this multicenter prospective trial to clinically and radiographically evaluate the efficacy of a bovine pericardium membrane and a particulate mineralized cancellous bone allograft in promoting lateral ridge augmentation. Overall,

38 of 44 patients (86.4%) were able to receive dental implants in the appropriate restoratively driven position 6 months after ridge augmentation. The mean gain in clinical ridge width after augmentation was 2.61 mm, while radiographically the mean gain in ridge width was 1.65 mm at a level 3 mm apical to the bony crest and 1.93 mm at a level 6 mm apical to the crest. On average, approximately 50% of the graft material added horizontally

during surgery was displaced or resorbed during healing. Histomorphometric evaluation of cores taken from the augmented ridge at 6 months revealed that approximately 58% of the tissue volume was vital bone, with 12% residual allograft particles and 30% nonmineralized tissue.”
“This paper presents the results of fatigue Smoothened Agonist strength estimation depending on the variable orientation of the critical plane for proportional and non-proportional bending and torsion with regard to specimens made of aluminium alloy 2017A. The algorithm applied for the fatigue strength evaluation is based on the Carpinteri-Spagnoli proposal and its subsequent modifications in accordance with the ideas developed by the authors. The objective of this paper is to search for a model which offers the best possible results of fatigue strength estimation for materials with properties which are intermediate between elastic-brittle and elastic-plastic, such as aluminium alloys, and a further insight into them with regard to their fatigue strength. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.

All rights reserved “
“Objectives: To investigate auditory d

All rights reserved.”
“Objectives: To investigate auditory dysfunction in patients with tinnitus and normal hearing thresholds using two sensitive audiological measures. Design: The study was designed to investigate the characteristics of Audioscan and DPOAE tests in tinnitus patients with normal hearing thresholds. Poziotinib concentration Audioscan and DPOAE notches were analysed and compared. All tests were performed in a sound-treated chamber or in a sound-treated

room. Study sample: Forty-five tinnitus patients with normal hearing thresholds were examined following a written clinical protocol. Results: The averaged hearing levels obtained from tinnitus participants were significantly worse at high frequencies than those derived from the normative data. There was a significantly higher prevalence of Audioscan and DPOAE

notches, whose central frequencies matched tinnitus frequencies in the mid-frequency regions, but not in the low-and high-frequency regions. A significant correlation was found between the centre frequencies of the Audioscan notches and the DPOAE notches from 500 to 4000 Hz. Conclusion: Tinnitus in different frequency regions may be associated with different underlying mechanisms of tinnitus generation. Some negative results on the Audioscan and DPOAE notches matching tinnitus pitches may be due to a limited set of discrete frequencies used for the tinnitus pitch matching test.”
“Structural, spectroscopic and optical properties of lead tellurite glass ceramics doped with variable amounts of erbium ions and codoped with fixed amounts of silver (Ag2O or Ag metallic nanoparticles, AgNPs) have been studied. The investigation of the samples was performed by X-ray diffraction (XRD), Fourier transform infrared (FTIR), diffuse reflectance ultraviolet visible (DR-UV-vis) and photoluminescence (PL) spectroscopies. The average unit-cell parameter, crystallite size and the quantitative ratio of the crystallographic phases in the A-1210477 nmr samples were evaluated from the XRD data. FTIR spectroscopy data show that

the glass ceramics network mainly consists of TeO3 and TeO4 structural units. The increase of the Er2O3 content determines a TeO3 – bigger than TeO4 conversion but the process is also influenced by the nature of the codopant (Ag2O or AgNPs) present in the samples. The optical band gap (E-g(opt)) values, determined from the DR-UV vis spectra, decrease with increasing the erbium content of the samples. Photoluminescence spectroscopy measurements evidenced the important peaks located at around 430 nm and 708 nm due to the Pb2+ ions and at 525 and 547 nm due to the Er3+ ions present in the studied samples. The Er3+ :Ag codoping was found to produce antagonistic effects (quenching/enhancement) of the luminescence emission of Er3+ ions in the studied samples. (C) 2014 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

In this study, two types of relays are considered Conventional a

In this study, two types of relays are considered. Conventional amplify and 3 forward relays Entinostat supplier in which all relays amplify their received signal and forward it to the destination in a round-robin fashion. In addition, decode and forward relays in which the relays that correctly detect the source signal will forward the corresponding fading gain to the destination in pre-determined orthogonal time slots are studied. The optimum decoder for both communication systems is derived and performance analysis are conducted. The exact average bit error probability (ABEP) over Rayleigh fading channels is obtained in closed-form for a source equipped

with two transmit antennas and arbitrary number of relays. Furthermore, simple and general asymptotic expression for the ABEP is derived and analyzed. Numerical results are also provided, sustained by simulations which corroborate the exactness selleck kinase inhibitor of the theoretical analysis. It is shown that both schemes perform nearly the same and the advantages and disadvantages

of each are discussed.”
“Development of assays to screen milk for economically motivated adulteration with foreign proteins has been stalled since 2008 due to strong international reactions to the melamine poisoning incident in China and the surveillance emphasis placed on low molecular weight nitrogen-rich adulterants. New screening assays are still needed to detect high molecular weight foreign protein adulterants and characterize this understudied potential risk. A rapid turbidimetric method was developed to screen milk powder for adulteration with insoluble plant proteins. Milk powder samples spiked with 0.03-3% by weight of soy, pea, rice, and wheat protein isolates were

extracted in 96-well plates, and resuspended pellet solution absorbance was measured. Limits of detection ranged from 100 to 200 mu g, or 0.1-0.2% of the sample weight, and adulterant pellets were visually apparent even at similar to 0.1%. Extraction recoveries ranged from 25 to 100%. Assay sensitivity and simplicity indicate that it would be ideally suitable to rapidly screen milk samples in resource RG-7388 in vivo poor environments where adulteration with plant protein is suspected.”
“BackgroundAlopecia areata (AA) is a common dermatological problem that manifests as sudden loss of hair without any inflammation or scarring. Various cytokines are implicated in the pathogenesis of this disease. Macrophage migration inhibitory factor (MIF) is located at an upstream position in the events leading to the possible dysregulated immuno-inflammatory responses, and the high level of this cytokine in AA may suggest a role of MIF in the pathogenesis of AA. MethodsThis case-control study was carried out on 31 AA patients with different grades of severity and 15 apparently healthy subjects. Serum MIF level was measured by ELISA, and was correlated with the clinical severity of the disease using SALT (severity of alopecia tool) scoring system.

The HCV viral load was determined using an in-house real-time PCR

The HCV viral load was determined using an in-house real-time PCR assay targeting the 5′-NCR. Results: HCV subtypes 1b, 1a, and 3a were found in 45.5%, 32.0%, and 18.0% of the donors, respectively. The mean viral load of genotype 1 was significantly higher than that of the genotype 3 isolates. Subtype 1a was more frequent among young donors and 3a was more frequent among older donors. Protease 3 inhibitor-resistant variants were detected in 12.8% of the sequenced samples belonging to genotype 1, and a higher frequency was observed among subtype 1a (20%) in comparison to 1b (8%). There was no difference in the prevalence of HCV risk factors among the genotypes or drug-resistant

variants. Conclusions: We found a predominance of subtype 1b, with an increase in the frequency of subtype 1a, in young subjects. Mutations conferring resistance to NS3 inhibitors were frequent in treatment-naive blood donors, particularly those buy PD173074 infected with subtype 1a. These variants were detected in the major viral population of HCV quasispecies, have replicative capacities comparable to nonresistant strains, and could be important for predicting

the response to antiviral triple therapy.”
“Objectives: The aim of this study was to compare the long-term results of 2 surgical strategies for patients with bilobar colorectal liver metastases (bCRLM). Background: Two-stage hepatectomy is the surgical strategy mostly chosen for treating extensive BCLM with the pitfall of dropout after the first stage. One-stage strategy combining limited resections and radiofrequency ablation could be proposed find more as an option Y-27632 order in this population. Patients and Methods: Between 2000 and 2010, 272 patients were consecutively operated in 2 expert centers practicing 1- or 2-stage hepatectomy for bCRLM. A case-match study (1: 1) was conducted using number and size of nodules, synchronous presentation, primary node status, and extrahepatic disease

as matching variables to compare overall survival (OS) and disease-free survival (DFS). The analysis was performed in intention to treat, including patients who did not undergo the second stage. Results: In the case-match analysis (156 matched patients), median OS and DFSdid not differ significantly between patients in 1- and 2-stage hepatectomy, respectively: 37.2 and 34.5 months (P = 0.6), 9.4 and 7.5 months (P = 0.25). Multivariate analysis confirmed the absence of impact of strategy on OS and DFS. Primary advanced T stage and synchronous presentation were predictors of poor OS (HR = 3.67 and 1.92); CEA more than 200 ng/mL, absence of postoperative chemotherapy, and extrahepatic disease were predictive of recurrence (HR = 2.77, 1.85 and 1.69, respectively). Conclusions: This first case-match study demonstrates that on an intention-to-treat analysis 1- and 2-stage hepatectomy in patients with bCRLM achieve comparable OS and DFS, despite the high dropout of the 2-stage strategy.