The most abundant component in the essential oils of the flowers

The most abundant component in the essential oils of the flowers was (2E,6E)-methyl farnesoate, whereas hexadecanoic acid

was the most abundant essential oil component in the stem bark. The leaf essential oils showed seasonal variation in their chemical composition, with bicyclogermacrene and (2E,6E)-methyl farnesoate as the major chemical components. Forty-four constituents were identified, and only nine compounds were found in all of the samples. Sesquiterpenes were mainly produced in the flowers and leaves. The PCA showed a positive correlation between the oxygenated OICR-9429 in vivo sesquiterpenes and the foliar nutrients Cu and P. Significant statistical correlations were verified between the climatic data, foliar nutrients and essential oil compositions.”
“A screening was undertaken of the native flora of Chile excluding Pteridophyta, Cactaceae and Poaceae the study included 396 species. Alkaloids were found in 189 species, the highest concentration being 3 mg/g Selleckchem Rabusertib dry tissue. A table was produced listing all

species collected and specifying the subset containing alkaloids, the species with a particularly high alkaloid content and also the endemic species within this latter set. Alkaloid concentration as well as presence or absence data in different vegetational subregions correlated with mean collection latitude, under the limitations imposed by the relationship between alkaloid concentration and species lineage. In practical terms, the results

indicate that in studies aimed at identifying alkaloids in the native flora of Chile, plant collections should take into consideration the promising species listed in the table and take place at the lowest possible latitude if the chance of finding alkaloids and the yield of isolated alkaloids are to be maximized.”
“Atherosclerosis is the main cause of coronary artery aneurysm, however they can be observed in connective tissue diseases such as systemic lupus erythematosus and vasculitis. Kawasaki’s disease and polyarteritis nodosa (PAN) are the systemic vasculitis that more often present coronary artery aneurysms. There are descriptions in the literature that small vessel vasculitis such as microscopic polyangiitis and PAN MAPK inhibitor could develop coronary artery aneurysm, which are infrequent in other ANCA-associated vasculitis. Here, we report the case of a 25-year-old man who developed an extensive anterior myocardial infarct. The coronary angiogram showed coronary artery aneurysms, on laboratory ANCA C positivity with elevated levels of anti-proteinase 3 antibodies were present. He was treated with high doses of corticosteroids and cyclophosphamide with resolution of the aneurysms. (C) 2010 Societe francaise de rhumatologie. Published by Elsevier Masson SAS. All rights reserved.”
“This paper studied the adequacy of the World and China lithium resources, considering the most promising uses in the future, involving nuclear fusion and electric-vehicles.

The factors controlling vascular patterning are not yet completel

The factors controlling vascular patterning are not yet completely understood. Recent studies have highlighted the anatomic and structural similarities between blood vessels and nerves. The 2 networks are often aligned, with nerve fibers and blood vessels following parallel routes. Furthermore, both systems require precise control over their guidance and growth. Several molecules with attractive and

Selleckchem Screening Library repulsive properties have been found to modulate the proper guidance of both nerves and blood vessels. These include the Semaphorins, the Slits, and the Netrins and their receptors. In this review, we describe the molecular mechanisms by which blood vessels and axons achieve proper path finding and the molecular cues that are involved in their guidance. (Circ Res. 2009;104:428-441.)”
“Metabolic syndrome (MS) is associated with an increased risk of coronary heart disease, stroke, and cardiovascular mortality; but its effect on patients undergoing cardiac revascularization is still unclear. Robust evidence demonstrates that diabetes mellitus and insulin resistance are among the main risk factors for restenosis in patients requiring percutaneous myocardial revascularization. The recent advent of drug-eluting stents (DESs) has significantly

reduced the incidence of restenosis compared with bare-metal stents, both in nondiabetic and in diabetic patients. The aim of the study was to evaluate the effect of MS on the risk of binary restenosis in DES implant recipients. One hundred eighty-nine recipients of successful DES implants ML323 manufacturer performed between January and March 2005 for stable coronary artery disease

underwent 1-year clinical and angiographic follow-up. Body mass index (BMI), blood pressure, fasting blood glucose, and lipid profile were determined. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III criteria, with the waist criterion being substituted by a BMI >= 28.8 kg/m(2). Metabolic and anthropometric information for MS diagnosis was available for 148 of 189 patients; 87 of 148 patients (58%) had MS. Patients with MS had higher BMI (28.4 +/- 3.8 VX-680 vs 26 +/- 2.7 kg/m(2), p <.0001), systolic blood pressure (133 +/- 14 vs 124 +/- 14 mm Hg, P = .0004), and fasting glucose (113 +/- 37 vs 92 +/- 17 mg/dL, P < .0001). They also had higher serum triglycerides (154 +/- 94 vs 113 +/- 43, P = .0018) and lower high-density lipoprotein cholesterol levels (39 +/- 9 vs 46 +/- 10, P <.0001). Rates of restenosis (10.5% vs 8.1%, P = not significant [NS]), target vessel revascularization (10.5% vs 11.3%, P = NS), and major adverse cardiac events (11.6% vs 14.5%, P = NS) were not significantly different in patients with MS compared with those without MS, nor was any association found between increased end point risk and presence of MS.

Results: H pylori was eradicated in 21 (46 6%) pts in the LAC gr

Results: H. pylori was eradicated in 21 (46.6%) pts in the LAC group, while it was eradicated in 29 (64.4%) pts in the LACE

group. There was no statistical difference between the two groups (p=0.13). Conclusion: The eradication rate of H. pylori in children while using the LAC regimen has decreased in the last years. The LACE regimen has been associated with an increased eradication rate but can reach to statistically significance. Further studies SB203580 with larger cohorts are needed to examine the success of the LACE regimen for H. pylori eradication.”
“Mesenchymal neoplasms of the vulvovaginal and inguinoscrotal regions are among the most diagnostically challenging specimens in the pathology laboratory owing largely to their unique intersection between general soft tissue tumors and relatively genital-specific mesenchymal tumors. Genital stromal tumors are a unique

subset of soft tissue tumors encountered at this location, and this group includes fibroepithelial stromal polyp, FDA-approved Drug Library molecular weight superficial (cervicovaginal) myofibroblastoma, cellular angiofibroma, mammary-type myofibroblastoma, angiomyofibroblastoma and aggressive angiomyxoma. Aside from the striking morphologic and immunophenotypic similarity that is seen with these entities, there is evidence that a subset of genital stromal tumors may be linked genetically. This review will focus on simplifying this group of tumors and provide the pathologist or dermatopathologist with practical management information. Smooth muscle tumors of the external genitalia will also be discussed.”
“Treatment with the chemotherapeutic agent oxaliplatin produces a robust painful neuropathy similar to various other neuropathic find more conditions which result in loss of nerve fibers innervating the skin. This loss of intraepidermal nerve fibers (IENFs) appears to play an important role in neuropathy, but has yet to be investigated in oxaliplatin-induced

neuropathic pain. For this study, mechanical hyperalgesia and IENF density were measured in rats receiving oxaliplatin, given at a dosage of 2 mg/kg every other day for four injections. The immunomodulatory agent minocycline (25 mg/kg) was also administered and was given 24 h prior to the first dose of oxaliplatin and continued throughout oxaliplatin treatment. Immunohistochemistry using the pan-neuronal marker PGP9.5 was used to investigate IENF densities in hind paw skin on Day 15 and Day 30. The results show that a robust mechanical sensitivity developed in oxaliplatin treated animals, as did a pronounced decrease in epidermal nerve fibers, and these outcomes were effectively prevented by minocycline treatment. This is the first study to show changes in IENF density in oxaliplatin treated animals, and confirm not only a relationship between IENF loss and hypersensitivity but also prevention of both with minocycline treatment (C) 2011 Elsevier Inc. All rights reserved.

Fifty-eight patients

\n\nFifty-eight patients PSI-7977 underwent complete radiographic staging. Nineteen (33%) had ulcerated primary tumors. Forty-two patients (73%) presented with clinically negative lymph nodes that were positive on sentinel lymph node biopsy. Lymph

node involvement was classified as N1a in 54%, N2a in 19%, N2b in 3%, and N3 in 22% of patients. Among 204 staging examinations in 58 patients, 52 (25%) were initially reported as positive. Three percent of all examinations proved truly positive; 23% were falsely positive. Analyzed per patient, in 37 (64%) of 58 patients, at least one examination was initially reported as positive. However, only 3 patients (5%) had a true-positive and 34 (59%) had at least one false-positive report. The positive

reports of the staging scans generated 45 additional examinations (0.78 per patient).\n\nRadiographic staging in asymptomatic patients with stage III melanoma detects a low number of patients with unsuspected systemic disease. The ratio of falsely to truly positive is approximately 11:1. Radiographic screening should only be considered in patients with high-risk prognostic features or symptoms, or in the context of clinical trials.”
“The asymmetric unit of the title compound, C(9)H(8)Br(3)NO, contains two independent molecules. The conformation of the N-H bond is anti to the 3-methyl substituent in the benzene ring in each molecule. The structure shows both intramolecular N-H center dot center dot center dot Br and intermolecular N-H center dot center dot SRT1720 solubility dmso center dot O hydrogen bonding, the latter leading to the formation of helical supramolecular chains along the b axis.”
“Purpose: This study compared patients who underwent cardiac resynchronization therapy (CRT) by high-amplitude left ventricular (LV) pacing with those who underwent CRT by standard LV pacing.\n\nMethods: We included 32 CRT patients with ejection fraction (EF) <= 35%, QRS time >= 120 ms, and New York Heart Association (NYHA) class III/IV symptoms of heart failure despite optimal medical treatment. These patients were evaluated clinically and echocardiographically before, three

and six months after CRT. At the 3rd month, the LV pulse amplitude selleck value was set high at 5 volt for 16 patients [high-amplitude Group (HAG)], while for the other 16 patients, it was reduced to at least twice the threshold value at <= 2.5 volt [low-amplitude group (LAG)].\n\nResults: Clinical and echocardiographic response rates of HAG and LAG after CRT were similar in the 3rd and 6th month. In both groups, increase in LVEF and decrease in LV ESV in the 3rd and 6th month were statistically significant compared to those before CRT, and NYHA class and end-diastolic volume (EDV) was significantly reduced in the 6th month compared to those before CRT. However, NHYA class and EDV continued to reduce significantly in HAG from the 3rd to the 6th month (P<0.05), while the decrease in LAG was not significant (P>0.05).

We have previously argued that unrepaired

G:T mismatches

We have previously argued that unrepaired

G:T mismatches from spontaneous deamination of 5-methylcytosine at CpG sites could be converted to apparent in vivo mutations in the bacterial recovery systems because of rapid, random, mismatch repair in Escherichia coli. In this study, we have measured mutation frequencies in spleen of male mice induced by N-ethyl-N-nitrosourea (ENU) using the Phi X174 transgene, which is not subject to mismatch repair in E.coli, using single-burst analysis, a unique method to identify in vivo mutation. In order to compare our results to those using the lacI and cII transgenes, we converted all mutant frequencies to base pair Autophagy inhibitor in vitro substitution (bps) mutation frequencies per nucleotide based on mutant spectra from this study and published literature. We found this frequency in control spleen to be similar for lacI (3.8 +/- 0.7 x 10(-8)) and Phi X174 (3.1 +/- 1.2 x 10(-8)) at 6 weeks of age. We found a strong age dependence for spontaneous lacI mutation that extrapolated to a value at conception (1.8 +/- 0.9 x 10(-8)) that was not significantly different from the human germ line bps mutation frequency per nucleotide of 1.7 +/- 0.2 x 10(-8). These two transgenes provided similar

mutational responses to 40 mg/kg ENU, 7- to 9-fold. In contrast, the cII target gene in the same tissue produces both spontaneous and induced mutation frequencies similar to 10 times higher, for unknown reasons. We conclude that the spontaneous mutant frequencies measured by the lacI and Phi X174 transgenes in this moderately dividing tissue accurately measure in vivo mutation frequencies at early ages. For these two transgenes, Wnt activation seemingly high mutant frequencies may reflect the expected accumulation of somatic mutation with age.”
“Extracellular purine and pyrimidine

compounds induce a multiplicity of cellular signal pathways that can induce multiple trophic functions. They interact with other low molecular weight messengers, growth factors, and extracellular matrix components. An increasing number of studies now provide evidence for a role of purinergic signaling in neural development, including progenitor cell proliferation, cell migration, neuronal and glial maturation and differentiation, and cell death and survival. This brief overview highlights recent developments supporting a contribution of purinergic signaling to embryonic and adult neurogenesis. (C) 2011 Elsevier Ltd. All rights reserved.”
“We report a case of bilateral ulna stress fractures following bilateral femoral fractures associated with long-term bisphosphonate use. The patient is an 84-year-old woman receiving 15 years of bisphosphonate therapy.

“Purpose: To report the clinical efficacy and intermediate

“Purpose: To report the clinical efficacy and intermediate-term functional outcome after laparoscopic anatrophic nephrolithotomy (LAN) as an alternative treatment modality for complete staghorn renal stone.\n\nPatients and Methods: The demographic and perioperative parameters as well as the intermediate outcome of 10 adults (9 men) who underwent transperitoneal LAN for complete staghorn renal stone were analyzed. Functional imaging studies consisted of intravenous urography (IVU) and technetium-99 dimercaptosuccinic acid scintigraphy (99Tc-DMSA) renal scan done before the operation and at the last follow-up visit.\n\nResults: Mean age of patients was 48.7

years (range 37-64 years). Mean stone size was 67.3 mm (50-90 mm). Mean

Prexasertib operative time was 192 minutes (110-240 min), and mean warm ischemia time was 32.8 minutes (15-40 min). A few hours after laparoscopy, one patient underwent splenectomy because TGF-beta inhibitor of significant hemorrhage from a splenic laceration (grade IIIb complication). During the follow-up period early after the operation, we detected an 8-mm lower caliceal stone and a 25-mm midcaliceal stone in one patient each (stone-free rate: 80%). After a mean follow-up of 11.9 months (6-19 mos), 85.5% of corresponding renal unit function was preserved; however, there was a significant mean decrease in 99Tc-DMSA uptake from 48.4% +/- 8.83 before surgery to 41.4% +/- 13.98 afterward (-7% +/- 6.53; P = 0.008). Nevertheless, renal units were completely functional at follow-up IVU with a significant improvement in obstruction in all patients.\n\nConclusion: LAN is an alternative minimally invasive approach for

one-session management of patients with complete staghorn renal stone. It offers an acceptable rate of stone clearance and operative complications but does incur a minimal loss of function in the affected kidney.”
“Transcranial direct current stimulation (tDCS) was recently proposed for the treatment of epilepsy. However, the electrode arrangement for this case is debated. This paper analyzes the influence of the position of the anodal electrode on the electric field in the brain. The simulation shows that moving the anode from scalp to shoulder does influence the electric field not only in the cortex, but also in deeper brain regions. The electric field decreases dramatically in the brain area without epileptiform activity.”
“Objectives: The aim of this study is to perform a comparative costs analysis of radical retropubic prostatectomy (RRP) and robotic-assisted laparoscopic prostatectomy (RAP) for clinically localized prostate cancer and to determine whether to expand the use of RAP or to continue with conventional RRP in Teaching Hospital San Giovanni Battista Turin Italy.\n\nMethods: A cohort study was carried out comprising consecutive patients undergoing radical prostatectomy.

Recent studies have demonstrated activity-dependent regulation of

Recent studies have demonstrated activity-dependent regulation of the AIS, including homeostatic changes in AIS length, membrane excitability, and the localization of voltage-gated Na+ channels. The neurodevelopmental disorder Angelman syndrome (AS) is usually caused by the deletion of small portions of the maternal copy of chromosome 15, which includes the UBE3A gene. A mouse model of AS has been generated and these mice exhibit multiple neurological abnormalities similar to those observed in humans. We examined intrinsic properties of pyramidal neurons in hippocampal area CA1 from AS model mice and observed alterations in resting membrane potential, threshold potential,

and action potential amplitude. The altered intrinsic properties in the AS mice were Mizoribine DNA Damage inhibitor correlated with significant increases in the expression of the alpha 1 subunit of Na/K-ATPase (alpha 1-NaKA), the Na+ channel NaV1.6, and the AIS anchoring protein ankyrin-G, as well as an increase in length of the AIS. These findings are the first evidence for pathology of intrinsic membrane properties and AIS-specific changes in AS, a neurodevelopmental disorder associated with autism.”
“A genetic linkage map of tetraploid wheat was constructed based on a cross CX-6258 ic50 between durum wheat [Triticum turgidum ssp. durum (Desf.) MacKey] cultivar Langdon

and wild emmer wheat [T. turgidum ssp. dicoccoides (Korn.) Thell.] accession G18-16. One hundred and fifty-two single-seed descent derived F(6) recombinant inbred lines (RILs) were analyzed with a total of 690 loci, including 197 microsatellite and 493 DArT markers. Linkage analysis defined 14 linkage groups. Most markers were mapped to the B-genome (60%), with an average of 57 markers per chromosome and the remaining 40% mapped to the A-genome, with an average of 39 markers per chromosome. To construct a stabilized (skeleton) map, markers interfering with map stability were removed. The skeleton map consisted

of 307 markers with a total length of 2,317 cM and average distance of 7.5 cM between adjacent markers. The length of individual chromosomes Histone Methyltransf inhibitor ranged between 112 cM for chromosome 4B to 217 cM for chromosome 3B. A fraction (30.1%) of the markers deviated significantly from the expected Mendelian ratios; clusters of loci showing distorted segregation were found on chromosomes 1A, 1BL, 2BS, 3B, and 4B. DArT markers showed high proportion of clustering, which may be indicative of gene-rich regions. Three hundred and fifty-two new DArT markers were mapped for the first time on the current map. This map provides a useful groundwork for further genetic analyses of important quantitative traits, positional cloning, and marker-assisted selection, as well as for genome comparative genomics and genome organization studies in wheat and other cereals.

Here, we subjected iPSCs of swine to a rod photoreceptor differen

Here, we subjected iPSCs of swine to a rod photoreceptor differentiation protocol consisting of floating culture as embryoid bodies followed by differentiation in adherent culture. Real-time PCR and immunostaining of differentiated cells demonstrated loss of expression of the pluripotent genes POU5F1, NANOG, and SOX2 and induction of rod photoreceptor genes RCVRN, NRL, RHO, and ROM1. While these differentiated cells

displayed neuronal morphology, IWR-1-endo supplier culturing on a Matrigel substratum triggered a further morphological change resulting in concentration of rhodopsin (RHO) and rod outer segment-specific membrane protein 1 in outer segment-like projections resembling those on primary cultures of rod photoreceptors. The differentiated cells were transplanted into the subretinal space of pigs treated with iodoacetic acid to eliminate rod photoreceptors. Three weeks after transplantation,

engrafted RHO(+) cells were evident in the outer nuclear layer where photoreceptors normally reside. A portion of these transplanted cells had generated projections resembling outer segments. These results demonstrate that iPSCs of swine can differentiate into photoreceptors in culture, and these cells can integrate into the damaged swine neural retina, thus, laying a foundation for future studies using the pig as a model for retinal stem cell transplantation. STEM CELLS 2011;29:972-980″
“A multisensor system combined with multivariate analysis FLT3 inhibitor is applied for the characterization and classification of white grape juices. The proposed system, known as hybrid electronic tongue, consists of an array of electrochemical microsensors and a colorimetric optofluidic system. A total of 25 white grape juices representing the large variability of vines grown in the Northwest Iberian Peninsula were studied. The data obtained were treated with Principal Component Analysis (PCA) and Soft Independent Modeling Class Analogy (SIMCA). The first tool was used to train the system

with the reference genotypes Albarino, Muscat a Petit Grains Blanc and Palomino and the second to study the feasibility of the hybrid electronic tongue to distinguish between different grape juice varieties. The results show that the three reference genotypes are well differentiated in the PCA model and this can be used to interpolate the rest of varieties and predict their basic characteristics. Besides, using the SIMCA, the system demonstrates high potential for classifying and discriminating grape varieties.”
“Although Paget’s disease of bone (PDB) is the second most common metabolic bone disease, to our knowledge, there is only one quantitative analysis on the histological and especially on the histomorphometric level.

Improved patient ventilator interaction, sufficient unloading of

Improved patient ventilator interaction, sufficient unloading of the respiratory muscles and increased comfort have been recently associated with these ventilator modalities. There are, however, scarce data with regard to outcome improvement, such as length of mechanical ventilation, ICU stay or mortality Ubiquitin inhibitor (commonly accepted targets to demonstrate clinical superiority).\n\nSummary\n\nWithin recent years, a major step forward in the evolution of assisted (effort-adapted) modes

of mechanical ventilation was accomplished. There is growing evidence that supports the physiological concept of closed-loop effort-adapted assisted modes of mechanical ventilation. However, at present, the translation into a clear outcome benefit remains to be proven. In order to fill the knowledge gap

that impedes the broader application, larger randomized controlled trials are urgently needed. However, with clearly proven drawbacks of conventional assisted modes such as pressure support ventilation, it is probably about time to leave these modes introduced decades ago behind.”
“Histoplasmosis and paracoccidioidomycosis are emerging infections in Spain associated with immigration and travelling. In last three decades a total of 128 cases of histoplasmosis have been reported in Spain, 59 in travellers, 63 in immigrants, three associated to drug abuse, two in laboratory workers, and one in a solid organ transplant receptor. In 1969 the first Spanish case

of paracoccidioidomycosis was published and MK 2206 a total of 21 cases have been reported so far. Those patients suffered from the chronic form of the disease with period of latency Nocodazole as long as 50 years. Other endemic mycoses such as blastomycosis, coccidioidomycosis, lobomycosis, pythiosis and sporotrichosis have not increased in frequency. Microbiological cultures of endemic fungi must be handled in facilities which comply with international biosafety regulations and must also be taken into account for cultures from patients with suspected endemic mycosis. (C) 2011 Elsevier Espana, S.L. All rights reserved.”
“The recent development of the analytical techniques offers the unprecedented possibility to study simultaneously concentration of dozens of elements in the same biological matrix sample of 0.5 – 1.0 g (multielement profiles). The first part of this essay entitled “Think globally … An outline of trace elements in health and disease” aims to introduce the reader to the fascinating field of elements, there importance to our nutrition, their essentiality, deficiency, toxicity and bioavailability to the body and their overall role in health and disease, including the genetic metabolic impairments. In the second part of the essay entitled ” … and act locally.

The underlying mechanisms of action, however, remain largely unkn

The underlying mechanisms of action, however, remain largely unknown. Here, using a mouse hepatocellular carcinoma (HCC) model, the authors found an important and previously unidentified effect of metformin Cell Cycle inhibitor with the inhibition of HCC growth through an indirect path mediated by IL-22. The findings also provide evidence for immune-modulatory effects of metformin in HCC. Taken together, these data broaden our current understanding

of the mechanisms of action of metformin in liver cancer treatment.”
“Background Swallowing dysfunction can lead to recurring aspiration and is frequently associated with chronic symptoms such as cough and wheezing in infants. Our objective was to describe the characteristics of infants with swallowing dysfunction, determine if pulmonary function abnormalities are detectable, and if they improve after therapy. Methods We studied 38 infants with a history of

coughing and wheezing who had pulmonary function tests performed within two weeks of their diagnosis of swallowing dysfunction. The raised lung volume rapid thoracoabdominal compression technique was used. After 6 months of therapy, 17 of the infants repeated the tests. Results Initially, 25 had abnormal spirometry, 18 had abnormal plethysmography, and 15 demonstrated bronchodilator responsiveness. Six months later test were repeated for seventeen patients. Ten patients had continued abnormal spirometry, two patients remained normal, Selleckchem VS-6063 three patients’ abnormal spirometry had normalized, and two patients’ previously normal studies became abnormal. Eight of the 17 patients had continued abnormal plethysmography, six had continued normal plethysmography, and three patients’ normal plethysmography became abnormal. After 6 months of treatment, eight patients demonstrated bronchodilator responsiveness, of which five continued to demonstrate bronchodilator responsiveness

and three developed responsiveness. The remainder either continued learn more to be non-bronchodilator responsive (two) or lost responsiveness (three.) The findings of the abnormal tests in most infants tested is complicated by frequent occurrence of other co-morbidities in this population, including gastroesophageal reflux in 23 and passive smoke exposure in 13 of the infants. Conclusions The interpretation of lung function changes is complicated by the frequent association of swallowing dysfunction with gastroesophageal reflux and passive smoke exposure in this population. Six months of medical therapy for swallowing dysfunction/gastroesophageal reflux did not significantly improve pulmonary function in these infants. Long-term studies will be necessary to determine which of these changes persists into adulthood.”
“Here we investigate the utility of a dual-echo Echo-Planar Imaging (DE-EPI) Diffusion Weighted Imaging (DWI) approach to improve lesion conspicuity in pediatric imaging.