To better define the physiological properties of NG2(+) cells, we

To better define the physiological properties of NG2(+) cells, we used transgenic mice that allowed an unbiased sampling of this population and unambiguous identification of cells in discrete states of differentiation. Using acute brain slices prepared from developing and mature mice, we found that NG2(+) cells in diverse brain regions share a core set of physiological properties, including expression of voltage-gated Na(+) (NaV) channels and ionotropic glutamate receptors, and formation of synapses with glutamatergic neurons. Although small amplitude Na(+) spikes could be elicited in some NG2(+) cells during the first postnatal week, they were not capable of generating action potentials. Transition of these

progenitors to the premyelinating stage was accompanied by VX-770 nmr see more the rapid removal of synaptic input, as well as downregulation of AMPA and NMDA receptors and NaV channels. Thus, prior reports of physiological heterogeneity among NG2(+) cells may reflect analysis of cells in later stages of maturation. These results suggest that NG2(+) cells are uniquely positioned within the oligodendrocyte lineage to monitor the firing patterns of surrounding neurons.”
“Cellulase was immobilized on chitosan by the method of covalent binding. The optimum immobilized conditions were as follow: the pH value was 5.0, the glutaraldehyde concentration was 0.015 (w/v) and the formaldehyde concentration was 0.15 (w/v). Both the free

and immobilized cellulase were characterized by determining the pH, temperature, thermal stability and storage stability. The optimum pH of both the free and immobilized cellulase was found as 4. The immobilized cellulase had optimum temperature of 50 degrees C as compared to 40 degrees C in case of free enzyme. The immobilized enzyme showed higher thermal stability than the free cellulase, after 120 min, the YH25448 price activity of immobilized cellulose and the free enzyme retained 86.5 and 61%

respectively. After 11 cycles, the activity of the immobilize enzyme conserved 80.27%. The immobilized enzyme exhibited slightly better storage stability than the free enzyme. The Km and Vm values for the immobilized and free cellulase were 8.1 and 1.84 mg/L and 0.01 and 0.0036 mg/ml/min respectively. Cellulose hydrolysis by immobilized cellulase in the presence of a 88 ionic liquid (IL), 1,3-dimethylimidazolium dimethylphosphate (MMIM-DMP), was investgated. The result showed that the addition of 20% (v/v) MMIM-DMP gave the highest initial rate, which was 1.3 and 13.9 times higher than the hydrolysis rate in citric acid – sodium hydrogen phosphate buffer and in IL, respectively.”
“Animals are known to exhibit ‘personality’; that is, individual differences in behaviour that are consistent across time and/or situations. One axis of personality of particular importance for behavioural ecology is boldness, which can be defined as the tendency of an individual to take risks.

Hereby, the parameter G(max)/T(max) was calculated, derived from

Hereby, the parameter G(max)/T(max) was calculated, derived from the plateau of grey-level intensity (G(max)), divided by the time-to-peak intensity (T(max)). Cardiac magnetic resonance imaging (CMR) deemed as the standard reference

for the estimation of infarct size, transmurality and of the LV-function at 6 months of follow-up. Results: Cut-off values of G(max)/T(max)=5.7/sec and 3.8/sec, respectively, yielded similar accuracy as infarct transmurality for the prediction of follow-up ejection fraction >55% (AUC = 0.86 for STEMI and AUC = 0.90 for NSTEMI, by G(max)/T(max) and AUC = 0.85 for STEMI and AUC = 0.89 for NSTEMI, by infarct transmurality, respectively, P = NS). Both clearly surpassed the predictive value of visual MBG (AUC = check details 0.69 for STEMI and AUC = 0.68 for NSTEMI, P < 0.05). Conclusion: G(max)/T(max) is an easy to acquire but highly valuable surrogate parameter for infarct size, which yields equally high accuracy with infarct transmurality and favorably compares with visually assessed blush grades for the prediction of follow-up LV-function in patients with CT99021 price acute ischemic syndromes. (c) 2010 Wiley-Liss, Inc.”
“Purpose: To determine the value of a topical carbonic anhydrase inhibitor

on the macular thickness and function in choroideremia patients with cystoid macular edema.\n\nMethods: https://www.selleckchem.com/products/p5091-p005091.html Two choroideremia patients with cystoid macular edema, observed by spectral-domain optical coherence tomography, were treated with a topical form of carbonic anhydrase inhibitor. Examinations performed before and during treatment included best-corrected visual acuity by using the Early Treatment Diabetic Retinopathy Study charts and contrast sensitivity measured with briefly presented grating targets and the Pelli-Robson letter contrast sensitivity chart, microperimetry, and spectral-domain optical coherence tomography.\n\nResults: The 2

choroideremia patients treated with dorzolamide 2% formulation had a noticeable reduction in macular thickness by spectral-domain optical coherence tomography. This reduction was found in both eyes after 2 months of treatment. After an additional 3 months of the same treatment regimen, a more noticeable reduction in macular thickness was observed. The two study patients had improvement of their visual acuity, in at least one eye, on Early Treatment Diabetic Retinopathy Study charts, but no clinically significant changes for the other measures of visual function.\n\nConclusion: The present study shows the potential efficacy of topical dorzolamide for treating choroideremia patients with cystoid macular edema.

Although adaptation to non-climatic heat sources (solar and geoth

Although adaptation to non-climatic heat sources (solar and geothermal) seems likely in some marine species, climate warming predictions

for marine ectotherms are largely based on adaptation to climatically relevant heat sources (air or surface sea water temperature). Here, we show that non-climatic solar heating underlies thermal resistance adaptation in a rocky-eulittoral-fringe snail. Comparisons of the maximum temperatures of the air, the snail’s body and the rock ARRY-470 substratum with solar irradiance and physiological performance show that the highest body temperature is primarily controlled by solar heating and re-radiation, and that the snail’s upper lethal temperature exceeds the highest climatically relevant regional air temperature by approximately 22 degrees C. Non-climatic thermal adaptation probably features widely among marine and terrestrial ectotherms and because it could enable species to tolerate climatic rises in air temperature, it deserves more consideration in general and for inclusion into climate warming models.”
“Case studies are based on actual medical negligence claims or medicolegal referrals; however

certain facts have been omitted or changed by the author to ensure the anonymity of the parties involved.\n\nThis article discusses a Medical Board complaint involving an allegation of failure to use an interpreter, AZD8055 datasheet resulting in the death of a patient, aged 35 years.”
“Purposes: The intercavernous sinuses (ICSs) connect both sides of the cavernous sinus. Knowledge of the ICSs is useful when employing the endoscopic endonasal transsphenoidal approach. Although there were some studies concerning the anatomy of ICSs using cadavers, there is no well-recognized method of the radiological evaluation

of the ICSs. Therefore, we evaluate the ICSs using computed tomography digital subtraction venography GDC-0994 price (CT-DSV) in the cases of unruptured aneurysm that had the normal sellae anatomy. Methods: We retrospectively analyzed 97 cases of unruptured cerebral aneurysm that had normal parasellar and sellar anatomy except for the aneurysms. The analysis was performed using the multi-planar reconstruction images obtained by CT-DSV in these cases. Results: The anterior ICS (AICS), inferior ICS (IICS) and posterior ICS (PICS) were detected in 51 cases (52.6%), 28 cases (28.9%) and 76 cases (78.4%) respectively. In the cases older than 51 years old, the AICS was detected significantly less often than in those younger than 50 years old. Conclusions: The CT-DSV images directly reflected the blood flow. Most of the past research on the ICSs was obtained using cadavers, which could therefore not difectly assess the normal human blood flow. The present study showed the usefulness of CT-DSV for evaluating the ICSs and to understand the normal anatomy of the ICSs. (C) 2015 Published by Elsevier B.V.


“Bmi1 is a polycomb group proto-oncogene

that has


“Bmi1 is a polycomb group proto-oncogene

that has been implicated in multiple tumor types. However, its role in hepatocellular carcinoma (HCC) development has not been well studied. In this article, we report that Bmi1 is overexpressed in human HCC samples. When Bmi1 expression is knocked down in human HCC cell lines, it significantly inhibits cell proliferation and perturbs cell cycle regulation. To investigate the role of Bmi1 in promoting liver cancer development in vivo, we stably expressed Bmi1 and/or an activated form of Ras (RasV12) in mouse liver. We found that while Bmi1 or RasV12 alone is not sufficient to promote liver cancer development, coexpression of Bmi1 and RasV12 promotes HCC formation in mice. Tumors induced by Bmi1/RasV12 resemble human HCC by deregulation of genes involved ALK targets in cell proliferation, apoptosis, and angiogenesis. Intriguingly, we found no evidence that Bmi1 regulates Ink4A/Arf expression in both in vitro and in vivo systems of liver tumor development. In summary, our study shows that Bmi1 can cooperate with other oncogenic signals to promote hepatic carcinogenesis in vivo. Yet Bmi1

functions independent of Ink4A/Arf repression in liver cancer development. (Mol Cancer Res 2009;7(12):1937-45)”
“The complete molecule of the title compound, C(24)H(30)O(6), is generated by a crystallographic inversion centre. In the unique part of the molecule, the four-atom -O-CH(2)-C(O)-O-chain between the benzene ring and the tert-butyl group

assumes a zigzag conformation BVD-523 [O-C-C-O torsion angle = -162.3 (1)degrees].”
“Growing BMS-345541 concentration evidence has demonstrated that microRNAs (miRNAs) play an important role in regulating cellular radiosensitivity. This study aimed to explore the role of miRNAs in non-Hodgkin’s lymphoma (NHL) radiosensitivity. Microarray was employed to compare the miRNA expression profiles in B cell lymphoma cell line Raji before and after a 2-Gy dose of radiation. A total of 20 differentially expressed miRNAs were identified including 10 up-regulated and 10 down-regulated (defined as P < 0.05). Among the differentially expressed miRNAs, miR-148b was up-regulated 1.53-fold in response to radiation treatment. A quantitative real-time polymerase chain reaction (PCR) assay confirmed the up-regulation of miR-148b after radiation. Transient transfection experiments showed that miR-148b was up-regulated by miR-148b mimic and down-regulated by miR-148b inhibitor in the Raji cells. A proliferation assay showed that miR-148b could inhibit the proliferation of Raji cells before and after radiation. A clonogenic assay demonstrated that miR-148b sensitized Raji cells to radiotherapy. MiR-148b did not affect the cell cycle profile of post-radiation Raji cells compared with controls.

Artificial limitations are set on the state transition probabilit

Artificial limitations are set on the state transition probabilities of the models to find efficient methods of parameter estimation. We apply these models to the simulation of the performance of Digital Video Broadcasting-Handheld (DVB-H). The parameters of the packet error models are approximated as functions of the time-variant received signal strength and speed of a mobile vehicular DVB-H receiver, and it is shown that useful results may be achieved with the described packet error models, particularly when simulating mobile reception in field conditions.”
“Resolution of acute

inflammation Ferroptosis inhibitor drugs is an active process locally controlled by a novel genus of specialized pro-resolving mediators (SPM) that orchestrate key resolution responses. Hence, it is of general interest to identify individual bioactive mediators and profile their biosynthetic pathways with related isomers as well as their relation(s) to classic cicosanoids in mammalian tissues. Lipid mediator (LM)-SPM levels and signature profiles of their biosynthetic pathways were investigated using liquid chromatography-tandem mass spectrometry (LC-MS-MS)-based LM metabololipidomics. LM and SPM were identified using bigger than = 6 diagnostic

ions and chromatographic behavior matching with both authentic and synthetic materials. This approach was validated using the composite reference plasma Selleckchem Stem Cell Compound Library (SRM1950) of 100 healthy individuals. Using targeted LM metabololipidomics, we profiled LM and SPM pathways in human peripheral blood

(plasma and serum) and lymphoid organs. In these, we identified endogenous SPM metabolomes, namely, the potent lipoxins (LX), resolvins (Rv), protectins (PD), and maresins (MaR). These included RvD1, RvD2, RvD3, MaR1, and NPD1/PD1, which were identified in amounts within their bioactive ranges. In plasma and serum, principal component analysis (PCA) identified signature profiles of eicosanoids and SPM clusters. Plasma-SPM increased with omega-3 and acetylsalicylic acid intake that correlated with increased phagocytosis of Escherichia coli in whole blood. These findings demonstrate an approach for identification of SPM pathways (e.g., resolvins, protectins, and maresins) selleck inhibitor in human blood and lymphoid tissues that were in amounts commensurate with their pro-resolving, organ protective, and tissue regeneration functions. LM metabololipidomics coupled with calibration tissues and physiological changes documented herein provide a tool for functional phenotypic profiling.”
“Objective. Given that the clinical features of several IgG4-related diseases (IgG4-RD) can mimic those of autoimmune disorders, the aim of this study was to find possible distinguishing characteristics that would help us identify such cases from the pool of patients in a rheumatology clinic. Methods. From our clinic’s medical records, we identified patients who fulfilled the recently published diagnostic criteria for IgG4-RD.

Three experienced radiologists visually assessed the image qualit

Three experienced radiologists visually assessed the image quality and they manually measured the stenotic severity.\n\nResults: Fewer measurement errors occurred with multisector reconstruction (p = 0.05), a slower HR (p < 0.001) and a larger luminal diameter (p = 0.014); measurement errors were not related with the observers or the stenotic severity. There was no significant difference in measurements as for the reconstruction

algorithms below an HR of 70 bpm. More nonassessable segments LY2090314 order were visualized with halfscan reconstruction (p = 0.004) and higher HRs (p < 0.001). Halfscan reconstruction had better quality scores when the HR was below 60 bpm, while multisector reconstruction had better quality scores when the HR was above 90 bpm. For the H PIS between 60 and 90 bpm, Bioactive Compound Library purchase both reconstruction modes had similar quality scores. With excluding the nonassessable segments, both reconstruction algorithms achieved a similar mean measured stenotic severity and similar

standard deviations.\n\nConclusion: At a higher HR (above 90 bpm), multisector reconstruction had better temporal resolution, fewer nonassessable segments, better quality scores better accuracy of measuring the stenotic severity in this phantom study.”
“Transpulmonary Atrial Pacing. Introduction: Patients with prior extracardiac (EC) conduit Fontan surgery are considered ineligible for transvenous atrial pacemaker implantation due to lack of venous access to the atrial myocardium. A new technique for atrial lead placement in the setting of this surgical anatomy is proposed. Methods and Results: A 30-year-old female with prior EC Fontan surgery, incessant

supraventricular ACY-738 nmr tachycardia, sinus node dysfunction, and multi-organ failure was admitted to our hospital. After placement of a transvenous lead in the left pulmonary artery (LPA) for temporary pacing with ensuing clinical improvement, the patient was taken back to the catheterization laboratory for definitive treatment. After ablation of the supraventricular tachycardia and 3D mapping of the common atrium, a puncture through the LPA and into the left-sided atrium was performed. A transvenous lead was then attached to the base of the left-sided appendage where excellent pacing and sensing characteristics were observed. Follow-up transesophageal echocardiography revealed stable lead placement and absence of pericardial effusion. The patient was discharged to home several days later on warfarin therapy. Conclusions: Transpulmonary access to the left-sided atrium for patients with prior EC Fontan surgery is a feasible strategy for transvenous atrial pacing. Lead placement in this location is associated with excellent pacing characteristics and involves a limited segment of lead within the pulmonary venous atrium. The approach is technically straightforward and avoids the need for surgical pacemaker placement.


“The lacrimal gland (LG) and superficial

gland of


“The lacrimal gland (LG) and superficial

gland of the third eyelid (SGTE) belong to accessory organs of the eye. The aim of the present studies was to evaluate the histological, histochemical and fine structure of the LG and SGTE obtained from 3 adult females and 2 adult males of alpaca (Vicugna pacos). The LG was situated in the dorsolateral angle of the orbit between the dorsal rectus and the lateral rectus muscles. The SGTE was located between the medial rectus muscle, the ventral rectus muscle and was partially covered by the ventral oblique muscle of the eyeball. There were no effect of gender on the morphometry of examined LG and SGTE. The third eyelid resembles an anchor in shape. During histological www.selleckchem.com/products/CAL-101.html www.selleckchem.com/products/3-deazaneplanocin-a-dznep.html and ultrastructural analyses using light and transmission electron microscopy, it was established that the LG and SGTE are tubulo-acinar glands with mucoserous characters. The LG contains either lymphocytes or plasma cells, while SGTE had rare plasma cells and numerous lymphocytes in connective

tissue. The cartilage of the third eyelid was composed of hyaline tissue. Numerous aggregations of lymphocytes as lymph nodules in bulbar surface of the third eyelid were observed. The LG and SGTE secretory cells exhibited a similar ultrastructure appearance in electron microscopic examination, with secretory cells tightly filled with intracytoplasmatic secretory granules and numerous clusters of mucus of different sizes which were observed in the peripheral cells compartment.”
“The main goal of this paper is to present the rovibrational energies and spectroscopic constants of the Cl-2 molecular system in the relativistic states , A’:(1)2 (u) , A:(1)1 (u) , and . More

precisely, we have evaluated the Cl-2 omega (e) , omega (e) x (e) , omega (e) y (e) , alpha (e) , gamma (e) and B (e) rovibrational spectroscopic constants using two different procedures. The first was obtained by combining the rovibrational energies, calculated through solving Schrodinger’s nuclear equation and the diatomic rovibrational energy equation. The second was obtained by using the Dunham method. The calculated properties are in good agreement with available experimental data.”
“Cytoskeleton assembly is instrumental PHA-739358 in the regulation of biological functions by physical forces. In a number of key cellular processes, actin filaments elongated by formins such as mDia are subject to mechanical tension, yet how mechanical forces modulate the assembly of actin filaments is an open question. Here, using the viscous drag of a microfluidic flow, we apply calibrated piconewton pulling forces to individual actin filaments that are being elongated at their barbed end by surface-anchored mDia1 proteins. We show that mDia1 is mechanosensitive and that the elongation rate of filaments is increased up to two-fold by the application of a pulling force.

Methods A user-friendly 10-item questionnaire was specifi cally

Methods. A user-friendly 10-item questionnaire was specifi cally developed. The scale

included the background date. From a predefi ned scale the patients were subgrouped into 3 categories in relation to SL: (1) “no SL” or “a few days of SL,” (2) “1 week of SL,” and (3)” more than 2 weeks of SL.” The Fisher exact test was used to compare categorical variables. Results. Twenty-three doctors examined 207 Caspase activity assay working patients. A total of 114 patients (56%) completed the follow-up questionnaire. The 10-item scale showed a good correlation between the total score at the fi rst general practitioner visit and predictable time of SL according to the 3 periods. The frequency of BR and referral to radiographical examination was low, and perhaps this was a consequence of using the scale. Conclusion. The specially developed short and user-friendly 10 item LBP scale was a good predictor of the duration of SL. A low rate of BR and radiographical examination may even be the result of using the scale.”
“Background: Preventing recurrence of depression forms an important challenge for current treatments. Cognitive control impairments often remain present during remission of depression, putting remitted depressed patients at heightened risk for new depressive episodes by disrupting emotion regulation

processes. Importantly, research indicates that cognitive control training targeting working memory functioning shows potential in reducing maladaptive emotion regulation and depressive symptomatology in clinically BX-795 depressed patients and at-risk student samples. The current study aims to test the effectiveness of cognitive control training as a preventive intervention in a remitted depressed sample, exploring effects of cognitive control training on rumination and depressive symptomatology, along with indicators of adaptive emotion regulation and functioning. Methods/design: We present a double

blind randomized controlled design. Remitted depressed adults will JNK-IN-8 complete 10 online sessions of a cognitive control training targeting working memory functioning or a low cognitive load training (active control condition) over a period of 14 days. Effects of training on primary outcome measures of rumination and depressive symptomatology will be assessed pre-post training and at three months follow-up, along with secondary outcome measure adaptive emotion regulation. Long-term effects of cognitive control training on broader indicators of functioning will be assessed at three months follow-up (secondary outcome measures). Discussion: This study will provide information about the effectiveness of cognitive control training for remitted depressed adults in reducing vulnerability for depression.

Further, chalcone 17 demonstrates

sub-micromolar to low m

Further, chalcone 17 demonstrates

sub-micromolar to low micromolar antiproliferative activity in LNCaP, MDA-PCa-2b, 22Rv1 and C4-2B prostate cancer cells, all of which express mutated ARs and confer resistance to the current clinically used antiandrogens. The results suggest that chalcone 17 could be a good candidate for further pre-clinical development as a novel antiandrogen for advanced prostate cancer.”
“Apoptosis-inducing S63845 in vivo factor (AIF) is a bifunctional mitochondrial flavoprotein critical for energy metabolism and induction of caspase-independent apoptosis, whose exact role in normal mitochondria remains unknown. Upon reduction with NADH, AIF undergoes dimerization and forms tight, long-lived FADH(2)-NAD charge-transfer complexes (CTC) that are proposed to be functionally important. To obtain a deeper insight into structure/function

relations and redox mechanism of this vitally important protein, we determined the X-ray structures of oxidized and NADH-reduced forms of naturally folded recombinant murine AIF. Our structures reveal that CTC with the pyridine nucleotide is stabilized by (i) pi-stacking interactions between coplanar nicotinamide, isoalloxazine, and Phe309 CP-456773 rings; (ii) rearrangement of multiple aromatic residues in the C-terminal domain, likely serving as an electron delocalization site; and (iii) an extensive hydrogen-bonding network involving His453, a key residue that undergoes a conformational switch to directly interact with and optimally orient the nicotinamide for charge transfer. Via the His453-containing peptide, redox changes in the active site are transmitted to the surface, promoting

AIF dimerization and restricting access to a primary nuclear localization selleck chemicals signal through which the apoptogenic form is transported to the nucleus. Structural findings agree with biochemical data and support the hypothesis that both normal and apoptogenic functions of AIF are controlled by NADH. (C) 2009 Elsevier Ltd. All rights reserved.”
“Purpose: Local failure in unresectable pancreatic cancer may contribute to death. We hypothesized that intensification of local therapy would improve local control and survival. The objectives were to determine the maximum tolerated radiation dose delivered by intensity modulated radiation with fixed-dose rate gemcitabine (FDR-G), freedom from local progression (FFLP), and overall survival (OS).\n\nMethods and Materials: Eligibility included pathologic confirmation of adenocarcinoma, radiographically unresectable, performance status of 0-2, absolute neutrophil count of >= 1500/mm(3), platelets >= 100,000/mm(3), creatinine <2 mg/dL, bilirubin <3 mg/dL, and alanine amino-transferase/aspartate aminotransferase <= 2.5 x upper limit of normal. FDR-G (1000 mg/m(2)/100 min intravenously) was given on days -22 and -15, 1, 8, 22, and 29. Intensity modulated radiation started on day 1. Dose levels were escalated from 50-60 Gy in 25 fractions.

These findings suggest that multitasking neurons provide a comput

These findings suggest that multitasking neurons provide a computational advantage for behaviors that place simultaneous demands on two or more cognitive processes.”
“Local residual disease occurs in 7-13 % after primary treatment for nasopharyngeal carcinoma (NPC). To prevent tumor progression and/or distant metastasis, treatment is indicated. Biopsy is the “gold standard” for diagnosing residual disease. Because late histological regression frequently is seen after primary treatment

for NPC, biopsy should be performed when imaging or endoscopy is suspicious at 10 weeks. Different modalities can be used in the treatment of local residual disease. Interestingly,

the treatment of residual disease has better outcomes than treatment of GDC 0032 recurrent disease. For early-stage disease (rT1-2), treatment results and survival rates are very good and comparable to patients who had a complete response after the first treatment. Surgery (endoscopic or open), brachytherapy (interstitial or intracavitary), external or 4-Hydroxytamoxifen cell line stereotactic beam radiotherapy, or photodynamic therapy all have very good and comparable response rates. Choice should depend on the extension of disease, feasibility of the treatment, and doctor’s and patient’s preferences and experience, as well as the risks of the adverse events. For the more extended tumors, choice of treatment is more difficult, because complete response rates are poorer and severe side effects are not uncommon. The results of external beam reirradiation and stereotactic radiotherapy are better than brachytherapy for T3-4 tumors. Photodynamic therapy resulted in good palliative responses in a few patients with extensive disease. Also, chemotherapeutics or the Epstein-Barr

virus targeted therapies can be used when curative intent treatment is not feasible anymore. However, their advantage in isolated local failure has not been well described yet. Because residual disease often is a problem in countries with Nutlin-3 solubility dmso a high incidence of NPC and limited radiotherapeutic and surgical facilities, it should be understood that most of the above mentioned therapeutic modalities (radiotherapy and surgery) will not be readily available. More research with controlled, randomized trials are needed to find realistic treatment options for residual disease.”
“The course of development of the human genital tract is undifferentiated to the 9th week of development. At this time two symmetrical paired ducts known as the mesonephric (MD) and paramesonephric ducts (PMD) are present, which together with the urogenital sinus provide the tissue sources for internal and external genital development.