The defence mechanisms of both individuals and insect colonies start with border defences to prevent parasite intake and are followed by soma defences that prevent the establishment and spread of the parasite between the body’s cells or the social insect workers. Lastly, germ line defences are employed to inhibit infection of the reproductive tissue of organisms or the reproductive individuals in colonies. We further find sophisticated self/non-self-recognition systems operating at both

levels, which appear to be vital in maintaining the integrity of the body or colony as a reproductive entity. We then expand on the regulation of immune responses and end with a contemplation of how evolution may shape the different immune components, both within and between levels. The aim of this review is to highlight common evolutionary principles acting in disease BI 6727 inhibitor defence at the level of both individual organisms and societies, thereby linking the fields of physiological and ecological immunology.”
“The Pacific whiteleg shrimp Litopenaeus

vannamei (Penaeidae) is one of the most important cultivated species in world aquaculture. In Brazil, the northeastern states are home to the main shrimp producers. As shrimp aquaculture has expanded and intensified, diseases have progressively become one of the most serious threats to this industry. Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is an enzootic viral agent in Brazilian shrimp farms. Its is usually diagnosed by histological methods. However, OSI-744 price to detect sub-clinical or acute IHHNV infection, more refined methods based on molecular techniques have been utilized. We found that by using “”universal”" primers and a single-step

PCR diagnostic test, it was difficult to distinguish between non-infective forms of the virus and active IHHNV. Detection of IHHNV was more accurate when we used two alternative molecular NU7441 cost strategies, namely 1) single-step PCR amplification based on gene choice and 2) reverse transcription coupled with PCR.”
“Background This study examined whether changes in self-efficacy explain the effects of a mailed print intervention on long-term dietary practices of breast and prostate cancer survivors. The relationship between change in self-efficacy and long-term physical activity (PA) also was examined. Methods Breast and prostate cancer survivors (N=543) from 39 US states and two Canadian provinces participated in the FRESH START intervention trial. Participants were randomly assigned to receive a 10-month program of mailed print materials on diet and PA available in the public domain or a 10-month program of tailored materials designed to increase fruit and vegetable (F&V) intake, decrease fat intake, and/or increase PA.

Different types of these phenomena have been ascribed to latency of perseveration, but direct evidence is lacking. We describe a patient with occipital lobe epilepsy who presented with both polyopia and palinopsia. We found that visual perseveration was related to the extent of movement of the original object and that the shape of the object changed in response to stimulation with another object. Palinopsia after the visual stimulus can be superimposed on the background picture, leading to polyopia. That polyopia and palinopsia are distinct phenomena is correct. but they seemed to be related. Both phenomena

occurred in the same patient and share an epileptic mechanism, In buy FG-4592 our patient, visual perseveration was part of an epileptic aura, developed when either the object or the patient moved, and diminished with gabapentin. (C) U0126 MAPK inhibitor 2009 Elsevier Inc. All rights reserved.”
“Objective-To determine the lowest ACTH dose that would induce a significant increase in serum cortisol concentration and identify the time to peak cortisol concentration in healthy neonatal foals.

Design-Prospective randomized

crossover study.

Animals-11 healthy neonatal foals.

Procedures-Saline (0.9% NaCl) solution or 1 of 4 doses (0.02, 0.1, 0.25, and 0.5 mu g/kg [0.009, 0.045, 0.114, and 0.227 mu g/lb]) of cosyntropin (synthetic ACTH) was administered IV. Serum cortisol concentrations were measured before and 10, 20, 30, 60, 90, 120, 180, and 240 minutes after administration of cosyntropin or saline solution; CBCs were performed before and 30, 60, 120, and 240 minutes after administration.

Results-Serum cortisol concentration was significantly increased, compared with baseline, by 10 minutes after cosyntropin administration at doses of 0.1, 0.25, and 0.5 mu g/kg. Serum cortisol concentration peaked 20 minutes after administration

of cosyntropin at doses of 0.02, 0.1, and 0.25 mu g/kg, with peak concentrations 1.7, 2.0, and 1.9 times the baseline concentration, respectively. Serum cortisol concentration peaked 30 minutes after cosyntropin administration at a dose of 0.5 mu g/kg, with peak concentration 2.2 times the baseline concentration. No significant differences were detected among peak serum cortisol concentrations obtained with Selleckchem MK-8776 cosyntropin administration at doses of 0.25 and 0.5 mu g/kg. Cosyntropin administration significantly affected the lymphocyte count and the neutrophil-to-lymphocyte ratio.

Conclusions and Clinical Relevance-Results suggested that in healthy neonatal foals, the lowest dose of cosyntropin to result in significant adrenal gland stimulation was 0.25 mu g/kg, with peak cortisol concentration 20 minutes after cosyntropin administration.”
“Study Design. An in vitro cadaveric study of interbody fusion devices (IFDs) with differing interface geometries. Objective.

(C) 2013 Elsevier Ireland Ltd. All rights reserved.”
“Spironolactone (SPR), a mineralocorticoid receptor blocker, diminishes hyperglycemia-induced reduction in glucose-6-phosphate dehydrogenase (G6PD) activity, improving oxidative stress damage. This study investigated whether SPR ameliorates nephropathy by increasing G6PD activity and reducing oxidative stress in spontaneously hypertensive diabetic rats (SHRs).

The DMH1 streptozotocin-induced diabetic rats received or not SPR 50 mg/kg per day, for eight weeks. A human mesangial cell line was cultured in normal or high glucose conditions, with or without SPR, for 24 h. Plasma glucose levels and systolic blood pressure were unaltered by diabetes or by SPR treatment. Albuminuria, fibronectin expression, 8-OHdG urinary levels, lipid peroxidation and p47phox expression were higher in the diabetic rats compared with the control and were reduced by SPR. The antioxidant GSH/GSSG ratio was reduced in the diabetic rats and the treatment reestablished it. Diabetes-induced

SGK1 up-regulation was inhibited by SPR. Reactive oxygen species (ROS) and superoxide production induced by NADPH oxidase were increased by hyperglycemia and high glucose, in vivo and in vitro, respectively, and were reduced with SPR. Hyperglycemia and high glucose decreased G6PD activity, which was restored with SPR. These results suggest that SPR ameliorates nephropathy in diabetic SHRs by restoring G6PD activity and diminishes oxidative stress without affecting glycaemia and blood pressure.”
“Narrow band CE-chirps (R) were

developed HSP990 to provide a better synchronization of neural response due to the compensation of the traveling wave delay in the basilar membrane. These stimuli combined with a detection method that includes higher response harmonics on the auditory steady-state response (ASSR) recording was studied in this research.

Objective: (1) To establish air conduction thresholds for ASSR to narrow band CE-chirp (R) in normal hearing full-term neonates; (2) describe the test time needed for the above in one ear and (3) to compare the results in infants and normally hearing adults.

Method: ASSR to air-conducted stimuli were obtained in 30 full-term neonates (14 girls and 16 boys) with an average selleck chemicals age of 343 h of life. All neonates were presented presence of transient-evoked otoacoustic emissions (TEOAE) and result “”pass”" in automatic ABR at 35 dB nHL before ASSR test. ASSR thresholds of both ears of 10 normal hearing adults (5 girls and 5 boys) varied in age between 23 and 30 years and with hearing thresholds better than or equal to 15 dB HL at all frequencies between 250 and 8000 Hz were recorded to compare with the neonate data.

Results: The neonate ASSR thresholds estimated from 50% using cumulative distributions were 24.5, 13.5, 7.5 and 10 dB nHL at 500, 1000, 2000, and 4000 Hz, respectively.

There was no improvement in lameness after diagnostic GDC-0994 in vivo local analgesia below

the carpal region, and 1 of 4 horses had mild improvement after cubital joint analgesia. Radiography revealed enthesophyte formation on the radial tuberosity and linear mineralization of the medial collateral ligament in 2 horses and periosteal reaction on the humeral condyle in all 4 horses. One horse had mild osteoarthritis of the cubital joint, and 3 had osteophytosis of the cranial aspect of the radius. Although all horses were initially examined because of an acute onset of lameness, all had chronic abnormalities visible on imaging. Ultrasonography revealed an irregular boney contour and enthesopathy at the insertion of the short medial collateral ligament to the radial tuberosity and desmitis of the short medial collateral ligament. Two horses had radiographic evidence of similar Staurosporine supplier but less severe lesions of the contralateral cubital joint.

Treatment and Outcome-All horses received phenylbutazone and rest. All horses were free of lameness after a median of 3 months (range, 2 to 4 months) and returned to previous use after a median of 6 months (range, 3 to 8 months).

Conclusions and Clinical Relevance-The results of the present report suggested, that performance horses with enthesopathy and desmitis of

the medial collateral ligament of the cubital joint may have a good prognosis for return to previous use following appropriate treatment. (J Am Vet Med Assoc 2013;242:1152-1158)”
“Background: To reduce the spread of drug-resistant pathogens, appropriate use of antimicrobials is all indispensable measure. Front January 2002, we undertook campaigns about antimicrobial treatment in Sado Island, Japan.

Methods: The Subjects were born in

1996 (group-1996) or during 2002-2004 group-2002-2004) and received outpatient treatment at our hospital. We evaluated the subjects’ medical information front patient records during their first 3 years of life. Sapitinib supplier Demographic data. duration of breast-feeding, and attending a day-care center (DCC) were evaluated using a questionnaire.

Results: Average visit-based antimicrobial prescription rates significantly decreased front 535 for group-1996, to 70 for group-2002-2004 per 1000 hospital visits (P < 0.001), The rate of cephalosporin prescriptions significantly decreased (77.1%-16.7%; P < 0.001), while amoxicillin (0.8%-29.5%) and macrolides (21.0%-52.4%) significantly increased (P < 0.001). Regarding 417 nasopharyngeal Cultures from group-2002-2004, 77.8% (179/ 230) of Streptococcus pneumoniae strains were nonsusceptible to penicillin. For Haemophilus influenzae strains, 30.6% (59/193) were nonsusceptible to ampicillin. Living with older sibling(s) and early DCC attendance were associated with carriage of resistant pathogens.

The first fFN test was selleckchem followed immediately by vaginal ultrasonography with a transvaginal probe ( three images per patient) and, thereafter, a second fFN test.

Results. The first fFN test was positive in three patients, and in all, the second, post-ultrasound, fFN test was also positive. In all women with a negative baseline fFN test (n = 25), the second, post-ultrasound, fFN test was also negative.

Conclusion. Vaginal ultrasound examination does not artificially change

the fFN status. This is in contrast to common understanding and may permit the performance of ultrasound examination before fFN, which can be restricted to cases of short cervix.”
“Background and Purpose: Physical inactivity is a leading public health concern, particularly among women and ethnic minority groups, where Latinas are among the largest and fastest growing U.S. populations. Acculturation, known to affect other health behaviors, may explain low physical activity (PA) among these underserved women. Research on the effects of acculturation on PA, however, is scarce or limited by methodology. The study purpose was to evaluate the association between

acculturation (i. e., language, birth country, and duration of U.S. residency) and PA in a national sample of women within the framework of the socioecologic model of health promotion.

Methods: A total of 5,861 women (86% white, mean age 37.2) were sampled

from Tubastatin A the Third National Health and Nutrition Examination Survey (NHANES III).

Results: Hierarchical multivariable regression modeling results indicate significant associations among language, duration of U.S. residency, and age, after controlling for confounders (all p < 0.05). Most women reported less than recommended PA.

Conclusions: These findings indicate that age, duration PND-1186 research buy of U.S. residency, and language are important to consider in combination when understanding women’s PA, findings that have implications for future research, theory, and clinical practice (e.g., making available PA assessments in multiple languages additionally tailored on age and investigating sociopolitical factors unique to Latinas).”
“Purpose: To evaluate the radial dilation force and basket opening dynamics of three small (<1.5F) stone baskets.

Materials and Methods: Boston Scientific OptiFlex (1.3F), Cook N-Circle Nitinol Tipless Stone Extractor (1.5F), and Sacred Heart Medical Halo (1.5F) baskets were tested for radial dilation force in 10 repetitions using a floating block atop a stationary block secured to a digital scale. The floating block, attached to a plastic frame and base, pressed down on the scale to measure the radial dilation of the open basket while being passed through the cylindrical opening between the blocks. These same baskets were tested in triplicate for basket width in 0.

Simple clinical signs were good predictors of outcome.”
“Purpose: To determine, by using contrast material-enhanced ultrasonography (US), how quickly renal tumors grafted in mice begin to revascularize after stopping bevacizumab

treatment.

Materials and Methods: All experiments were approved by the regional ethics committee. A human tumor cell line SK-NEP-1 was grafted at day 0 in the left kidney of 50 nude mice. Forty-two mice developed tumors and longitudinal follow-up was performed on 32 surviving mice. From day 13, 14 controls received biweekly saline; 11 mice received biweekly bevacizumab until day 35 (continuous); and seven received biweekly bevacizumab until day 22, then biweekly placebo until day 35 (discontinued). KPT-8602 clinical trial LBH589 ic50 Contrast-enhanced US was performed on days 13, 14, 22, 27, and 35. Once the injected contrast material

distribution reached an equilibrium phase, high acoustic pressure pulses were applied to destroy microbubbles in the capillary bed in the imaged plane. Reperfusion was monitored, and time-signal intensity (SI) curves were obtained from the linear average of SIs in intratumoral and matched-depth renal cortex regions of interest. A kinetic parameter calculated from reperfusion curves reflects local perfusion, normalized with respect to adjacent renal cortex perfusion. Normalized perfusion obtained from each group was compared with that from the other groups and with necrosis percentages and microvascular density assessed histologically at day 35. Comparisons were made by using analyses of variance and Tukey-Kramer tests.

Results: The lowest excised mean tumor weights (+/- standard deviation) corresponded to the longest bevacizumab-treatment duration: 1.4 g +/- 1.1 (continuous-treatment) compared with 2.3 g +/- 2.1 (discontinued) and 3.7 g +/- 1.9 (control) (P = .01). On day 35, the respective control and continuously treated groups had comparable and significantly larger necrotic areas: 37% +/- 14 and 32% +/- 17 larger than the discontinued-treatment group

(15% +/- 9; P < .05). Normalized perfusion increased significantly with time AZD1208 purchase (P = .02) in the discontinued-treatment group after therapy ceased (day 22).

Conclusion: Noninvasively measured contrast-enhanced US parameters demonstrated tumor revascularization after stopping antiangiogenic therapy in this murine tumor model.”
“Background: The main objective of this study was to determine the prevalence of multiple providers for different controlled substances using the largest electronic prescription monitoring program (PMP) in the United States. A secondary objective was to explore patient and medication variables associated with prescriptions involving multiple providers. PMPs monitor the final allocation of controlled substances from pharmacist to patient. The primary purpose of this scrutiny is to diminish the utilization of multiple providers for controlled substances.

Cell based experiments suggest a specific role for the IGF-1/IRS-2 signaling pathway in regulating alpha-/beta-secretase activity. Moreover circulating IGF-1 might influence A beta clearance from the brain by promoting A beta transport over the blood brain barrier. Interestingly, brain specific deletion of IRS-2 increases life span, suggesting that long term neuronal IGF-1R signaling might be harmful. Taken together, the data from humans and different model organisms indicate a role of IR/IGF-1R signaling

in A beta metabolism, and clearance as well as longevity. Since more studies are needed to elucidate the impact of insulin and/or IGF-1 treatment in AD, the time to propose these hormones as a potential treatment option for AD has not come yet.”
“Human milk from healthy women contains numerous nutrients such as antioxidants STA-9090 molecular weight which are necessary for newborns. AZD1480 The aim of this study was to evaluate the changes of total antioxidant capacity (TAC) and free

radical scavenging activity in human milk during the first six month period of lactation and also its relationship to maternal plasma. A total of 505 milk samples (colostrum, transitional and mature milks) collected from 115 healthy women with full term newborns. Blood plasma was obtained from 58 women at 3 months postpartum. The TAC of samples were measured by Ferric Reducing/Antioxidant Power assay and free radical scavenging activity were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. TAC was obviously higher in colostrums than transitional and mature milks. Similar results were ACY-241 observed for DPPH radical scavenging activity of the samples. There was a high significant correlation between the results of these two methods. The relationship

between the antioxidant content of human milk and maternal plasma was also significant. These data suggest that using colostrum, with high antioxidant potential during the first days of life is vital; moreover, reduction in total TAC during the course of lactation may needs more attention about nutritional status.”
“Alzheimer’s disease (AD) is a late-onset progressive neurodegenerative disorder which results in the irreversible loss of cortical neurons, particularly in the associative neocortex and hippocampus. AD is the most common form of dementia in the elderly. Apart from the neuronal loss, the pathological hallmarks are extracellular senile plaques, containing the peptide beta-amyloid (A beta) and neurofibrillary tangles. The A beta cascade hypothesis remains the main pathogenetic model, as suggested by familiar AD, mainly associated to mutation in amyloid precursor protein and presenilin genes. The remaining 95% of AD patients are mostly sporadic late-onset cases, with a complex aetiology due to interactions between environmental conditions and genetic features of the individual.

In the present study, frozen section diagnosis of adnexal masses in 282 patients was compared with permanent section diagnosis as the gold standard.

The overall accuracy of frozen section diagnosis was 95.7%. The sensitivity of frozen section

diagnosis for benign, borderline and malignant lesions was 99, 60, and 92%, respectively. The tumor size in discrepant cases was larger than the concordant cases, however no association between mucinous histology and inaccurate diagnosis was found. The sensitivities of gross examination and clinical data in distinguishing benign from non-benign lesions were 93 and 70%, respectively.

Frozen section is AZD8055 mw a reliable method for surgical management of patients with adnexal

masses. Gross examination and clinical data are two valuable parts of intraoperative consultation.”
“Purpose The inclusion of patient-reported outcome (PRO) instruments to record patient health-related quality of life (HRQOL) data has virtually become the norm in oncology randomised controlled trials (RCTs). Despite this fact, recent concerns have focused on the quality of reporting PKC412 molecular weight of HRQOL. The primary aim of this study was to evaluate the quality of reporting of HRQOL data from two common instruments in oncology RCTs.

A meta-review was undertaken of systematic reviews reporting HRQOL data collected using PRO instruments in oncology randomised controlled trials (RCTs). English language articles published between 2000 and 2012 were included and evaluated against a methodology checklist.

Four hundred SN-38 nmr and thirty-five potential articles were identified. Six systematic reviews were included in the analysis. A total of 70,403 patients had completed PROs. The European Organization for Research and Treatment of Cancer

QLQ-C30 and Functional Assessment of Cancer Therapy-General questionnaire accounted for 55 % of RCTs. Eighty per cent of RCTs had used psychometrically validated instruments; 70 % reported culturally valid instruments and almost all reported the assessment timing (96 %). Thirty per cent of RCTS reported clinical significance and missing data. In terms of methodological design, only 25 % of RCTs could be categorised as probably robust.

The majority of oncology RCTs has shortcomings in terms of reporting HRQOL data when assessed against regulatory and methodology guidelines. These limitations will need to be addressed if HRQOL data are to be used to successfully support clinical decision-making, treatment options and labelling claims in oncology.”
“Cognitive bizarreness is a shared feature of the dream and waking mentation of acutely psychotic patients. The authors investigated this measure of the structural architecture of thought in the dream and waking mentation of 20 nonpsychotic patients with Parkinson’s disease after treatment with prodopaminergic drugs.

The questionnaire consisted of 87 questions on broad aspects of general and psychiatric health. An additional questionnaire was enclosed for those respondents who reported experiencing UI. The data were analysed using binary logistic regression. The final study population constituted 4609 women, 1332 of whom had completed both questionnaires. The remaining 3277 had completed only the Life and Health questionnaire. Effect measures were odds ratios MLN4924 concentration (ORs) with corresponding 95% confidence intervals (CIs). Results. Statistically significant associations were found between UI and the occurrence of musculoskeletal pain (OR 1.45, 95% CI 1.20-1.76), fatigue

and sleeping disorders (OR 1.59, 95% CI 1.30-1.95), feelings of humiliation (OR 1.29, 95% CI 1.12-1.50), financial problems (OR 1.36, 95% CI 1.11-1.66), and reluctance to seek

medical care (OR 1.43, 95% CI 1.21-1.68). Conclusion. UI among women is commonly associated with a number of different psychosocial problems as well as an expressed feeling of vulnerability.”
“Aims and methods This study assessed incidence, predictive factors, Nutlin-3 nmr and outcome of Epstein-Barr virus (EBV) DNAemia in 100 recipients of allogeneic hematopoietic stem cell transplant. A total of 68 patients received anti-thymocyte globulin before unrelated grafts. Results Cumulative incidence of high-load EBV DNAemia defined by levels >10,000 copies/mL was 14% at 12months. In multivariate analysis, a CD4+ T-lymphocyte count >50L at day GSK1904529A +30 was the only factor significantly associated with a reduced risk of high-load EBV DNAemia. Thirteen of 16 patients with high viral loads were preemptively treated with rituximab and achieved EBV DNA negativity. Three patients had already developed

post-transplant lymphoproliferative disorder (PTLD) at the time of detection of high EBV DNA loads, and they obtained complete response after rituximab infusions and chemotherapy. Patients with high EBV DNA load had a significantly higher transplant-related mortality (TRM) compared with patients with negative or low viral load (54% vs. 16%, P=0.009) and a trend to lower overall survival (55% vs. 29%, P=0.060). Conclusion We conclude that CD4+ cell count at day +30 is a predictive factor for EBV DNAemia and may help identify patients requiring closer monitoring. Although only 3% of patients progressed to PTLD and were all successfully managed, EBV reactivation was associated with higher TRM, mainly because of infections.”
“Ten new 4-thiazolidinone derivatives have been synthesized from the alkanoic acids. Alkanoic acid hydrazides (I) on refluxing with aromatic aldehydes in the presence of a catalytic amount of glacial acetic acid in methanol furnish corresponding hydrazones (II). Thioglycolic acid react with (II) in methanol yield the title compounds (III). These 4-thiazolidinone derivatives were characterized by CHN analysis, IR and (1)H NMR spectral data.

27, 95% CI 0.11-0.64) correlated with lower mortality. There was a trend for lower mortality among patients started on antiplatelet drugs after admission (AOR 0.29, 95% CI 0.08-1.13). The effect of aspirin on mortality was much the same in patients who received 325 or 80 mg daily. Chronic antiplatelet therapy was not associated with a significantly lower risk of major embolism. In conclusion, chronic antiplatelet therapy was associated with lower mortality among patients with IE, independently of any effect

on major embolism. Whether or not a beneficial effect could be replicated by initiating antiplatelet therapy at the time of diagnosis remains unproven.”
“Background: Agua celeste, or “”heavenly water”", is the street name for a sky-blue colored solvent reportedly inhaled or ingested to produce an intoxicating effect. Study aims were to (1) describe prevalence of agua celestse (AC) use, and (2) identify correlates https://www.selleckchem.com/products/MK-2206.html of lifetime and recent use of AC use among female sex workers who also inject drugs (FSW-IDUs) in northern Mexico.

Methods: Between 2008 and 2010, baseline data from FSW-IDUs >= 18 years old living in Tijuana or Ciudad Juarez participating in a longitudinal behavioral intervention were analyzed using logistic regression.

Results: Among 623 FSW-IDUs (307 from Tijuana and 316 from Ciudad Juarez (CJ)), 166 (26%) reported ever using

AC, all of whom lived in CJ. Among the CJ sample, lifetime prevalence of AC use SBI-0206965 solubility dmso was 53%, median age of first use was 16 years (IQR: 14-23), and 10% reported it as their first abused substance. Ever using AC was independently associated with ever being physically abused and younger age, and was marginally associated with initiating injection drug use and regular sex work at age eighteen or younger. Among those ever using AC, 70/166(42.2%) reported using it within the last 6 months, which was independently associated

with using drugs with clients before or during sex, being on the street more than 8 h per day, and younger age.

Discussion: We observed considerable geographic variation in the use of AC in northern Mexico. Future studies exploring factors influencing use, its precise formulation(s), and its potential health effects are needed to guide prevention and treatment. (C) 2011 Elsevier Ireland Ltd. All rights reserved.”
“The present investigation deals www.selleckchem.com/products/VX-770.html with biological evaluation of Aster thomsonii. For this purpose, different biological assays of crude methanolic extract (CME) and its fractions that is n-Hexane fraction (NHF) and aqueous fraction (AQF) were carried out. The results of AQF showed maximum brine shrimp cytotoxic activity with ED(50) values of 154.69 mu g/ml, while the NHF showed significant potato disc antitumor activity with IC(50) values of 9.55 mu g/ml. Evaluation of both NHF and AQF fractions for sulforhodamine B assay on human cell line HT144 showed IC(50) values of 1.10 and 2.82 mg/ml, respectively.