Effect of nonnormal random components on level and power in group-randomized trials

The use of group-randomized trials is particularly widespread in the evaluation of health care, educational, and screening strategies. Group-randomized trials represent a subset of a larger class of designs often labeled nested, hierarchical, or multilevel and are characterized by the randomization of intact social units or groups, rather than individuals. The application of random effects models to group-randomized trials requires the specification of fixed and random components of the model. The underlying assumption is usually that these random components are normally distributed. This research is intended to determine if the Type I error rate and power are affected when the assumption of normality for the random component representing the group effect is violated. ^ In this study, simulated data are used to examine the Type I error rate, power, bias and mean squared error of the estimates of the fixed effect and the observed intraclass correlation coefficient (ICC) when the random component representing the group effect possess distributions with non-normal characteristics, such as heavy tails or severe skewness. The simulated data are generated with various characteristics (e.g. number of schools per condition, number of students per school, and several within school ICCs) observed in most small, school-based, group-randomized trials. The analysis is carried out using SAS PROC MIXED, Version 6.12, with random effects specified in a random statement and restricted maximum likelihood (REML) estimation specified. The results from the non-normally distributed data are compared to the results obtained from the analysis of data with similar design characteristics but normally distributed random effects. ^ The results suggest that the violation of the normality assumption for the group component by a skewed or heavy-tailed distribution does not appear to influence the estimation of the fixed effect, Type I error, and power. Negative biases were detected when estimating the sample ICC and dramatically increased in magnitude as the true ICC increased. These biases were not as pronounced when the true ICC was within the range observed in most group-randomized trials (i.e. 0.00 to 0.05). The normally distributed group effect also resulted in bias ICC estimates when the true ICC was greater than 0.05. However, this may be a result of higher correlation within the data. ^

Localization of Epileptogenic Zone on Pre-surgical Intracranial EEG Recordings: Toward a Validation of Quantitative Signal Analysis Approaches

In patients diagnosed with pharmaco-resistant epilepsy, cerebral areas responsible for seizure generation can be defined by performing implantation of intracranial electrodes. The identification of the epileptogenic zone (EZ) is based on visual inspection of the intracranial electroencephalogram (IEEG) performed by highly qualified neurophysiologists. New computer-based quantitative EEG analyses have been developed in collaboration with the signal analysis community to expedite EZ detection. The aim of the present report is to compare different signal analysis approaches developed in four different European laboratories working in close collaboration with four European Epilepsy Centers. Computer-based signal analysis methods were retrospectively applied to IEEG recordings performed in four patients undergoing pre-surgical exploration of pharmaco-resistant epilepsy. The four methods elaborated by the different teams to identify the EZ are based either on frequency analysis, on nonlinear signal analysis, on connectivity measures or on statistical parametric mapping of epileptogenicity indices. All methods converge on the identification of EZ in patients that present with fast activity at seizure onset. When traditional visual inspection was not successful in detecting EZ on IEEG, the different signal analysis methods produced highly discordant results. Quantitative analysis of IEEG recordings complement clinical evaluation by contributing to the study of epileptogenic networks during seizures. We demonstrate that the degree of sensitivity of different computer-based methods to detect the EZ in respect to visual EEG inspection depends on the specific seizure pattern.

Radiocarbon Analysis of Elemental and Organic Carbon in Switzerland during Winter-Smog Episodes from 2008 to 2012 – Part I: Source Apportionment and Spatial Variability

While several studies have investigated winter-time air pollution with a wide range of concentration levels, hardly any results are available for longer time periods covering several winter-smog episodes at various locations; e.g., often only a few weeks from a single winter are investigated. Here, we present source apportionment results of winter-smog episodes from 16 air pollution monitoring stations across Switzerland from five consecutive winters. Radiocarbon (14C) analyses of the elemental (EC) and organic (OC) carbon fractions, as well as levoglucosan, major water-soluble ionic species and gas-phase pollutant measurements were used to characterize the different sources of PM10. The most important contributions to PM10 during winter-smog episodes in Switzerland were on average the secondary inorganic constituents (sum of nitrate, sulfate and ammonium = 41 ± 15%) followed by organic matter (OM) (34 ± 13%) and EC (5 ± 2%). The non-fossil fractions of OC (fNF,OC) ranged on average from 69 to 85 and 80 to 95% for stations north and south of the Alps, respectively, showing that traffic contributes on average only up to ~ 30% to OC. The non-fossil fraction of EC (fNF,EC), entirely attributable to primary wood burning, was on average 42 ± 13 and 49 ± 15% for north and south of the Alps, respectively. While a high correlation was observed between fossil EC and nitrogen oxides, both primarily emitted by traffic, these species did not significantly correlate with fossil OC (OCF), which seems to suggest that a considerable amount of OCF is secondary, from fossil precursors. Elevated fNF,EC and fNF,OC values and the high correlation of the latter with other wood burning markers, including levoglucosan and water soluble potassium (K+) indicate that residential wood burning is the major source of carbonaceous aerosols during winter-smog episodes in Switzerland. The inspection of the non-fossil OC and EC levels and the relation with levoglucosan and water-soluble K+ shows different ratios for stations north and south of the Alps (most likely because of differences in burning technologies) for these two regions in Switzerland.

Visual dating of rockfall scars in Larix decidua trees

Dating past mass wasting with growth disturbances in trees is widely used in geochronology as the approach may yield dates of past process activity with up to subannual precision. Past work commonly focused on the extraction of increment cores, wedges, or stem cross sections. However, sampling has been shown to be constrained by sampling permissions, and the analysis of tree-ring samples requires considerable temporal efforts. To compensate for these shortcomings, we explore the potential of visual inspection of wound appearance for dating purposes. Based on a data set of 217 wood-penetrating wounds of known age inflicted to European larch (Larix decidua Mill.) by rockfall activity, we develop guidelines for the visual, noninvasive dating of wounds including (i) the counting of bark rings, (ii) a visual assessment of exposed wood and wound bark characteristics (such as the color and weathering status of wounds), and (iii) the relationship between wound age and tree diameter. A characterization of wounds based on photographs, randomly selected from the data set, reveals that young wounds typically can be dated with high precision, whereas dating errors gradually increase with increasing wound age. While visual dating does not reach the precision of dendrochronological dating, we clearly demonstrate that spatial patterns of and differences in rockfall activity can be reconstructed with both approaches. The introduction of visual dating approaches will facilitate fieldwork, especially in applied research, assist the conventional interpretation of tree-ring signals, and allow the reconstruction of geomorphic processes with considerably fewer temporal and financial efforts.

Comparison of the diagnostic significance of clinical, radiographic and ultrasonographic results after an experimental aerosol infection of pigs with Actinobacillus pleuropneumoniae

Scoring schemes for clinical, ultrasonographic and radiographic findings in pigs were developed based upon a standardized animal model for Actinobacillus pleuropneumoniae infection.The results of these methods were compared to each other as well as with the corresponding pathomorphological findings during necropsy. Altogether 69 pigs of different breeding lines (Hampshire, Pietrain and German Landrace were examined. Positive correlations were found between the results of all three methods as well as with the necropsy scores (p <0.0001). Different pathomorphological findings were detected either by radiographic or by ultrasonographic examination dependent upon the type of lung tissue alterations: Alterations of the pleura as well as sequestration of lung tissue on the lung surface could be clearly identified during the ultrasonographic examination while deep tissue alterations with no contact to the lung surface could be detected reliably by radiographic examination. Both methods complement each other, and the application of a combined ultrasonographic and radiographic examination of the thorax allows a comprehensive inspection of the lung condition. Particularly during the acute phase of the disease the extent of lung tissue damage can be estimated more precisely than by clinical examination alone.

An outbreak of Arthroderma vanbreuseghemii dermatophytosis at a veterinary school associated with an infected horse

We report a case of an outbreak of inflammatory dermatophytoses caused by Arthroderma vanbreuseghemii (formally Trichophyton mentagrophytes pro parte) that involved an infected horse, the owner and at least 20 students, staff and stablemen at a veterinary school in Bern (Switzerland) that presented highly inflammatory dermatitis of the body and the face. Transmission from human to human was also recorded as one patient was the partner of an infected person. Both the phenotypic characteristics and ITS sequence of the dermatophytes isolated from the horse and patients were identical, consistent with the conclusion that the fungus originated from the horse. Three infected persons had not been in direct contact with the horse. Although direct transmission from human to human cannot be ruled out, fomites were most likely the source of infection for these three patients. Inspection of the literature at the end of the nineteenth and beginning of the twentieth century revealed that this dermatophyte was frequently transmitted from horses to humans in contact with horses (stablemen, coachmen, carters and artillery soldiers). The rarity of the present case report at the present time is likely related to the transformation of civilisation from the nineteenth century to nowadays in Europe with the change of horse husbandry. In addition, the inadequate immune response of the horse and the high number of people in contact with it at the equine clinic may explain the exceptional aspect of this case report.

Inference of evolutionary forces acting on human biological pathways

Because natural selection is likely to act on multiple genes underlying a given phenotypic trait, we study here the potential effect of ongoing and past selection on the genetic diversity of human biological pathways. We first show that genes included in gene sets are generally under stronger selective constraints than other genes and that their evolutionary response is correlated. We then introduce a new procedure to detect selection at the pathway level based on a decomposition of the classical McDonald–Kreitman test extended to multiple genes. This new test, called 2DNS, detects outlier gene sets and takes into account past demographic effects and evolutionary constraints specific to gene sets. Selective forces acting on gene sets can be easily identified by a mere visual inspection of the position of the gene sets relative to their two-dimensional null distribution. We thus find several outlier gene sets that show signals of positive, balancing, or purifying selection but also others showing an ancient relaxation of selective constraints. The principle of the 2DNS test can also be applied to other genomic contrasts. For instance, the comparison of patterns of polymorphisms private to African and non-African populations reveals that most pathways show a higher proportion of nonsynonymous mutations in non-Africans than in Africans, potentially due to different demographic histories and selective pressures.

A simulation study on the statistical monitoring of condemnation rates from slaughterhouses for syndromic surveillance: an evaluation based on Swiss data

Syndromic surveillance (SyS) systems currently exploit various sources of health-related data, most of which are collected for purposes other than surveillance (e.g. economic). Several European SyS systems use data collected during meat inspection for syndromic surveillance of animal health, as some diseases may be more easily detected post-mortem than at their point of origin or during the ante-mortem inspection upon arrival at the slaughterhouse. In this paper we use simulation to evaluate the performance of a quasi-Poisson regression (also known as an improved Farrington) algorithm for the detection of disease outbreaks during post-mortem inspection of slaughtered animals. When parameterizing the algorithm based on the retrospective analyses of 6 years of historic data, the probability of detection was satisfactory for large (range 83-445 cases) outbreaks but poor for small (range 20-177 cases) outbreaks. Varying the amount of historical data used to fit the algorithm can help increasing the probability of detection for small outbreaks. However, while the use of a 0·975 quantile generated a low false-positive rate, in most cases, more than 50% of outbreak cases had already occurred at the time of detection. High variance observed in the whole carcass condemnations time-series, and lack of flexibility in terms of the temporal distribution of simulated outbreaks resulting from low reporting frequency (monthly), constitute major challenges for early detection of outbreaks in the livestock population based on meat inspection data. Reporting frequency should be increased in the future to improve timeliness of the SyS system while increased sensitivity may be achieved by integrating meat inspection data into a multivariate system simultaneously evaluating multiple sources of data on livestock health.

The Relationship of Student Demographics to 10th Grade MCAS Test Anxiety

The intensification of consequential testing situations is associated with an increase in anxiety among American students (Casbarro, 2005). Test anxiety can have negative effects on student test performance (Everson, Millsap, & Rodriguez, 1991). If test anxiety has the potential to decrease students’ test scores, it becomes a factor that can threaten the validity of any inferences drawn between test scores and student progress (Cizek & Burg, 2006). There are several factors that relate closely to test anxiety (Cizek & Burg, 2006). Variables of key influence include gender, socioeconomic status, and teacher-manifested anxiety (Hembree, 1988). Another influence upon test anxiety is students’ participation in academic support programs to prepare them for exit examinations. The purpose of this study was to examine the relationship between 10th grade high school student gender, socioeconomic status, perceived teacher anxiety, and student preparedness with levels of the Massachusetts Comprehensive Assessment System (MCAS) test anxiety. It appears that few studies have examined levels of high school test anxiety in regards to this specific high-stakes MCAS exit exam required for high school graduation. A two-phase sequential mixed-methods research design was used to survey (N=156) 10th grade students represented by a sampling of (n=80) students with low socioeconomic status and (n=76) students with high socioeconomic status regarding their levels of test anxiety in relation to upcoming MCAS testing. A multiple regression analysis was used to measure the relationship between the predictor variables (gender, socioeconomic status, perceived teacher anxiety, and student preparedness) with the criterion variable of student test anxiety using the Test Anxiety Inventory (TAI). Personal interviews with (n=20) volunteer students provided rich explanations of students’ academic self-efficacy, their perceptions of their performance on the upcoming MCAS exam, and their use of strategies to reduce their levels of test anxiety. Personal interviews with (n=12) volunteer school administrators and teachers provided descriptions of their perceptions of how test anxiety affected their students’ performance. A major quantitative finding of this study was that the variables of student socioeconomic status and student ratings of teacher anxiety accounted for the variance in students’ levels of surveyed test anxiety (R2 = .06, p = .033, small to medium effect size). These results indicate that different student populations vary in their readiness skills to successfully participate in consequential testing situations. Consequently, high-test anxious students would require emotional preparation as well as academic preparation when confronting high-stakes testing. The results have the potential to re-shape the format of schools’ MCAS test preparation efforts.

The Hospital Study of the City of Houston and Harris County

In 1941 the Texas Legislature appropriated $500,000 to the Board of Regents of the University of Texas to establish a cancer research hospital. The M. D. Anderson Foundation offered to match the appropriation with a grant of an equal sum and to provide a permanent site in Houston. In August, 1942 the Board of Regent of the University and the Trustees of the Foundation signed an agreement to embark on this project. This institution was to be the first one in the medical center, which was incorporated in October, 1945. The Board of Trustees of the Texas Medical Center commissioned a hospital survey to: - Define the needed hospital facilities in the area - Outline an integrated program to meet these needs - Define the facilities to be constructed - Prepare general recommendations for efficient progress The Hospital Study included information about population, hospitals, and other health care and education facilities in Houston and Harris County at that time. It included projected health care needs for future populations, education needs, and facility needs. It also included detailed information on needs for chronic illnesses, a school of public health, and nursing education. This study provides valuable information about the general population and the state of medicine in Houston and Harris County in the 1940s. It gives a unique perspective on the anticipated future as civic leaders looked forward in building the city and region. This document is critical to an understanding of the Texas Medical Center, Houston and medicine as they are today. SECTIONS INCLUDE: Abstract The Abstract was a summary of the 400 page document including general information about the survey area, community medical assets, and current and projected medical needs which the Texas Medical Center should meet. The 123 recommendations were both general (e.g., 12. “That in future planning, the present auxiliary department of the larger hospitals be considered inadequate to carry an added teaching research program of any sizable scope.”) and specific (e.g., 22. That 14.3% of the total acute bed requirement be allotted for obstetric care, reflecting a bed requirement of 522 by 1950, increasing to 1,173 by 1970.”) Section I: Survey Area This section basically addressed the first objective of the survey: “define the needed hospital facilities in the area.” Based on the admission statistics of hospitals, Harris County was included in the survey, with the recognition that growth from out-lying regional areas could occur. Population characteristics and vital statistics were included, with future trends discussed. Each of the hospitals in the area and government and private health organizations, such as the City-County Welfare Board, were documented. Statistics on the facilities use and capacity were given. Eighteen recommendations and observations on the survey area were given. Section II: Community Program This section basically addressed the second objective of the survey: “outline an integrated program to meet these needs.” The information from the Survey Area section formed the basis of the plans for development of the Texas Medical Center. In this section, specific needs, such as what medical specialties were needed, the location and general organization of a medical center, and the academic aspects were outlined. Seventy-four recommendations for these plans were provided. Section III: The Texas Medical Center The third and fourth objectives are addressed. The specific facilities were listed and recommendations were made. Section IV: Special Studies: Chronic Illness The five leading causes of death (heart disease, cancer, “apoplexy”, nephritis, and tuberculosis) were identified and statistics for morbidity and mortality provided. Diagnostic, prevention and care needs were discussed. Recommendations on facilities and other solutions were made. Section IV: Special Studies: School of Public Health An overview of the state of schools of public health in the US was provided. Information on the direction and need of this special school was also provided. Recommendations on development and organization of the proposed school were made. Section IV: Special Studies: Needs and Education Facilities for Nurses Nursing education was connected with hospitals, but the changes to academic nursing programs were discussed. The needs for well-trained nurses in an expanded medical environment were anticipated to result in significant increased demands of these professionals. An overview of the current situation in the survey area and recommendations were provided. Appendix A Maps, tables and charts provide background and statistical information for the previous sections. Appendix B Detailed census data for specific areas of the survey area in the report were included. Sketches of each of the fifteen hospitals and five other health institutions showed historical information, accreditations, staff, available facilities (beds, x-ray, etc.), academic capabilities and financial information.

Compilation of Thunnus thynnus (bluefin tuna) catches in the Northeast Atlantic Ocean 1950-2010

The data have been extracted and compiled from various sources but mainly from the ICES data base. The ICES data are from catch databases downloaded from the ICES website on 2014-01-14. These data are resolved by ICES area, country and year. During inspection of these data, it was noted that Norwegian data for years before 1950 had not been entered into the catch database on the ICES website. ICES has been notified of this omission by B. R. MacKenzie. The Norwegian data from ICES Bulletins. Statistiques has been added. Additional historical bluefin tuna catch data from other fishery reports and sources have been included in the data file for years preceding those when countries started reported their landings officially to ICES. These additional data have been reported in the literature previously (MacKenzie and Myers 2007, Fisheries Research).

Compilation of Thunnus thynnus (bluefin tuna) catche in the Northeast Atlantic Ocean 1906-1949

The data have been extracted and compiled from various sources but mainly from the ICES data base. The ICES data are from catch databases downloaded from the ICES website on 2014-01-14. These data are resolved by ICES area, country and year. During inspection of these data, it was noted that Norwegian data for years before 1950 had not been entered into the catch database on the ICES website. ICES has been notified of this omission by B. R. MacKenzie. The Norwegian data from ICES Bulletins. Statistiques has been added. Additional historical bluefin tuna catch data from other fishery reports and sources have been included in the data file for years preceding those when countries started reported their landings officially to ICES. These additional data have been reported in the literature previously (MacKenzie and Myers 2007, Fisheries Research).

Chronology and annual proxy data of Lago Grande di Monticchio sediments

We present new annual sedimentological proxies and sub-annual element scanner data from the Lago Grande di Monticchio (MON) sediment record for the sequence 76-112 thousand years before present (ka). They are combined with the previously published decadal to centennial resolved pollen assemblage in order to provide a comprehensive reconstruction of six major abrupt stadial spells (MON 1-6) in the central Mediterranean during early phase of the last glaciation. These climatic oscillations are defined by intervals of thicker varves and high Ti-counts and coincide with episodes of forest depletion interpreted as Mediterranean stadial conditions (cold winter/dry summer). Our chronology, labelled as MON-2014, has been updated for the study interval by tephrochronology and repeated and more precise varve counts and is independent from ice-core and speleothem chronologies. The high-resolution Monticchio data then have been compared in detail with the Greenland ice-core d18O record (NorthGRIP) and the northern Alps speleothem d18Ocalcite data (NALPS). Based on visual inspection of major changes in the proxy data, MON 2-6 are suggested to correlate with Greenland stadials (GS) 25-20. MON 1 (Woillard event), the first and shortest cooling spell in the Mediterranean after a long phase of stable interglacial conditions, has no counterpart in the Greenland ice core, but coincides with the lowest isotope values at the end of the gradual decrease in d18Oice in NorthGRIP during the second half of the Greenland interstadial (GI) 25. MON 3 is the least pronounced cold spell and shows gradual transitions, whereas its NorthGRIP counterpart GS 24 is characterized by sharp changes in the isotope records. MON 2 and MON 4 are the longest most and pronounced oscillations in the MON sediments in good agreement with their counterparts identified in the ice and spelethem records. The length of MON 4 (correlating with GS 22) supports the duration of stadial proposed by the NALPS timescales and suggests ca 500 yr longer duration than calculated by the ice-core chronologies GICC05modelext and AICC2012. Absolute dating of the cold spells provided by the MON-2014 chronology shows good agreement among the MON-2014, the GICC05modelext and the NALPS timescales for the period between 112 and 100 ka. In contrast, the MON-2014 varve chronology dates the oscillations MON 4 to MON 6 (92-76 ka) ca. 3,500 years older than the most likely corresponding stadials GS 22 to GS 20 by the other chronologies.

Seismic velocities and elastic properties of gabbroic rocks from ODP Hole 118-735B

The nearly continuous recovery of 0.5 km of generally fresh, layer 3 gabbroic rocks at Hole 735B, especially near the bottom of the section, presents scientists an unusual opportunity to study the detailed elastic properties of the lower oceanic crust. Extending compressional-wave and density shipboard measurements at room pressure, Vp and Vs were measured at pressures from 20 to 200 MPa using the pulse transmission method. All of the rocks exhibit significant increases in velocity with increasing pressure up to about 150 MPa, a feature attributed to the closing of microcrack porosity. Measured velocities reflect the mineralogical makeup and microstructures acquired during the tectonic history of Hole 735B. Most of the undeformed and unaltered gabbros are approximately 65:35 plagioclase/clinopyroxene rocks plus olivine or oxide minerals, and the observed densities and velocities are fully consistent with the Voigt-Reuss-Hill (VRH) averages of the component minerals and their proportions. Depending on their olivine content, the predominant olivine gabbros at 200 MPa have average Vp = 7.1 ± 0.2 km/s, Vs = 3.9 ± 0.1 km/s, and grain densities of 2.95 ± 0.5 g/cm3. The less abundant iron-titanium (Fe-Ti) oxide gabbros average Vp = 6.75 ± 0.15 km/s, Vs = 3.70 ± 0.1 km/s, and grain densities of 3.22 ± 0.05 g/cm3, reflecting the higher densities and lower velocities of oxide minerals compared to olivine. About 30% of the core is plastically deformed, and the densities and directionally averaged velocities of these shear-zone tectonites are generally consistent with those of the gabbros, their protoliths. Three sets of observations indicate that the shear-zone metagabbros are elastically anisotropic: (1) directional variations in Vp, both vertical and horizontal and with respect to foliation and lineation; (2) discrepancies among Vp values for the horizontal cores and the VRH averages of the component minerals and their mineral proportions, suggesting preferred crystallographic orientations of anisotropic minerals; and (3) variations of Vs of up to 7%, with polarization directions parallel and perpendicular to foliation. Optical inspection of thin sections of the same samples indicates that plagioclase feldspar, clinopyroxene, and amphibole typically display crystallographic-preferred orientations, and this, plus the elastic anisotropy of these minerals, suggests that preferred orientations are responsible for much of the observed anisotropy, particularly at high pressure. Alteration tends to be localized to brittle faults and brecciated zones, and typical alteration minerals are amphibole and secondary plagioclase, which do not significantly change the velocity-density relationships.

Occurence of foraminifera and other microfossils in sediment core CRP-2/2A (Table 1)

Sparse to moderately abundant foraminiferal assemblages from Oligocene and Lower Miocene sediments in the CRP-2/2A drillhole contain C.27 genera and 42 species of calcareous benthic foraminifera. No planktic or agglutinated taxa were observed. On the basis of their faunal characteristics, four Foraminiferal Units are defined in drillhole succession: Foraminiferal Unit I (26.91-193.95 mbsf), mostly sparse assemblages with Elphidium magellanicum and Cribroelphidium sp.; Foraminiferal Unit II (193.95-342.42 mbsf), mostly moderately abundant assemblages with Cassidulinoides aequilatera and Eponides bradyi; Foraminiferal Unit III (342.42-486.19 mbsf), moderately abundant to sparse assemblages characterised by Cassidulinoides chapmani and Stainforthia sp.; and Foraminiferal Unit IV, Improverished (486.19-624.15, total depth, mbsf), with mostly barren residues, but with large Milioliidae recorded in situ at various horizons in the drill core. Foraminiferal Units I-IV lack taxa allowing correlation to standard zonal schemes. Inspection of faunal records from CIROS-1 and DSDP 270 indicates that, although the faunas show an overall similarity, CRP-2/2A Foraminiferal Units I-IV are not identifiable at these sites. The units are therefore most likely to reflect local environmental changes, and probably will prove useful for local correlation, but their lateral extent is undetermined. All four assemblages apparently represent various glacially-influenced shelf environments, and appear to reflect a long term deepening trend from Units IV to II, from perhaps inner to mid or outer-shelf depths, followed by a return to shallower, inner shelf, conditios for Unit I.