There are no studies to support the clinical awareness of persistent Tanner stage (TS) 4 breast development in adulthood, and forensic experts continue to use TS 4 as evidence of age <18 years in cases of alleged child pornography.

One-fourth of nonclinical images of women over 18 years of age could be considered by a single forensic expert to represent TS 4. This observation, and substantial discordance in interpretation by pediatric endocrinologists, renders testimony based on this distinction invalid. (Read the full article)

Although the natural course of bilirubin levels has been extensively studied in general neonatal populations, there is a paucity of data regarding bilirubin trends in neonates before the development of significant hyperbilirubinemia.

This study provides data on the natural course of transcutaneous bilirubin before the development of significant hyperbilirubinemia, and on the effect of different demographic and perinatal risk factors on the rate of bilirubin increase in neonates with borderline bilirubin values. (Read the full article)

Social and environmental risks related to substandard housing contribute to adverse health outcomes. Partnerships between the health care and legal systems can help families address such risks and help clinicians understand the legal context of health.

A medical-legal partnership colocated in a pediatric primary care setting identified and treated a large cluster of poor quality, substandard housing. Housing improvements were possible because of strong collaboration between clinicians, attorneys, community partners, and families. (Read the full article)

Autism spectrum disorders are characterized by heterogeneous severity. Previous latent variable analyses of longitudinal data have focused on trajectories of related features such as IQ, and not on changes over time in standardized, observational measures of core autism symptoms.

Autism Diagnostic Observation Schedule–calibrated severity scores allow comparisons of observational data from toddlerhood to adolescence. This first report of latent autism severity trajectory classes indicates that most children show stability in core symptom severity over many years; small groups improved or worsened. (Read the full article)

National asthma treatment guidelines include the recommendation that all asthma patients receive a written asthma action plan. No previous study has sought to examine the readability, suitability, and content of asthma action plans within a nationally representative sample.

Although variability was found across written asthma action plans, and improvements in readability, suitability, and content are needed, there were also many common elements that would support a move to a single universal standard action plan. (Read the full article)

Birth asphyxia, or failure to initiate or sustain spontaneous breathing at birth, contributes to ~27% to 30% of neonatal deaths in resource-limited countries, including Tanzania. Without change, these countries will fail to meet Millennium Development Goal 4 targets by 2015.

The Helping Babies Breathe program was implemented in 8 hospitals in Tanzania in 2009. It has been associated with a sustained 47% reduction in early neonatal mortality within 24 hours and a 24% reduction in fresh stillbirths after 2 years. (Read the full article)

A policy of intubation, mechanical ventilation, and surfactant administration is commonly used for the treatment of respiratory distress syndrome worldwide; however subsequent development of bronchopulmonary dysplasia remains as risk with this standard approach.

Noninvasive surfactant administration technique during spontaneous breathing (Take Care) along with nasal continuous positive airway pressure support successfully reduces the need for further respiratory support and bronchopulmonary dysplasia rate in very low birth weight infants. (Read the full article)

UV radiation exposure in tanning beds is associated with an increased risk of skin cancer. Because of the rising rate of melanoma, the World Health Organization recommends that persons <18 years of age not use tanning devices.

Despite scientific evidence to the contrary, tanning facilities in Missouri, a state without indoor-tanning regulations, often misinformed consumers regarding the risk of skin cancer and would allow children as young as 10 years old to use tanning devices. (Read the full article)

Although the metabolic syndrome is associated with obesity, not all obese children have insulin resistance and metabolic syndrome, and nonobese children may develop these abnormalities. Associated factors have not been well described.

There was a 6.6% prevalence of nonobese children who were insulin-resistant, associated with a family history of hypertension. There was a 21.3% prevalence of obese who were not insulin-resistant, associated with a low waist circumference. (Read the full article)

There are no published prospective studies on the natural course and testicular growth in early childhood of spontaneously descended testes after birth compared with scrotal or surgically treated testes in boys with congenital cryptorchidism.

Data collected from this prospective study on the natural course and growth of the spontaneously descended testes add evidence-based data and recommendations on how to clinically manage boys with congenital cryptorchidism. (Read the full article)

Many US children aged 6 to 24 months who would be classified as low weight-for-age by the Centers for Disease Control and Prevention 2000 reference will be classified as normal weight-for-age by the World Health Organization 2006 standard.

Children who will be reclassified from low to normal weight-for-age using the World Health Organization growth standard are at higher risk of adverse health outcomes than children who are not low weight-for-age by the Centers for Disease Control and Prevention reference. (Read the full article)

Coxsackievirus A6 (CVA6) was identified as an important cause of "severe" hand, foot, and mouth disease (HFMD) during the 2011–2012 outbreak in North America. The atypical cutaneous features in this outbreak have not been well documented.

The cutaneous manifestations of CVA6-associated HFMD may be more extensive and variable than classic HFMD. Four distinct morphologies characterize this exanthem: (1) widespread vesiculobullous and erosive lesions, (2) "eczema coxsackium," (3) an eruption similar to Gianotti-Crosti, and (4) purpuric lesions. (Read the full article)

Evaluating for elevations in alanine aminotransferase (ALT) is a common screening test for the presence of nonalcoholic fatty liver disease (NAFLD). NAFLD is less common among non-Hispanic blacks. Better predictors of NAFLD are needed to identify individuals in most need of screening.

Relative to other ethnicities, metabolic syndrome and insulin resistance performed poorly at identifying non-Hispanic black adolescents with ALT elevations. The presence of metabolic syndrome may therefore not be an adequate trigger for NAFLD screening. Triglyceride elevations performed similarly between groups in identifying ALT elevations. (Read the full article)

Phthalate exposure has been associated with insulin resistance in animal studies and cross-sectional studies of adults, but has not been studied in adolescents.

We detect associations of urinary phthalate metabolites in a cross-sectional study of US adolescents. The association is highly robust to multiple sensitivity analyses, and specific to phthalates commonly found in food. Further longitudinal study of dietary phthalate exposures is needed. (Read the full article)

Invasive methicillin-resistant Staphylococcus aureus (MRSA) in children is associated with high morbidity and mortality. Although reductions in health care–associated MRSA infection among adults are documented, it is unclear if a similar trend is occurring among children.

Data from population-based surveillance were analyzed to assess changes in invasive MRSA infection incidence over time. This analysis describes the epidemiology and trends of invasive MRSA infections among children in 9 US metropolitan areas and estimates national burden. (Read the full article)

There is a perception that Gram-negative bacilli (GNB) bloodstream infection is increasing in the NICU, and those infections caused by a multidrug-resistant (MDR) strain are a growing threat to hospitalized patients.

Exposure to broad-spectrum antibiotics is the most important risk factor for MDR GNB bacteremia, which is associated with higher mortality. Neonates with risk factors for bacteremia caused by a MDR GNB strain may benefit from empirical antimicrobial therapy with carbapenem. (Read the full article)

Recent studies have shown that participating in the Supplemental Nutrition Assistance Program (SNAP) is associated with improved household food security. With the exception of 1 descriptive analysis, studies have not examined how SNAP affects children’s food security.

This article estimates the association between SNAP and children’s food security using the largest, most rigorous national study of food security to date. Given current proposals to reduce program size, this study underscores SNAP’s importance in affecting children’s well-being. (Read the full article)

Indoor tanning has gained widespread popularity among adolescents and young adults. Incidence rates of early-onset basal cell carcinoma also appear to be rising. Scant evidence exists on the impacts of early exposure and whether it leads to early occurrence of this malignancy.

In a US population-based study, indoor tanning was associated with an elevated risk of basal cell carcinomas occurring at or before the age of 50 years, with an increasing trend in risk with younger age at exposure among adolescents and young adults. (Read the full article)

Passive or opt-out consent has been successfully used to recruit subjects in several investigational studies. However, institutional review boards are often inconsistent in their application of federal regulations regarding passive consent.

This study documented the variability among 24 local institutional review boards in their application of federal regulations regarding passive consent and parental acceptance of a passive consent strategy in a multicenter pediatric study. (Read the full article)

Delaying cord clamping beyond 30 to 60 seconds after birth seems beneficial for all infants due to blood transfusion from placenta. Experimental data have demonstrated that ventilation implemented before cord clamping improved cardiovascular stability by increasing pulmonary blood flow.

Healthy self-breathing neonates in a low-resource setting are more likely to die if cord clamping occurs before or immediately after onset of spontaneous respirations. The risk of death/admission decreases by 20% for every 10-second delay in clamping after breathing. (Read the full article)

Phototherapy decreases bilirubin concentration in skin more rapidly than in blood. During and after phototherapy, transcutaneous bilirubin measurements are considered unreliable and therefore discouraged.

Transcutaneous bilirubin underestimates total serum bilirubin by 2.4 mg/dL (SD, 2.1 mg/dL) during the first 8 hours after phototherapy. This gives a safety margin of ~7 mg/dL below the treatment threshold to omit confirmatory blood sampling. (Read the full article)

Obesity among youth can have immediate health effects as well as longer-term consequences during adulthood. Overweight/obese children and adolescents are much more likely than normal-weight children to become overweight/obese adults.

This large, multisite longitudinal study examines patterns of exit from and entry into obesity between childhood and adolescence. Socioeconomic factors, body image, television habits, and parental obesity were important predictors of whether children remained obese or became obese. (Read the full article)

Antibiotic overuse is common and is a major public health threat. The prevalence of antimicrobial stewardship programs in children’s hospitals is growing. Single-center studies reveal that antimicrobial stewardship programs are effective in reducing unnecessary antibiotic use. Multicenter evaluations are needed.

Antibiotic use is declining overall across a large network of freestanding children’s hospitals. Hospitals with formalized antimicrobial stewardship programs experienced greater reductions in antibiotic use than other hospitals, suggesting that these interventions are an effective strategy to address antibiotic overuse. (Read the full article)

In most previous studies, transcutaneous bilirubin measurement has been found to provide an accurate estimate of total serum bilirubin levels. However, most of these studies were conducted in settings that optimized accuracy.

This study provides a "real-world" assessment of the accuracy of transcutaneous bilirubin measurements in multiple clinical settings and identification of sources of discrepancy between transcutaneous and total serum bilirubin measurements. (Read the full article)

Existing literature discusses the unique medical and psychological needs of sex trafficking victims and highlights the importance of screening patients with risk factors. However, little is known about providers’ knowledge and confidence in their ability to provide care to victims.

The study summarizes the knowledge gaps and barriers providers face when assisting pediatric sex trafficking victims. It also highlights the impact of training on providers’ confidence and ability to appropriately care for victims. (Read the full article)

Myxovirus resistance protein A (MxA) is a protein induced during viral infections. A few small-scale studies have suggested that MxA could be used as a marker of viral infection in clinical routine practice.

This study involves the largest patient population thus far and confirms the usefulness of MxA for diagnosing viral infections in children consulting the emergency department in a clinical routine setting. (Read the full article)

Insulin resistance plays a role in obesity. Recently it has been associated with increased risk of AD. Aβ42 and PSEN1 are molecules associated with increased risk of later AD. Patients affected by AD show elevated levels of plasma Aβ42.

Levels of Aβ42 and PSEN1 are significantly elevated in obese adolescents and correlated with the degree of both adiposity and systemic insulin resistance. (Read the full article)

Fifteen percent of tuberculosis cases globally are resistant to the drug isoniazid. Isoniazid resistance puts patients with tuberculosis at risk for poor treatment outcomes and threatens the effectiveness of isoniazid preventive therapy in people with latent tuberculosis infection.

We present the first global and regional estimates of the proportion of children with tuberculosis who have isoniazid-resistant disease, showing large geographic variations in risk of resistance. We estimate the number of annual incident cases of isoniazid-resistant tuberculosis in children. (Read the full article)

Parental hesitancy about childhood vaccines is prevalent and related to delay or refusal of immunizations. Physicians are highly influential in parental vaccine decision-making, but may lack confidence in addressing parents’ vaccine concerns.

A physician-targeted communications intervention designed to reduce maternal vaccine hesitancy through the parent-physician relationship did not affect maternal hesitancy or physician confidence communicating with parents. Further research should determine the most effective approaches to addressing vaccine hesitancy. (Read the full article)

Treatment of idiopathic steroid-resistant nephrotic syndrome is challenging, and therapeutic options are limited. In spite of good initial response with rituximab, responders always remain prone to further relapse, necessitating either repeat course of rituximab or addition of another steroid-sparing immunosuppressant.

Mycophenolate mofetil may be an effective maintenance therapy to consider as an additive immunosuppressant after induction with rituximab in maintaining remission among children with refractory steroid-resistant nephrotic syndrome. (Read the full article)

Research demonstrates that substance use, beliefs about use, and influence from family and friends can put young adults at risk for DUI and RWDD. Much of this research is cross-sectional and focuses on risk factors identified in young adults.

This longitudinal study emphasizes the importance of prevention efforts in early adolescence at multiple levels to reduce high school DUI/RWDD, because marijuana beliefs and marijuana use by peers and family members during middle school were associated with high school DUI/RWDD. (Read the full article)

A student-centered approach to teaching mathematics enables students to develop conceptual understanding and to grow as confident mathematicians.

The unexpected transition to online classes due to the COVID-19 pandemic has prompted many changes for undergraduate students and their instructors. To understand the magnitude of these impacts and potentially improve digital learning, researchers in the Penn State School of Engineering Design and Professional Programs have received a $196,136 grant from the National Science Foundation.

Source: www.businessinsider.com - Saturday, May 09, 2020
Dating apps like Tinder, Bumble, and Hinge have reported increased use amid the coronavirus pandemic, while touting virtual dating alternatives for users instead of meeting up in person. Swaths of users are still encouraging matches to break quarantine to have sex and go on dates, despite social distancing guidelines and fines to comply with them. An illustrator on Instagram has been collecting screenshots of these situations, and told Business Insider that users will brand themselves as "badasses," dispute the effectiveness of isolating, and lash out in anger and hurl abusive language when they're rejected. Spokespeople for Grindr, Tinder and Bumble told Business Insider they've informed users to adhere to social-distancing guidelines, but did not respond to inquiries about actions they're taking against users in places where violating lockdown orders can be against the law. Visit Business Insider's homepage for more stories . As millions remain confined to their homes to prevent the spread of coronavirus, the desire for human contact and connection has risen dramatically and led some to search for ways to break those social distancing rules. Popular dating apps — including Tinder , Bumble , and Hinge — have reported significant increases during the outbreak of swiping activity, matches between users, and messages exchanged. It's also led to the introduction of a breed of users who are interested in shirking lockdown orders, an

It took rickshaw driver Mohammad Rasheed a year to save 300 rupees to buy his daughter a bike, so when he found three billion rupees ($22.5 million) had passed through an unused bank account in his name, he was stunned ... and scared.

Trees, shrubs and woody vines are among the top food sources for honey bees in urban environments, according to an international team of researchers. By using honey bees housed in rooftop apiaries in Philadelphia, the researchers identified the plant species from which the honey bees collected most of their food, and tracked how these food resources changed from spring to fall. The findings may be useful to homeowners, beekeepers and urban land managers who wish to sustain honey bees and other bee and pollinator species.

La fièvre des élections s’empare de l’Afrique centrale. Pour la seconde fois depuis la fin des guerres qui ont dévasté la région, les scrutins s’enchaînent au Burundi, au Rwanda, en République centrafricaine et au Congo.

Election fever has spread across Central Africa. For the second time since the end of the disastrous civil wars in the region, electoral processes have been launched in Burundi, Rwanda, Central African Republic and the Congo.

Insufficient political will has thwarted regional efforts to stop the murderous Lord’s Resistance Army (LRA) but vigorous diplomacy led by the African Union (AU), an immediate military push and complementary civilian initiatives could end the misery of thousands.

The Framework Agreement signed by the UN, African organisations and eleven countries and the deployment of an intervention brigade in North Kivu are positive steps, but conflicts in the Kivu region of the Democratic Republic of the Congo also require a bottom-up approach aimed at improving intercommunal relations and restoring peace at the local level.

What should be included in a measure of school quality? If not by standardized test scores, how should we measure student achievement? Julian Vasquez Heilig and I each have some thoughts.

Background: This study summarizes drug resistance analyses in 4 recent phase 2b trials of the respiratory syncytial virus (RSV) fusion inhibitor presatovir in naturally infected adults.

Methods: Adult hematopoietic cell transplant (HCT) recipients, lung transplant recipients, or hospitalized patients with naturally acquired, laboratory-confirmed RSV infection were enrolled in 4 randomized, double-blind, placebo-controlled studies with study-specific presatovir dosing. Full-length RSV F sequences amplified from nasal swabs obtained at baseline and postbaseline were analyzed by population sequencing. Substitutions at RSV fusion inhibitor resistance-associated positions are reported.

Results: Genotypic analyses were performed on 233 presatovir-treated and 149 placebo-treated subjects. RSV F variant V127A was present in 8 subjects at baseline. Population sequencing detected treatment-emergent substitutions in 10/89 (11.2%) HCT recipients with upper and 6/29 (20.7%) with lower respiratory tract infection, 1/35 (2.9%) lung transplant recipients, and 1/80 (1.3%) hospitalized patients treated with presatovir; placebo-treated subjects had no emergent resistance-associated substitutions. Subjects with substitutions at resistance-associated positions had smaller decreases in viral load during treatment relative to those without, but similar clinical outcomes.

Conclusions: Subject population type and dosing regimen may have influenced RSV resistance development during presatovir treatment. Subjects with vs without genotypic resistance development had decreased virologic responses but comparable clinical outcomes.

QPX7728 is a new ultra-broad-spectrum inhibitor of serine and metallo beta-lactamases from a class of cyclic boronates that gave rise to vaborbactam. The spectrum and mechanism of beta-lactamase inhibition by QPX7728 were assessed using purified enzymes from all molecular classes. QPX7728 inhibits class A ESBLs (IC₅₀ range 1-3 nM) and carbapenemases such as KPC (IC₅₀ 2.9±0.4 nM) as well as class C P99 (IC₅₀ of 22±8 nM) with a potency that is comparable or higher than recently FDA approved BLIs avibactam, relebactam and vaborbactam. Unlike those other BLIs, QPX7728 is also a potent inhibitor of class D carbapenemases such as OXA-48 from Enterobacteriaceae and OXA enzymes from A. baumannii (OXA-23/24/58, IC₅₀ range 1-2 nM) as well as MBLs such as NDM-1 (IC₅₀ 55±25 nM), VIM-1 (IC₅₀ 14±4 nM) and IMP-1 (IC₅₀ 610±70 nM). Inhibition of serine enzymes by QPX7728 is associated with progressive inactivation with a high efficiency k₂/K ranging from of 6.3 x 10⁴ (for P99) to 9.9 x 10⁵ M^-1 s^-1 (for OXA-23). This inhibition is reversible with variable stability of the QPX7728-beta-lactamase complexes with target residence time ranging from minutes to several hours: 5-20 minutes for OXA carbapenemases from A. baumanii, ~50 minutes for OXA-48 and 2-3 hours for KPC and CTX-M-15. QPX7728 inhibited all tested serine enzymes at 1:1 molar ratio. Metallo-beta-lactamases NDM, VIM, and IMP were inhibited by a competitive mechanism with fast-on-fast-off kinetics, with K_is of 7.5±2.1 nM, 32±14 nM and 240±30 nM for VIM-1, NDM-1 and IMP-1, respectively. QPX7728 ultra-broad-spectrum of BLI inhibition combined with its high potency enables combinations with multiple different beta-lactam antibiotics.

Malaria parasites invade and replicate within red blood cells (RBCs), extensively modifying their structure and gaining access to the extracellular environment by placing the plasmodial surface anion channel (PSAC) into the RBC membrane. Expression of members of the cytoadherence linked antigen gene 3 (clag3) family is required for PSAC activity, a process that is regulated epigenetically. PSAC is a well-established route of uptake for large, hydrophilic antimalarial compounds and parasites can acquire resistance by silencing clag3 gene expression, thereby reducing drug uptake. We found that exposure to sub-IC50 concentrations of the histone methyltransferase inhibitor chaetocin caused substantial changes in both clag3 gene expression and RBC permeability, reversing acquired resistance to the antimalarial compound blasticidin S that is transported through PSAC. Chaetocin treatment also altered progression of parasites through their replicative cycle, presumably by changing their ability to modify chromatin appropriately to enable DNA replication. These results indicate that targeting histone modifiers could represent a novel tool for reversing epigenetically acquired drug resistance in P. falciparum.

Continuous spread of antimalarial drug resistance is a threat to current chemotherapy efficacy. Therefore, characterizing the genetic diversity of drug resistance markers is needed to follow treatment effectiveness and further update control strategies. Here, we genotyped Plasmodium falciparum resistance gene markers associated with sulfadoxine-pyrimethamine (SP) and artemisinin-based combination therapy (ACT) in isolates from pregnant women in Ghana. The prevalence of the septuple IRNI-A/FGKGS/T pfdhfr/pfdhps haplotypes including the pfdhps A581G and A613S/T mutations was high at delivery among post-SP treatment isolates (18.2%) compared to those of first-antenatal care (before initiation of intermittent preventive treatment of malaria in pregnancy with sulfadoxine-pyrimethamine (IPTp-SP); 6.1%; p = 0.03). Regarding the pfk13 marker gene, two non-synonymous mutations (N458D and A481C) were detected at positions previously related to artemisinin resistance in isolates from Southeast-Asia. These mutations were predicted in silico to alter the stability of the pfk13 propeller-encoding domain. Overall, these findings highlight the need for intensified monitoring and surveillance on additional mutations associated with increased SP resistance as well as emergence of resistance against artemesinin derivatives.

Since 2012, single low dose of primaquine (SLDPQ, 0.25mg/kg) has been recommended with artemisinin-based combination therapies, as first-line treatment of acute uncomplicated Plasmodium falciparum malaria, to interrupt its transmission, especially in low transmission settings of multidrug, including artemisinin, resistance. Policy makers in Cambodia have been reluctant to implement this recommendation due to primaquine safety concerns and lack of data on its efficacy.

In this randomized controlled trial, 109 Cambodians with acute uncomplicated P. falciparum malaria received dihydroartemisinin-piperaquine (DP) alone or combined with SLDPQ on the first treatment day. Transmission-blocking efficacy of SLDPQ was evaluated on Days 0, 1, 2, 3, 7, 14, 21, 28 and recrudescence by reverse transcriptase polymerase chain reaction (RT-PCR) (gametocyte prevalence) and membrane-feeding assays with Anopheles minimus mosquitoes (gametocyte infectivity). Without the influence of recrudescent infections, DP+SLDPQ reduced gametocyte carriage 3 fold compared to DP. Of 48 patients tested on Day 0, only three patients were infectious to mosquitoes (~6%). Post-treatment, three patients were infectious: on D14 (3.5%, 1/29), and on the first and seventh day of recrudescence (8.3%, 1/12 for each); this overall low infectivity precluded our ability to assess its transmission blocking efficacy.

Our study confirms effective gametocyte clearance of SLDPQ when combined with DP in multidrug resistant P. falciparum and the negative impact of recrudescent infections due to poor DP efficacy. Artesunate-mefloquine (ASMQ) has replaced DP and ASMQ-SLDPQ has been deployed to treat all P. falciparum symptomatic patients to further support the elimination of multidrug resistant P. falciparum in Cambodia.

Objectives: Antibiotic combination therapy is used for severe infections caused by multidrug-resistant (MDR) Gram-negative bacteria. Yet, data of which combinations are most effective is lacking. This study aimed to evaluate the in vitro efficacy of polymyxin B in combination with 13 other antibiotics against four clinical strains of MDR Pseudomonas aeruginosa.

Methods: We evaluated the interactions of polymyxin B in combination with amikacin, aztreonam, cefepime, chloramphenicol, ciprofloxacin, fosfomycin, meropenem, minocycline, rifampicin, temocillin, thiamphenicol or trimethoprim by automated time-lapse microscopy using predefined cut-off values indicating inhibition of growth (≤10⁶ CFU/mL) at 24 h. Promising combinations were subsequently evaluated in static time-kill experiments.

Results: All strains were intermediate or resistant to polymyxin B, anti-pseudomonal β-lactams, ciprofloxacin and amikacin. Genes encoding β-lactamases (e.g., bla_PAO and bla_OXA-50) and mutations associated with permeability and efflux were detected in all strains. In the time-lapse microscopy experiments, positive interactions were found with 39 of 52 antibiotic combination/bacterial strain setups. Enhanced activity was found against all four strains with polymyxin B used in combination with aztreonam, cefepime, fosfomycin, minocycline, thiamphenicol and trimethoprim. Time kill experiments showed additive or synergistic activity with 27 of the 39 tested polymyxin B combinations, most frequently with aztreonam, cefepime, and meropenem.

Conclusion: Positive interactions were frequently found with the tested combinations, also against strains that harboured several resistance mechanisms to the single drugs and with antibiotics that are normally not active against P. aeruginosa. Further study is needed to explore the clinical utility of these combinations.