On the other hand, downregulation of PTPN11 expression blocked the transcriptional activation of genes (p21, p15, p16, RB1, and IGFBP7) involved in cell cycle regulation and oncogene-induced senescence in response to BRAF V600E expression. Therefore, it can be assumed that SHP-2 participates not only in emergence of drug resistance in cancer cells, but also in oncogene-induced cell senescence.Poly(ADP-ribose) polymerase 1 (PARP-1) is a key DNA repair enzyme and an important target in cancer treatment. Conventional methods of studying the reaction mechanism of PARP-1 have limitations because of the complex structure of PARP-1 substrates; however, the necessary data can be obtained by molecular modeling. In this work, a molecular dynamics model for the PARP-1 enzyme-substrate complex containing NAD+ molecule and the end of the poly(ADP-ribose) chain in the form of ADP molecule was obtained for the first time. Interactions with the active site residues have been characterized where Gly863, Lys903, Glu988 play a crucial role, and the SN1-like mechanism for the enzymatic ADP-ribosylation reaction has been proposed. Models of PARP-1 complexes with more sophisticated two-unit fragments of the growing polymer chain as well as competitive inhibitors 3-aminobenzamide and 7-methylguanine have been obtained by molecular docking.In our previous papers, we proposed the idea that programs predicting intrinsically disordered regions in amino acid sequences can be used for finding weakened sites in proteins. The regions predicted by such programs are suitable targets for the introduction of protein-stabilizing mutations. However, for each specific protein, it remains unclear what determines protein stabilization - the amino acid sequence (and accordingly, prediction of weakened sites) or the 3D structure. To answer this question, it is necessary to study two proteins with similar structures but different amino acid sequences and, consequently, different predictions of weakened regions. https://www.selleckchem.com/products/ly2801653-merestinib.html By introducing identical mutations into identical elements of the two proteins, we will be able to reveal whether predictions of the weakened sites or the 3D protein structure are the key factors in the protein stability increase. Here, we have chosen ribosomal proteins L1 from the halophilic archaeon Haloarcula marismortui (HmaL1) and extremophilic bacterium Aquifex aeolicus (AaeL1). These proteins are identical in their structure but different in amino acid sequences. A disulfide bond introduced into the region predicted as the structured one in AaeL1 did not lead to the increase in the protein melting temperature. At the same time, a disulfide bond introduced into the same region in HmaL1 that was predicted as a weakened one, resulted in the increase in the protein melting temperature by approximately 10°C.Here, we determined qualitative and quantitative characteristics of the chaperone and immunoglobulin-binding activities of recombinant Skp protein (rSkp) from Yersinia pseudotuberculosis using the methods of dynamic light scattering and surface plasmon resonance. Commercial human polyclonal IgG and Fc and Fab fragments of human IgG were used as substrate proteins. The activity of rSkp strongly depended on the medium pH. The most stable low-molecular-weight complexes with a hydrodynamic radius up to 10 nm were formed by rSkp and protein substrates at acidic pH values. Under these conditions, rSkp exhibited the lowest propensity to self-association and the highest affinity for human IgG and its Fc and Fab fragments, as well as prevented their aggregation most efficiently (i.e., demonstrated the maximal chaperone activity). As the medium pH increased, the affinity of rSkp for IgG and its fragments decreased; rSkp was not able to completely prevent the aggregation of protein substrates, but significantly slowed it down. The obtained information may be of practical interest, since the stability of therapeutic IgG preparations affects their safety and efficacy in medical applications.Human genome contains ca. 20,000 protein-coding genes that could be translated into millions of unique protein species (proteoforms). Proteoforms coded by a single gene often have different functions, which implies different protein partners. By interacting with each other, proteoforms create a network reflecting the dynamics of cellular processes in an organism. Perturbations of protein-protein interactions change the network topology, which often triggers pathological processes. Studying proteoforms is a relatively new research area in proteomics, and this is why there are comparatively few experimental studies on the interaction of proteoforms. Bioinformatics tools can facilitate such studies by providing valuable complementary information to the experimental data and, in particular, expanding the possibilities of the studies of proteoform interactions.KLF2 is a member of the Krüppel-like transcription factor family of proteins containing highly conserved DNA-binding zinc finger domains. KLF2 participates in the differentiation and regulation of the functional activity of monocytes, T lymphocytes, adipocytes, and vascular endothelial cells. The activity of KLF2 is controlled by several regulatory systems, including the MEKK2,3/MEK5/ERK5/MEF2 MAP kinase cascade, Rho family G-proteins, histone acetyltransferases CBP and p300, and histone deacetylases HDAC4 and HDAC5. Activation of KLF2 in endothelial cells induces eNOS expression and provides vasodilatory effect. Many KLF2-dependent genes participate in the suppression of blood coagulation and aggregation of T cells and macrophages with the vascular endothelium, thereby preventing atherosclerosis progression. KLF2 can have a dual effect on the gene transcription. Thus, it induces expression of multiple genes, but suppresses transcription of NF-κB-dependent genes. Transcription factors KLF2 and NF-κB are reciprocal antagonists. KLF2 inhibits induction of NF-κB-dependent genes, whereas NF-κB downregulates KLF2 expression. KLF2-mediated inhibition of NF-κB signaling leads to the suppression of cell response to the pro-inflammatory cytokines IL-1β and TNFα and results in the attenuation of inflammatory processes.

0125), and significant relationships for hearing aid benefit were absent with only few exceptions. Hearing aid benefit defined by global APHAB using NU-6 and SPIN scores showed significant but weak positive correlations (r = 0.37, p less then 0.001; r = 0.28, p = 0.005, respectively) and ease of communication APHAB subscale scores (r = 0.32, p less then 0.001; r = 0.33, p = 0.001, respectively). CONCLUSION Hearing aid benefit assessed with audiologic measures were poor predictors of patient-reported benefit. Thus, patient-reported outcomes may provide a unique assessment of patient-perceived benefit from hearing aids, which can be used to direct hearing aid programming, training, or recommendations of alternative hearing services. © 2020 S. Karger AG, Basel.In this study, a transparent particle-free reactive silver ink was used to fabricate conductive patterns on flexible substrate. Thermal annealing and plasma irradiation at low temperature were utilized to improve the conductivity of as-printed patterns. The effects of sintering process parameters on the microstructure and resistivity of patterns were investigated. Under the optimized process conditions, the resistivity of pattern reached 1.2 ×10 -7 Ω · m by thermal sintering, while it was 8 ×10 -8 Ω · m after plasma sintering. Combined with these two sintering techniques, the resistivity was reduced to 6 × 10 -8 Ω · m, close to that of bulk silver. This work provides an alternative solution for fabricating of highly conductive feature patterns on common flexible substrates. © 2020 IOP Publishing Ltd.The poor conductivity of sulfur and the "shuttle effect" of polysulfide intermediates have hindered the development of next generation lithium-sulfur (Li-S) batteries with high energy and low consumption. Herein, novel Co9S8-S composite nanotubes are developed to efficiently alleviate above-mentioned problems. Experiments and theoretical calculations show that Co9S8 has strong adsorption on soluble polysulfides. This can not only restrain polysulfides diffusion and ensure their utilization, but also enhance the intimate contact between the active materials and the conductive substrates to promote the kinetics of conversion reaction. The three-dimensional (3D) conductive network with high surface area formed by interlinking Co9S8 nanotubes further improves the electronic conductivity of the composite cathode. https://www.selleckchem.com/products/CP-690550.html As a result, the Co9S8-S cathode shows a high capacity of 1153 mAh g-1. After 500 cycles, it shows a high capacity of 462 mAh g-1 (2 C) with a negligible capacity decay of ~0.04% per cycle. © 2020 IOP Publishing Ltd.Hierarchical actuators are comprised of multiple individual actuator elements arranged into a system, resulting in improved and expanded performance. Natural muscle tissue is a complex and multi-level example of hierarchical actuation, with its hierarchy spanning from the micrometer to the centimeter scale. In addition to a hierarchical configuration, muscle tissue exists in varying geometric arrangements. Pennate muscle tissue, denoted by its characteristic fibers extending obliquely away from the muscle tissue line of action, leverages geometric complexity to transform the relationship between fiber inputs and muscle tissue outputs. In this paper, a bioinspired hierarchical pennate actuator is detailed. This work expands on previous pennate actuator studies by deriving constitutive force, contraction, and stiffness models for a general pennate actuator, where the constituent fibers can be constructed from any linear actuator. These models are experimentally validated by studying a pennate actuator with McKibben artificial muscles constituting the actuator fibers. McKibben artificial muscles are used because they have a high force-to-weight ratio and are inexpensive to construct, making them an attractive candidate for hierarchical actuators and mobile robotics. Using the derived constitutive models, general pennate actuator performance is better understood by analyzing the transmission ratio, blocked force, and free contraction. Loaded contractions and stiffness during isotonic and isobaric contractions are also explored. The results allow for informed design decisions and an understanding of the associated tradeoffs when recreating the remarkable properties of pennate musculature. Future work will leverage the results of this paper to create an adaptive pennate actuator that is capable of changing configuration in response to force, contraction, and stiffness demands. © 2020 IOP Publishing Ltd.Controlling sub-microsecond desorption of water and other impurities from electrode surfaces at high heating rates is crucial for pulsed power applications. Despite the short time scales involved, quasi-equilibrium ideas based on transition state theory (TST) and Arrhenius temperature dependence have been widely applied to fit desorption barriers. In this work, we apply molecular dynamics (MD) simulations in conjunction with equilibrium potential-of-mean-force (PMF) techniques to directly compute the free energy barrier (Delta G*) associated with desorption of intact water molecules from Fe(2)O(3) and Cr(2)O(3) (0001) surfaces. The desorption free energy profiles are diffuse, without maxima, and have substantial dependences on temperature and surface water coverage. Incorporating the predicted Delta G* into an analytical form gives rate equations that are in reasonable agreement with non-equilibrium molecular dynamics desorption simulations. We also show that different Delta G* analytical functional forms which give similar predictions at a particular heating rate can yield desorption times that differ by up to a factor of four or more when the ramp rate is extrapolated by 8 orders of magnitude. This highlights the importance of constructing a physically-motivated Delta G* functional form to predict fast desorption kinetics. © 2020 IOP Publishing Ltd.Single-layer FeSe films grown on SrTiO3, with the highest superconducting transition temperature (TC) among all the iron-based superconductors, serves as an ideal platform for studying the microscopic mechanisms of high-TCsuperconductivity. The significant role of interfacial coupling has been widely recognized, while the precise nature of theTCenhancement remains open. In this review, we focus on the investigations of the interfacial coupling in FeSe/SrTiO3from the perspective of electron-phonon coupling (EPC). The main content will include an overview of the experimental measurements associated with different theoretical models and arguments about the EPC. Especially, besides the discussions of EPC based on the measurements of electronic states, we will emphasize the analyses based on phonon measurements. A uniform picture about the nature of the EPC and its relation to theTCenhancement in FeSe/SrTiO3has still not achieved, which should be the key for further studies aiming to the in-depth understanding of high-TCsuperconductivity and the discovery of new superconductors with even enhancedTC.

Bioinformatics analysis showed that both PAI-1 and uPAR were correlated with EP3 expression, as well as the prognosis of cervical cancer patients. The survival analysis further showed that uPAR overexpression (IRS≥2) was correlated with a lower overall survival rate of cervical cancer patients with advanced stages (FIGO III-IV). Conclusion These results indicated that EP3 signaling pathway might facilitate the migration of cervical cancer cells through modulating uPAR expression. Therefore, EP3 and uPAR could represent novel therapeutic targets in the treatment of cervical cancer in advantaged stages.Background Rice blast is an economically important and mutable disease of rice. Using host resistance gene to breed resistant varieties has been proven to be the most effective and economical method to control rice blast and new resistance genes or quantitative trait loci (QTLs) are then needed. Results In this study, we constructed two advanced backcross population to mapping blast resistance QTLs. CR071 and QingGuAi3 were as the donor parent to establish two BC3F1 and derived BC3F2 backcross population in the Jin23B background. By challenging the two populations with natural infection in 2011 and 2012, 16 and 13 blast resistance QTLs were identified in Jin23B/CR071 and Jin23B/QingGuAi3 population, respectively. Among Jin23B/CR071 population, 3 major and 13 minor QTLs have explained the phenotypic variation from 3.50% to 34.08% in 2 years. And, among Jin23B/QingGuAi3 population, 2 major and 11 minor QTLs have explained the phenotypic variation from 2.42% to 28.95% in 2 years. Conclusions Sixteen and thirteen blast resistance QTLs were identified in Jin23B/CR071 and Jin23B/QingGuAi3 population, respectively. QTL effect analyses suggested that major and minor QTLs interaction is the genetic basis for durable blast resistance in rice variety CR071 and QingGuAi3.Endophytic microbes are known to live asymptomatically inside their host throughout different stages of their life cycle and play crucial roles in the growth, development, fitness, and diversification of plants. The plant-endophyte association ranges from mutualism to pathogenicity. These microbes help the host to combat a diverse array of biotic and abiotic stressful conditions. Endophytic microbes play a major role in the growth promotion of their host by solubilizing of macronutrients such as phosphorous, potassium, and zinc; fixing of atmospheric nitrogen, synthesizing of phytohormones, siderophores, hydrogen cyanide, ammonia, and act as a biocontrol agent against wide array of phytopathogens. Endophytic microbes are beneficial to plants by directly promoting their growth or indirectly by inhibiting the growth of phytopathogens. Over a long period of co-evolution, endophytic microbes have attained the mechanism of synthesis of various hydrolytic enzymes such as pectinase, xylanases, cellulase, and proteinagro-environmental sustainability.Purpose Risk to healthcare workers treating asymptomatic patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the operating room depends on multiple factors. This review examines the evidence for asymptomatic or pre-symptomatic carriage of SARS-CoV-2, the risk of transmission from asymptomatic patients, and the specific risks associated with aerosol-generating procedures. Protective measures, such as minimization of aerosols and use of personal protective equipment in the setting of treating asymptomatic patients, are also reviewed. Source We examined the published literature as well as Societal guidelines. Principal findings There is evidence that a proportion of those infected with SARS-CoV-2 have detectable viral loads prior to exhibiting symptoms, or without ever developing symptoms. The degree of risk of transmission from asymptomatic patients to healthcare providers will depend on the prevalence of disease in the population, which is difficult to assess without widespread population screening. https://www.selleckchem.com/ Aerosol-generating procedures increase the odds of viral transmission from infected symptomatic patients to healthcare providers, but transmission from asymptomatic patients has not been reported. Techniques to minimize aerosolization and appropriate personal protective equipment may help reduce the risk to healthcare workers in the operating room. Some societal guidelines recommend the use of airborne precautions during aerosol-generating procedures on asymptomatic patients during the coronavirus disease pandemic, although evidence supporting this practice is limited. Conclusion Viral transmission from patients exhibiting no symptoms in the operating room is plausible and efforts to reduce risk to healthcare providers include reducing aerosolization and wearing appropriate personal protective equipment, the feasibility of which will vary based on geographic risk and equipment availability.Objective The present study aimed to identify and quantify the elution of monomers of five different resin-based CAD/CAM blocks (RCBs) using HPLC. Methods Five different RCBs were used in the study GC Cerasmart (CS), Voco Grandio blocs (GR), 3M Lava Ultimate (LU), Shofu Block (SB), and Vita Enamic (VE). Fifteen samples from each material were prepared using a low-speed precision diamond saw (ISOMET Buehler, Lake Bluff, IL, USA) at 5 × 5 × 4 mm size. After the preparation of samples, an extraction solution was mixed with %75/%25 ethanol/water. The samples were stored in the amber-colored bottles during three different immersed periods as 1 h, 24 h, and 90 days (n = 5). After immersion, 0.5 ml solutions were taken from each bottle and analyzed using HPLC. Results A total of 16.7 μg/ml of monomers from SB, 13.4 μg/ml of monomers from GR, 13.2 μg/ml of monomers from CS, and 6.7 μg/ml of monomers from LU were found after 3-m immersion. TEGDMA after 3-m of immersion was only released from the SB group, and also BisEMA was released from the CS group. Among the specimens immersed for 1 h, UDMA was released the least from the LU group and the most from the GR group (p less then 0.05). Correspondingly, 24 h and 3 m after immersion, the highest release of monomers was found in the GR (p less then 0.05). Conclusions When the monomer release from RCBs was evaluated, it was shown that these materials released methacrylate-based monomers except VE, especially if they were kept in a solvent solution for a long time such as 3 m. Clinical significance The novel resin-based CAD/CAM blocks might monomer release, which may cause cytotoxic effects. But, the detected amount of monomer release is below the estimated daily limit.

Among top ranked research-intensive Schools of Nursing, awards to non-nurse Principal Investigators (PIs) averaged 34.5% across all years. The percentage of NIH funds awarded to non-nurse PIs ranged from 0% at 3 Schools to as high as 97% at 1 School of Nursing. Over the past 5 years, the following Institutes have consistently been the largest funders (total dollars) to Schools of Nursing NINR, National Institute of Aging, National Institute of Minority and Health Disparities, National Cancer Institute and National Institute of Child Health and Human Development. CONCLUSION These findings highlight the current funding streams for Schools of Nursing as well as opportunities for expansion. Preparing a cadre of nurse scientists who can generate new knowledge to advance our health care is critical to the success of our profession and to ensuring the health of the people for whom we provide care. As the demographics in the United States continue to change, nurses must deliver care to patients from diverse cultural backgrounds. Cultural humility is a lifelong process of self-reflection which is also defined by that individual. It allows an individual to be open to other people's identities, which is core to the nursing standard of providing holistic care. Embracing and incorporating cultural humility is essential for creating a comprehensive and individualized plan of care. One of the ways to achieve cultural humility in nursing is to train future faculty to become agents of cultural humility. This also helps to create a pipeline of nurses who have respect and empathy for the patients they serve. The aims of this paper include 1) define cultural humility and its importance to healthcare professionals; 2) explore the intrapersonal, interpersonal, and system levels of cultural humility; 3) provide insight on how to promote cultural humility; 4) reflect on best practices across a variety of healthcare disciplines; and 5) provide suggestions for practice. BACKGROUND Nursing students from historically underrepresented ethnic minorities and disadvantaged background (HUREM-DB) groups often face barriers such as a lack of consistent financial resources, fewer professional role models, bias, and micro-inequities. Utilizing a multifaceted approach for support can be crucial to enhancing student success. PURPOSE OF THE PROJECT MENTORS2 mitigates some of the challenges for HUREM-DB nursing students with educational, cultural, social, and financial resources. Courageous dialogue (CD) was one required activity of MENTORS2 and included topics such as stress management, time management, and honors project preparation. IMPLEMENTATION OF THE PROJECT Courageous dialogue sessions were conducted with 56 HUREM-DB undergraduate nursing students enrolled in a baccalaureate program. The number of evaluations submitted for a session averaged 17 (range 7-36). Courageous dialogue sessions allowed students to express views in a safe environment with opportunities for peer support, role modeling, open discussion, and problem solving. PROJECT OUTCOMES Student evaluations reflected an appreciation of the opportunity to share experiences and learn new skills, knowledge, and approaches to aid their success in nursing school and perhaps their entry into the profession of nursing. CONCLUSION Courageous dialogue can be an important part of a comprehensive strategy to support HUREM-DB nursing students academically, socially, and professionally. BACKGROUND A racially diverse registered nurse workforce could improve health outcomes for minority populations, and efforts to recruit students from underrepresented groups need to be strengthened. PURPOSE To explore the perceptions of nursing held by African American undergraduate non-nursing science majors within the context of their career ideals. METHOD Qualitative description was the research design utilized in this study. A purposive sample of 20 African American non-nursing science majors participated in semi-structured interviews. Miles, Huberman, and Saldaña's (2014) coding processes for qualitative analysis were employed. RESULTS Three themes were identified that described career behavior, ideals, and the participants' perceptions of nursing as a career 1.) Career Development Internal and External Contributions, 2.) The Ideal Career Profession with Power, and 3.) Perception of Nursing A Limited Profession. Findings suggest African American science majors carry a significant burden to overcome stereotypes for themselves and their communities, thus desire careers with power and autonomy. CONCLUSION Careers in medicine were idealized whereas careers in nursing were devalued. These college students often embraced careers in medicine due to, in part, the perceived ability to overcome stereotypes and simultaneously rejected nursing careers due to desire to avoid negatively stereotyped careers. OBJECTIVE This study investigated the effect of plasma-treated leno weaved ultra-high-molecular-weight polyethylene fiber placement on gap formation and microtensile bond strength (MTBS) of a bulk-fill composite in deep cavity. METHODS Resin composite molds (3 mm width, 4 mm depth) were treated with Clearfil SE Bond 2 and restored with 3 techniques (1) Surefil SDR flow (SDR) placed in bulk (BLK), (2) SDR placed in two unequal increments (INC) and (3) SDR placed after an increment of SDR placed with wetted polyethylene fiber (Ribbond Ultra) at the cavity floor (FRC). As a control, the cavities were bulk-filled with SDR and no bonding agent (n = 12). All the specimens were subjected to real-time and 3D imaging by SS-OCT (1330 nm) to calculate the total volume of gap formed (mm3) at the cavity floor and between the composite increments. For MTBS, the occlusal cavities of the similar dimensions (3 × 3 × 4 mm3) were prepared on extracted molars with similar composite placement techniques (BLK, INC and FRC). Aftereinforced increment acts as a shrinkage stress breaker and protects the bonded interface at deep dentin. https://www.selleckchem.com/products/VX-770.html BACKGROUND Molecular pathogenesis of Triple-negative breast cancer (TNBC) is inconclusively documented from resource limited countries and hence there is a lack of available targeted therapy for clinical interventions. Compared to other breast cancer subtypes, TNBC is more aggressive, higher recurrence rate, and higher prevalence in younger premenopausal women. Sporadic literature indicates predominance of TNBC in all reported breast cancer cases from Northeast India. AIM This study was conducted to evaluate the candidature of panel of key molecular markers involved in the development and progression of TNBC for prognosis and futuristic tailored targeted therapy. MATERIALS AND METHODS We analyzed the clinicopathological characterized and immunohistochemically screened the differential expression of key molecular markers involved in the development and progression of in TNBC cases vis-a-vis non-TNBC and autopsy-based control samples. RESULTS TNBC tends to display at an early reproductive age and is more aggressive in nature.

In this manuscript, we report the development of a rapid and facile optic sensor for highly sensitive and selective detection of cobalt ions (Co(II)). The detection strategy was based on the specific silver nanoparticle-glutathione interaction and later on secondary interaction of this structure with cysteine and Co(II) ion. The interaction of these structures creates a new absorption band in the UV region. The intensity change of this band can be correlated by Co(II) ion concentration. The addition of cysteine decreases the Surface Plasmon Resonance (SPR) of silver nanoparticles but does not provide quantitative information. The proposed method offers the advantage of improved sensitivity for detection of Co(II) ions in a very short time period. Co(II) ions create a unique absorption peak during the complex formation and this peak provides sensitive determination of this metal ion in existence of other metal ions. The peak is visible only after taking the first derivative of absorption spectra. Under the optimized conditions, the detection limit of the method is around 0.68 μM. In addition, the synthesized silver nanoparticles (AgNPs) were characterized by atomic force microscopy (AFM). The proposed metal ion sensor provides a very facile and convenient way to determine the concentration of Co(II) ions in aqueous system.Dew retting of fiber crops, such as hemp or flax, in the field after harvest promotes the microbial biodegradation of the tissues surrounding cellulosic fibers, which helps preserve the quality of fibers during their extraction and valorization for industry. This bioprocess is currently the bottleneck for plant fiber valorization because it is empirically managed and its controlling factors have not been properly quantified. A novel multiscale model representing tissue and polymer biodegradation was developed to simulate microbial growth on the stem during retting. The model was evaluated against experimental hemp retting data. It consistently simulated the mass loss of eight plant polymers belonging to two tissues of the stem outer layer, i.e., parenchyma and fiber bundles. Microbial growth was modeled by Monod equations and modulated by the functions of temperature and moisture. This work provides a tool for gaining more insights into microorganism behavior during retting under local climate conditions.Cytochrome P450 OleT is a fatty acid decarboxylase that uses hydrogen peroxide (H2O2) to catalyze the production of terminal alkenes, which are industrially important chemicals with biofuel and synthetic applications. Despite its requirement for large turnover levels, high concentrations of H2O2 may cause heme group degradation, diminishing enzymatic activity and limiting broad application for synthesis. Here, we report an artificial enzyme cascade composed of glucose oxidase (GOx) and OleTSA from Staphylococcus aureus for efficient terminal alkene production. By adjusting the ratio of GOx to OleTSA, the GOx-based tandem catalysis shows significantly improved product yield compared to the H2O2 injection method. Moreover, the co-assembly of the GOx/OleTSA enzymes with a polymer, forming polymer-dual enzymes nanoparticles, displays improved activity compared to the free enzyme. This dual strategy provides a simple and efficient system to transform a naturally abundant feedstock to industrially important chemicals.The gas-liquid mass transfer coefficient (kLa) of O2 was investigated in a bubble column reactor (BCR) using a sintered gas filter (SF), ceramic membrane module (CMM), and hollow fiber membrane module (HFM), which have different ranges of gas supply areas. kLa was enhanced by increasing flow rate in all of the spargers. Different responses when changing the gas supply area were obtained depending on the sparger type. Average values of kLa that were 52 and 258% higher were obtained using a CMM-integrated BCR compared to SFs and HFMs. CO-water kLa was investigated using CMMs for application to gas fermentation. The CO-water kLa ranged from 28.3 to 113.7/h under the experimental conditions. Based on the experimental data from CO and O2, a model to predict kLa was constructed for CMM-integrated BCRs. A dimensionless number indicating a gas supply area of the sparger was newly defined and included in the developed model.Per- and polyfluoroalkyl substances (PFAS) are a class of persistent chemicals used as industrial surfactants, fire-fighting foams, and textile treatments. Early childhood exposure to perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), perfluorononanoic acid (PFNA), and perfluorohexane sulfonic acid (PFHxS) may affect the immune system to increase the risk of allergic and respiratory diseases. However, there are substantial gaps in our knowledge about the relationship between PFAS and immune-mediated outcomes such as asthma in children. Thus, we examined the cross-sectional associations of serum PFOA, PFOS, PFNA, and PFHxS concentrations with childhood asthma. We used data from children aged 3-11 years who participated in the National Health and Nutrition Examination Survey (2013-2014). Serum PFAS concentrations were measured in serum using analytical chemistry methods. Asthma was assessed by parent-reported, doctor-diagnosed, asthma using a standardized questionnaire. Controlling for covariates, we estimated odds ratios for asthma per standard deviation increase in ln-transformed serum PFAS concentrations (n = 607). We also examined effect measure modification by child age, sex, and race/ethnicity. PFOA (1.1; 95% CI 0.8, 1.4), PFOS (1.2; 95% CI 0.8, 1.7), PFNA (1.1; 95% CI 0.8, 1.6), and PFHxS (1.1; 95% CI 0.9, 1.6) were weakly associated with an increased odds of asthma. Age modified associations between serum PFOS, but not other serum PFAS concentrations, and odds of asthma (age x PFOS interaction term p-value = 0.03). https://www.selleckchem.com/products/beta-glycerophosphate-sodium-salt-hydrate.html Sex and race/ethnicity did not modify these associations. We observed some evidence that serum PFAS concentrations are weakly associated with increased asthma prevalence in US children.Exposure to perfluoroalkyl substances (PFAS) was found to be associated with several pathological endpoints, including high cholesterol levels, specific defective functions of the immune system and reduced birth weight. While environmental PFAS have been recognized as threats for public health, surprisingly little is known about the underlying mechanisms of toxicity. We hypothesized that some of the observed vascular and developmental effects of environmental PFAS may share a common molecular pathway. At elevated levels of exposure to PFAS, a reduction in mean birth weight of newborns has been observed in combination with a high incidence rate of preeclampsia. As both, preeclampsia and reduced birth weight are consequences of an inadequate placental vascularization, we hypothesized that the adaptation of placental vasculature may get compromised by PFAS. We analyzed pseudo-vascular network formation and protein expression in the HTR8/SVneo cell line, an embryonic trophoblast cell type that is able to form vessel-like vascular networks in 3D-matrices, similar to endothelial cells.

Introduction The off periods in Parkinson's disease have a significantly negative impact on quality of life. What the most bothersome aspects of off periods are from the patient's perspective are not well studied, nor is the degree to which screening tools for wearing off such as the Wearing Off Questionnaires (WOQs) capture what bothers patients most. Methods A questionnaire was deployed to eligible participants of Fox Insight, an online study of individuals with self-reported Parkinson's disease. Inclusion criteria were the use of ≥1 dopaminergic medications and an affirmative response to a question on experiencing off periods. Participants provided free-text responses regarding the top 3 most bothersome symptoms they experience when off. A determination was made regarding whether each response would have been captured by the 32-item, 19-item, and 9-item WOQs. Results The final sample had 2106 participants, a mean age of 66.6 years, 52.3% were men, and had a disease duration of 4.9 years. The WOQ-32 items covered all of the most bothersome symptoms for 53.2% of respondents. Among bothersome aspects of off not captured by the WOQs, 597 (66.2%) were specific symptoms, with freezing of gait, apathy, and memory problems being the most common. The functional consequences of off periods were most bothersome to 232 (25.7%), with walking problems being the most common. The emotional response to off periods was the most bothersome aspect to 169 respondents (18.7%). Discussion This study emphasizes the value of narrative data in understanding patient experiences, and what bothers patients most about off periods. The WOQs, although of established utility in the screening for wearing off, may not capture those symptoms most bothersome to patients. © 2020 International Parkinson and Movement Disorder Society.Background Few studies assess the relationships between nonmotor aspects of experiences of daily living and cognitive functioning in Parkinson's disease (PD). Objective To evaluate the relationships among the Movement Disorders Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS) part I items and neuropsychological tests in PD.Methods We assessed 151 PD patients with the MDS-UPDRS part I and a battery of cognitive tests focused on the following 5 cognitive domains attention/working memory, executive functioning, recent memory, language, visuoperception. Raw scores for individual cognitive tests were transformed to z scores, and cognitive domain scores were calculated by averaging z scores within each domain. Individual items from the MDS-UPDRS part I were entered in a stepwise linear regression analysis assessing item contribution to cognitive domain scores. Results The MDS-UPDRS part I item scores for hallucinations and psychosis and light headedness on standing predicted attention/working memory domain scores (P = 0.004). These same item scores, along with apathy, depressed mood, and dopamine dysregulation syndrome, predicted executive functioning (P = 0.044). The apathy and dopamine dysregulation syndrome items predicted language (P = 0.006). In addition, the cognitive impairment and sleep items were predictors of recent memory (P = 0.031). None of the items were predictors of visuoperception (P = 0.006). Other part I items were not significantly related to cognitive domain scores. Conclusions Specific nonmotor MDS-UPDRS part I items, particularly mood, behavior, and autonomic-related items, exhibited significant relationships with cognitive domains. The highest number of items were predictive of the executive functioning domain, which is the hallmark cognitive dysfunction in PD. © 2020 International Parkinson and Movement Disorder Society.Background and Objective The delayed-on phenomenon (DOP) related to levodopa treatment frequently disturbs quality of life in advanced-stage Parkinson's disease (PD) patients. The objective of this study was to explore the impact of swallowing dysfunction on the development of DOP. Methods Swallowing function was investigated by endoscopic evaluation in 11 PD patients with the DOP and 9 PD patients without the DOP during the on phase. Residual drug in the pharynx after taking the drug in tablet, capsule, and powder forms was also observed. Results Residual drug was seen in the pharynx in six cases (30.0%). Pooling of saliva, delayed swallowing reflex, and residual drug were more frequent in the DOP group than in the group without the DOP (P  less then  0.05). https://www.selleckchem.com/products/way-262611.html The odds ratios for residual drug in the pharynx, pooling of saliva, and delayed swallowing reflex for the DOP were 42.7 (95% confidence interval, 1.89-962.9), 14.0 (95% confidence interval, 1.25-156.6), and 15.8 (95% confidence interval, 1.75-141.4), respectively. Conclusions These results suggest that swallowing dysfunction leading to residual antiparkinsonian drug in the pharynx has substantial impacts on the DOP in PD patients. © 2020 International Parkinson and Movement Disorder Society.Background Corticobasal degeneration (CBD) can present with various clinical phenotypes including Richardson's syndrome (RS). Although neuropathological examination can differentiate CBD and progressive supranuclear palsy (PSP) pathologies, no clinical or imaging findings can differentiate CBD from other pathologies when a patient presents with a variant type of CBD. As these various phenotypes are associated with non-CBD pathologies, clinical diagnostic accuracy can be low for such patients. Objectives To present clinical features of two cases with symptom progression in line with PSP-RS, who were diagnosed with CBD based on neuropathological examination. Methods Baseline, follow up examinations, and detailed neuropathological examinations of two CBD cases presenting and progressing in line with probable PSP-RS are demonstrated. Results The two cases clinically diagnosed as probable PSP-RS were shown to have CBD upon neuropathological examination, which is the gold standard for diagnosis of both PSP and CBD. Conclusions These cases emphasize the importance of neuropathology for the definite diagnosis, and stress the need for distinctive markers to increase the reliability of clinical diagnosis before death. © 2020 International Parkinson and Movement Disorder Society.

Background Past studies have shown that elevated estradiol levels could inhibit penile erection, but the relationship between estradiol and erection of the penile tip or base has not been extensively researched. Methods We therefore investigated estradiol's effects on the erection of the penile tip and base, with a cross-sectional study of 135 patients with erectile dysfunction (ED), based on scores of ≤21 according to the International Index of Erectile Function-5. All patients were tested for nocturnal penile tumescence, blood pressure (BP), serum glucose, total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), progesterone (P), estradiol (E), and testosterone (T). Univariate and multivariate analyses were used to assess associations between estradiol levels and erection at the penile tip and base. Results We found no obvious relationship between erection time at penile tip and estradiol levels but did observe a negative correlation between base erection time and estradiol level [hazard ratio (HR) ‒0.11; 95% CI ‒0.80-1.72]. With increasing estradiol concentration, multivariate analysis showed an obvious reduction in base erection time among patients with normal Rigiscan results (HR ‒0.31; 95% CI ‒1.63-1.29) (P less then 0.05) as estradiol concentration increased. Conclusions Our data indicate that estradiol inhibits penile erection, particularly at the penile base. Also, when the effective erection time of the penile base lasts longer than 10 min, estradiol has a more obvious inhibitory effect on penile base erection. 2019 Translational Andrology and Urology. All rights reserved.Out of hospital cardiac arrest (OHCA) is a leading cause of death worldwide. Early cardiopulmonary resuscitation (CPR) and early defibrillation are key to improving outcomes of patients with OHCA including return of spontaneous circulation (ROSC) and survival to hospital discharge with good neurologic outcomes. Lebanon like other developing countries, suffers from absence of organized prehospital cardiac arrest care bundle and from absence of a legal framework for community involvement in cardiac arrest care. Scientific societies, involved non-governmental organizations (NGOs) and local governmental stakeholders organized a national meeting to launch a strategy aiming at improving OHCA outcomes in Lebanon. This article represents a position statement of the Lebanese Society of Cardiology and the Lebanese Society of Emergency Medicine summarizing the strategy to improve out-of-hospital CPR. Participating stakeholders developed and submitted a law proposal of a "Good Samaritan Law" to the Lebanese parliament. Several of activities were also launched aiming at establishing public access defibrillation programs and at training bystanders in different areas in Lebanon to perform bystander CPR and use automated external defibrillators (AEDs). Additional recommendations were proposed to local emergency medical system (EMS) agencies to improve prehospital care and introduce medical direction to prehospital activities. 2019 Cardiovascular Diagnosis and Therapy. All rights reserved.A patient presented with an intraluminal fibrous cord in the distal segment of ascending aorta. On axial images, the fibrous cord had the appearance of an aortic dissection flap. Observation of continuous images and multiplanar reconstruction were critical for differential diagnosis. Volume render images showed both ends of the fibrous cord had tentiform attachments to the aorta inner wall, which indicating a congenital aorta anomaly. One possible hypothesis for the congenital variation might be the fifth primitive aortic arches remnant. 2019 Cardiovascular Diagnosis and Therapy. All rights reserved.Background Little is known about the long-term outcomes of medical therapy (MT) versus successful percutaneous coronary intervention (PCI) in elderly patients with coronary chronic total occlusions (CTOs). Methods There were 1,294 consecutive patients with 1,520 CTOs included (2007 to 2016) and were divided into the younger group (age less then 65 years; n=664, 51.3%) and the older group (age ≥65 years; n=630, 48.7%). In the older group, 630 patients were divided into MT group (n=421) and successful CTO-PCI group (n=209) according to the initial treatment strategy. In the younger group, they were divided into two groups 379 patients in the MT group and 285 patients in the successful CTO-PCI group. We performed propensity score matching to minimize any selection bias. The primary end point was cardiac mortality. The secondary end point was major adverse cardiac event (MACE). Results After 3.6 (IQR, 2.1-5.0) years follow-up, no significant difference was observed between the MT and successful CTO-PCI groups inardiovascular Diagnosis and Therapy. All rights reserved.Background Left ventricular (LV) extracellular volume fraction (ECV) provides prognostic information in patients with variety of cardiomyopathies. However, data on the clinical significance of LV ECV in patients with atrial fibrillation (AF), especially in patients without replacement fibrosis are sparse. This study sought to investigate whether the presence of LV fibrosis identified by cardiac magnetic resonance (CMR) ECV quantification would independently predict the recurrence of AF after first catheter ablation (CA) in patients with AF. Methods A total of 130 consecutive patients who were referred for CA of AF underwent CMR examination prior to ablation. LV function, T1 mapping derived LV ECV, LV late gadolinium enhancement (LGE) were assessed. Patients were followed for arrhythmia recurrence after the CA procedure. Results Of 130 AF patients, 65 patients had paroxysmal AF, and 65 patients had persistent AF. There were 50 AF recurrences over a median follow-up period of 13 months. LV ECV were significantly higher in patients with recurrent AF compared to those with no recurrence (30.4%±3.3% vs. 27.4%±2.9%, P less then 0.001). In multivariable model, gender (HR 0.348, 95% CI 0.174-0.697, P=0.003), body mass index (BMI) (HR 1.159, 95% CI 1.050-1.279, P=0.003), AF duration (HR 1.006, 95% CI 1.001-1.011, P=0.017), and LV ECV (HR 1.158, 95% CI 1.071-1.251, P=0.000) were significantly associated with AF recurrence. https://www.selleckchem.com/products/mevastatin.html In subgroup of patients without LGE, gender, BMI, AF duration and LV ECV were still the independent predictors of AF recurrence. Conclusions LV ECV expansion is associated with AF recurrence after CA and is a strong independent predictor of AF recurrence. 2019 Cardiovascular Diagnosis and Therapy. All rights reserved.

Standard approaches for identifying task-completion strategies, such as precrastination and procrastination, reduce behavior to single markers that oversimplify the process of task completion. To illustrate this point, we consider three task-completion strategies and introduce a new method to identify their use. This approach was tested using an archival data set (N = 8,655) of the available electronic records of research participation at Kansas State University. The approach outperformed standard diagnostic approaches and yielded an interesting finding Several strategies were associated with negative outcomes. Specifically, both procrastinators and precrastinators struggled to finish tasks on time. Together, these findings underscore the importance of using holistic approaches to determine the relationship among task characteristics, individual differences, and task completion.Objective Studies conducted with Latina/o populations suggest there are gendered norms surrounding drinking behavior, but research examining alcohol use among Latina college students in the context of traditional feminine gender role (TFGR) norms, is scarce.Participants and Methods 405 Latina undergraduates completed a web-based study of sociocultural factors and alcohol use. Path analyses assessed the relation between TFGR , alcohol use, and alcohol-related protective behavioral strategies (PBS).Results A dimension of TFGR characterized by purity was related to (a) reduced alcohol use in general, and (b) PBS designed to modify the manner of drinking among drinkers. In turn, this dimension of PBS related to a lower likelihood of experiencing drunkenness.Conclusions These findings support the notion that enacting some dimensions of TFGR among Latina college students may limit general alcohol use (directly) and high-risk drinking (indirectly through use of PBS), while enacting other dimensions may place Latinas at risk of alcohol misuse.Objective The psychometric properties of a measure of social determinants of health for college students entitled, The LIFESCREEN-C was developed. Participants A sample of 226 college students completing an online survey during the 2018-2019 academic year. Methods Tetrachoric correlations were used to confirm a three factor model. Results Results found model fit; convergent validity with a measure of general health; and adequate internal reliability. Conclusions The three model fit included general social health needs, college student social health needs, and promotive social health factors. Implications for health professionals in college settings concludes.Why do some cultural items succeed and others fail? Some scholars have argued that one function of the narrative arts is to facilitate feelings of social connection. If this is true, cultural items that activate personal connections should be more successful. The present research tested this possibility in the context of second-person pronouns. We argue that rather than directly addressing the audience, communicating norms, or encouraging perspective taking, second-person pronouns can encourage audiences to think of someone in their own lives. Textual analysis of songs ranked in the Billboard charts (N = 4,200), as well as controlled experiments (total N = 2,921), support this possibility, demonstrating that cultural items that use more second-person pronouns are liked and purchased more. These findings demonstrate a novel way in which second-person pronouns make meaning, how pronouns' situated use (object case vs. subject case) may shape this meaning, and how psychological factors shape the success of narrative arts.In the spring of 2018, a 10-day field study was conducted in Colorado's Denver-Julesburg oil and natural gas production basin to improve information on well pad pneumatic controller (PC) populations and identify PCs with potential maintenance issues (MIs) causing excess emissions through a novel optical gas imaging (OGI) survey approach. https://www.selleckchem.com/products/way-262611.html A total of 500 natural gas-emitting PCs servicing 102 wells (4.9 PCs/well) were surveyed at 31 facilities operated by seven different companies. The PCs were characterized by their designed operational function and applications, with 83% of the PC population identified as intermittent PCs (IPCs). An OGI inspection protocol was used to investigate emissions on 447 working PCs from this set. OGI detected continuous emissions from 11.3% of observed IPCs and these were classified as experiencing some level of MIs. OGI imaging modes were observed to have a significant effect on emission detectability with high sensitivity mode detection rates being approximately 2 times higher comand supporting emission measurements, was demonstrated as an effective method to identify controllers with potential maintenance issues causing excess emissions. The results of the pneumatic controller and optical gas imaging surveys improved information on pneumatic controller populations within the basin and also demonstrated the significant effect optical gas imaging modes have on emission detections.Objective This study examined trends in functionally impairing depression, significant anxiety, suicidal ideation, and suicide attempts among college student-athletes across time, racial/ethnic group, and student-athlete standing.Participants The sample consisted of 39,840 White, Black, Hispanic/Latinx, Asian/Pacific Islander, Native American, and Multiracial students who reported participating in "varsity" level college athletics in the past 12 months during the Spring 2010-2015 administrations of the American College of Health Association National College Health Assessment II.Methods Chi-square and difference in proportion tests compared mental health rates across time, racial/ethnic groups, and student-athlete status (student-athlete versus non-athlete).Results Findings suggested a lack of differences in mental health rates between 2010 and 2015, with the exception of a higher rate of anxiety for White student-athletes in 2015. Racial/ethnic comparisons pointed to relatively consistent heightened risks for depression and suicide concerns among Asian/Pacific Islander and Multiracial student-athletes compared to White student-athletes. Student-athlete status appeared most consistently to benefit White student-athletes, whereas there was evidence of potential liability of student-athlete status for Asian/Pacific Islander student-athletes.Conclusions Results make the case for adopting a racially/ethnically aware cultural lens when conceptualizing mental health risk and resilience among student-athletes.

d Novoa.Somatic cell nuclear transfer (SCNT) has broad applications but is limited by low cloning efficiency. In this review, we mainly focus on SCNT-mediated epigenetic reprogramming in livestock and also describe mice data for reference. This review presents the factors contributing to low cloning efficiency, demonstrates that incomplete epigenetic reprogramming leads to the low developmental potential of cloned embryos, and further describes the regulation of epigenetic reprogramming by long non-coding RNAs, which is a new research perspective in the field of SCNT-mediated epigenetic reprogramming. In conclusion, this review provides new insights into the epigenetic regulatory mechanism during SCNT-mediated nuclear reprogramming, which could have great implications for improving cloning efficiency. Copyright © 2020 Wang, Qu, Li, He, Liu and Huan.Polyploidization often leads to "transcriptome shock," and is considered an important factor in evolution of species. Analysis of the cell cycle, which is associated with survival in polyploidy, has proved useful in investigating polyploidization. Here, we used mRNA sequencing to investigate global expression in vitro (in cultured cells) and in vivo (in fin and liver tissues) in both the diploid and tetraploid Carassius auratus red var.. Differential expression (DE) of genes in diploid (7482, 36.0%) and tetraploid (3787, 18.2%) states suggested that in vitro and in vivo conditions dramatically change mRNA expression levels. However, of the 20,771 total shared expressed genes, 18,050 (87.0%), including 17,905 (86.2%) non-differentially expressed genes (DEGs) and 145 (0.7%) DEGs between diploids and tetraploids, showed the same expression trends in both cultured cells and liver tissues. Of the DEGs, four of seven genes in the cell cycle pathway had the same expression trends (upregulated in diploids and tetraploids) in both cultured cells and liver tissues. Quantitative PCR analysis confirmed the same expression trends in the nine DEGs associated with regulation of the cell cycle. This research on common characteristics between diploids and tetraploids provides insights into the potential molecular regulatory mechanisms of polyploidization. The steady changes that occur between diploids and tetraploids in vitro and in vivo show the potential value of studying polyploidy processes using cultured cell lines, especially with respect to cell cycle regulation. Copyright © 2020 Ren, Lu, Fan, Hu, Li, Xiao and Liu.Rationale Mucopolysaccharidosis type II (Hunter syndrome) is an X-linked multisystem disorder, caused by deficiency of the lysosomal enzyme iduronate-2-sulfatase (I2S). The clinical manifestations of this disease are severe skeletal deformities, airway obstruction, cardiomyopathy, and neurologic deterioration. Patient The patient was 5 years and 6 months boy, with developmental delay, hearing loss, hepatosplenomegaly, and skeletal dysplasia. He was diagnosed with mucopolysaccharidosis type II based on clinical manifestations, biochemical and genetic analysis. Outcomes The patient carries a new mutation (c.879-1210_1007-218del) in hemizygosis in the IDS gene, which was defined as pathogenic according to the 2015 American College of Medical Genetics and Genomics-Association for Molecular Pathology guidelines and as responsible for the mucopolysaccharidosis type II phenotype in the patient. Copyright © 2020 Gomes, Marins, Motta, Kyosen, Curiati, D’Almeida, Martins and Pesquero.[This corrects the article DOI 10.3389/fpls.2020.00115.]. Copyright © 2020 Han, Lu, Guo, Qiao, Sui, Qiu and Wang.Chromosomal inversions have the potential to play an important role in evolution by reducing recombination between favorable combinations of alleles. Until recently, however, most evidence for their likely importance derived from dipteran flies, whose giant larval salivary chromosomes aided early cytogenetic studies. https://www.selleckchem.com/products/mevastatin.html The widespread application of new genomic technologies has revealed that inversions are ubiquitous across much of the plant and animal kingdoms. Here we review the rapidly accumulating literature on inversions in the plant kingdom and discuss what we have learned about their establishment and likely evolutionary role. We show that inversions are prevalent across a wide range of plant groups. We find that inversions are often associated with locally favored traits, as well as with traits that contribute to assortative mating, suggesting that they may be key to adaptation and speciation in the face of gene flow. We also discuss the role of inversions in sex chromosome formation, and explore possible parallels with inversion establishment on autosomes. The identification of inversion origins, as well as the causal variants within them, will advance our understanding of chromosomal evolution in plants. Copyright © 2020 Huang and Rieseberg.Triploid breeding is a central way to improve growth traits, timber quality, and stress resistance in Populus. In the present study, the morphology and viability of colchicine-induced 2n pollen, triploid production by crossing induced 2n pollen, and identification of genetic constitution of colchicine-induced 2n pollen were conducted in Populus canescens based on optimizing technology for inducing chromosome doubling in pollen. We found that the meiotic stage, injection time, and the interaction between the meiotic stage and injection time had highly significant effects on the 2n pollen production rate. The most effective treatment for inducing 2n pollen was to give 11 injections of 0.5% colchicine solution when pollen mother cells (PMCs) were at the pachytene stage. The highest 2n pollen production rate was 30.27 ± 8.69%. Colchicine occasionally affected ectexine deposition, and some narrow furrows were detected in the ectexine structure. However, no significant difference was observed in the pollen germination rate between natural 2n pollen and colchicine-induced 2n pollen. Moreover, 5 triploids derived from FDR-type 2n pollen were generated by crossing induced 2n pollen, suggesting that colchicine does not eliminate the function of colchicine-induced 2n pollen. However, slower growth of 2n pollen tubes was responsible for a lower triploid production rate. Copyright © 2020 Zhou, Wu, Sang, Zhao, Zhang and Liu.