Soy whey and brewer's spent grain hydrolysates wholly replace conventional medium for microalgae growth: Process performance and economic considerations.

To enhance circularity in heterotrophic microalgal bioprocesses, this study completely substituted glucose and Bold's basal medium (BBM) with brewer's spent grain (BSG) and soy whey (SW) hydrolysates. Mild acid hydrolysis conditions of BSG (0.2 M H2SO4, 130 °C, 36 min) and SW (0.1 M HCl, 95 °C, 30 min) were optimised for glucose release, and their hydrolysates were optimally mixed (15 % SW-85 % BSG) to obtain a medium that best supported Auxenochlorella protothecoides growth. Maximum biomass production (Xmax) and productivity (PXmax) obtained in the hydrolysate medium containing 50.75 g/L endogenous glucose (Xmax: 22.17 g/L; PXmax: 7.06 g/L/day) were comparable to that in BBM containing 50.44 g/L exogenous glucose (Xmax: 20.02 g/L; PXmax: 6.34 g/L/day). Moreover, estimated hydrolysate medium production costs were within an order of magnitude to BBM. Overall, the integrated approach of tailored hydrolytic treatments and complementary side-streams presents a promising technical and economic feasibility, with applications extending beyond A. protothecoides.

Achieving efficient and stable blue thermally activated delayed fluorescence organic light-emitting diodes based on four-coordinate fluoroboron emitters by simple substitution molecular engineering.

Achieving both high efficiency and high stability in blue thermally activated delayed fluorescence organic light-emitting diodes (TADF-OLEDs) is challenging for practical displays and lighting. Here, we have successfully developed a series of sky-blue to pure-blue emitting donor-acceptor (D-A) type TADF materials featuring a four-coordinated boron with 2,2'-(pyridine-2,6-diyl)diphenolate (dppy) ligands, i.e.1-8. Synergistic engineering of substituents on the phenyl bridge as well as the electronic properties and the attached positions of heteroatom N-donors not only enables fine-tuning of the emission colors, but also modulates the nature and energies of their triplet excited states that are important for the reverse intersystem crossing (RISC). Particularly for the compound with two methyl substituents on the phenyl bridge (compound 8), RISC is significantly facilitated through the vibronic coupling of the energetically close-lying triplet charge transfer (3CT) and the triplet local excited (3LE) states, when compared to analogue 7. Efficient sky-blue to pure-blue OLEDs with electroluminescence peaks (λ EL) at 460-492 nm have been obtained, in which ca. five-fold higher external quantum efficiencies (EQEs) of 18.9% have been demonstrated by 8 than that by 7. Moreover, ca. thirty times longer device operational half-lifetimes (LT50) of 9113 hours for 8 than that for 7 as well as satisfactory LT50 reaching 26 643 hours for 6 at an initial luminance of 100 cd m-2 have also been demonstrated. To the best of our knowledge, these results represent one of the best high-performance blue OLEDs based on tetracoordinated boron TADF emitters. Moreover, the design strategy presented here has provided an attractive strategy for enhancing the device performance of blue TADF-OLEDs.

Application of image recognition-based tracker-less augmented reality navigation system in a series of sawbone trials.

BACKGROUND: This study introduced an Augmented Reality (AR) navigation system to address limitations in conventional high tibial osteotomy (HTO). The objective was to enhance precision and efficiency in HTO procedures, overcoming challenges such as inconsistent postoperative alignment and potential neurovascular damage. METHODS: The AR-MR (Mixed Reality) navigation system, comprising HoloLens, Unity Engine, and Vuforia software, was employed for pre-clinical trials using tibial sawbone models. CT images generated 3D anatomical models, projected via HoloLens, allowing surgeons to interact through intuitive hand gestures. The critical procedure of target tracking, essential for aligning virtual and real objects, was facilitated by Vuforia's feature detection algorithm. RESULTS: In trials, the AR-MR system demonstrated significant reductions in both preoperative planning and intraoperative times compared to conventional navigation and metal 3D-printed surgical guides. The AR system, while exhibiting lower accuracy, exhibited efficiency, making it a promising option for HTO procedures. The preoperative planning time for the AR system was notably shorter (4 min) compared to conventional navigation (30.5 min) and metal guides (75.5 min). Intraoperative time for AR lasted 8.5 min, considerably faster than that of conventional navigation (31.5 min) and metal guides (10.5 min). CONCLUSIONS: The AR navigation system presents a transformative approach to HTO, offering a trade-off between accuracy and efficiency. Ongoing improvements, such as the incorporation of two-stage registration and pointing devices, could further enhance precision. While the system may be less accurate, its efficiency renders it a potential breakthrough in orthopedic surgery, particularly for reducing unnecessary harm and streamlining surgical procedures.

Lysophosphatidylcholine Impairs the Mitochondria Homeostasis Leading to Trophoblast Dysfunction in Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM) is a common pregnancy disorder associated with an increased risk of pre-eclampsia and macrosomia. Recent research has shown that the buildup of excess lipids within the placental trophoblast impairs mitochondrial function. However, the exact lipids that impact the placental trophoblast and the underlying mechanism remain unclear. GDM cases and healthy controls were recruited at Kaohsiung Medical University Hospital. The placenta and cord blood were taken during birth. Confocal and electron microscopy were utilized to examine the morphology of the placenta and mitochondria. We determined the lipid composition using liquid chromatography-mass spectrometry in data-independent analysis mode (LC/MSE). In vitro studies were carried out on choriocarcinoma cells (JEG3) to investigate the mechanism of trophoblast mitochondrial dysfunction. Results showed that the GDM placenta was distinguished by increased syncytial knots, chorangiosis, lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1) overexpression, and mitochondrial dysfunction. Lysophosphatidylcholine (LPC) 16:0 was significantly elevated in the cord blood LDL of GDM patients. In vitro, we demonstrated that LPC dose-dependently disrupts mitochondrial function by increasing reactive oxygen species (ROS) levels and HIF-1α signaling. In conclusion, highly elevated LPC in cord blood plays a pivotal role in GDM, contributing to trophoblast impairment and pregnancy complications.

Prognosis Prediction in Head and Neck Squamous Cell Carcinoma by Radiomics and Clinical Information.

(1) Background: head and neck squamous cell carcinoma (HNSCC) is a common cancer whose prognosis is affected by its heterogeneous nature. We aim to predict 5-year overall survival in HNSCC radiotherapy (RT) patients by integrating radiomic and clinical information in machine-learning models; (2) Methods: HNSCC radiotherapy planning computed tomography (CT) images with RT structures were obtained from The Cancer Imaging Archive. Radiomic features and clinical data were independently analyzed by five machine-learning algorithms. The results were enhanced through a voted ensembled approach. Subsequently, a probability-weighted enhanced model (PWEM) was generated by incorporating both models; (3) Results: a total of 299 cases were included in the analysis. By receiver operating characteristic (ROC) curve analysis, PWEM achieved an area under the curve (AUC) of 0.86, which outperformed both radiomic and clinical factor models. Mean decrease accuracy, mean decrease Gini, and a chi-square test identified T stage, age, and disease site as the most important clinical factors in prognosis prediction; (4) Conclusions: our radiomic-clinical combined model revealed superior performance when compared to radiomic and clinical factor models alone. Further prospective research with a larger sample size is warranted to implement the model for clinical use.

Efficacy of T-cell assays for the diagnosis of primary defects in cytotoxic lymphocyte exocytosis.

ABSTRACT: Primary hemophagocytic lymphohistiocytosis (HLH) is a life-threatening disorder associated with autosomal recessive variants in genes required for perforin-mediated lymphocyte cytotoxicity. A rapid diagnosis is crucial for successful treatment. Although defective cytotoxic T lymphocyte (CTL) function causes pathogenesis, quantification of natural killer (NK)-cell exocytosis triggered by K562 target cells currently represents a standard diagnostic procedure for primary HLH. We have prospectively evaluated different lymphocyte exocytosis assays in 213 patients referred for evaluation for suspected HLH and related hyperinflammatory syndromes. A total of 138 patients received a molecular diagnosis consistent with primary HLH. Assessment of Fc receptor-triggered NK-cell and T-cell receptor (TCR)-triggered CTL exocytosis displayed higher sensitivity and improved specificity for the diagnosis of primary HLH than routine K562 cell-based assays, with these assays combined providing a sensitivity of 100% and specificity of 98.3%. By comparison, NK-cell exocytosis after K562 target cell stimulation displayed a higher interindividual variability, in part explained by differences in NK-cell differentiation or large functional reductions after shipment. We thus recommend combined analysis of TCR-triggered CTL and Fc receptor-triggered NK-cell exocytosis for the diagnosis of patients with suspected familial HLH or atypical manifestations of congenital defects in lymphocyte exocytosis.

Fatigue following head and neck cancer radiotherapy: a systematic review of dose correlates.

INTRODUCTIONS: Radical radiotherapy (RT) is the cornerstone of Head and Neck (H&N) cancer treatment, but it often leads to fatigue due to irradiation of brain structures, impacting patient quality of life. OBJECTIVE: This study aimed to systematically investigate the dose correlates of fatigue after H&N RT in brain structures. METHODS: The systematic review included studies that examined the correlation between fatigue outcomes in H&N cancer patients undergoing RT at different time intervals and brain structures. PubMed, Scopus, and WOS databases were used in the systematic review. A methodological quality assessment of the included studies was conducted following the PRISMA guidelines. After RT, the cohort of H&N cancer patients was analyzed for dose correlations with brain structures and substructures, such as the posterior fossa, brainstem, cerebellum, pituitary gland, medulla, and basal ganglia. RESULT: Thirteen studies meeting the inclusion criteria were identified in the search. These studies evaluated the correlation between fatigue and RT dose following H&N RT. The RT dose ranged from 40 Gy to 70 Gy. Most of the studies indicated a correlation between the trajectory of fatigue and the dose effect, with higher levels of fatigue associated with increasing doses. Furthermore, five studies found that acute and late fatigue was associated with dose volume in specific brain structures, such as the brain stem, posterior fossa, cerebellum, pituitary gland, hippocampus, and basal ganglia. CONCLUSION: Fatigue in H&N RT patients is related to the radiation dose received in specific brain areas, particularly in the posterior fossa, brain stem, cerebellum, pituitary gland, medulla, and basal ganglia. Dose reduction in these areas may help alleviate fatigue. Monitoring fatigue in high-risk patients after radiation therapy could be beneficial, especially for those experiencing late fatigue.

Asthma Care from Home: Study protocol for an effectiveness-implementation evaluation of a virtually enabled asthma care initiative in children in rural NSW.

BACKGROUND: Asthma is the leading source of unscheduled hospitalisation in Australian children, with a high burden placed upon children, their parents/families, and the healthcare system. In Australia, there are widening disparities in paediatric asthma care including inequitable access to comprehensive ongoing and planned asthma care for children. METHODS: The Asthma Care from Home Project is a comprehensive virtually enabled asthma model of care that aims to a. supports families, communities and healthcare providers, b. flexible and locally acceptable, and c. allow for adoption of innovations such as digital technologies so that asthma care can be provided "from home", reduce potentially preventable asthma hospitalisation, and ensure satisfaction at a patient, family, and healthcare provider level. The model of care includes standardisation of discharge care through provision of an asthma discharge resource pack containing individual asthma action plan, follow-up letters for the child's general practitioner (GP) and school/child care, and access to online asthma educational sessions and resource; post-discharge care coordination through text message reminders for families for regular GP review, email correspondence with their child's GP and school/childcare; and virtual home visits to discuss home environmental triggers, provide personalised asthma education and respond to parental concerns relating to their child's asthma. This study is comprised of three components: 1) a quasi-experimental pre/post impact evaluation assessing the impact of the model on healthcare utilisation and asthma control measures; 2) a mixed-methods implementation evaluation to understand how and why our intervention was effective or ineffective in producing systems change; 3) an economic evaluation to assess the cost-effectiveness of the proposed model of care from a family and health services perspective. DISCUSSION: This study aims to improve access to asthma care for children in rural and remote areas. Implementation evaluation and economic evaluation will provide insights into the sustainability and scalability of the asthma model of care.

Deep Learning-Based Prediction Model for the Cobb Angle in Adolescent Idiopathic Scoliosis Patients.

Scoliosis, characterized by spine deformity, is most common in adolescent idiopathic scoliosis (AIS). Manual Cobb angle measurement limitations underscore the need for automated tools. This study employed a vertebral landmark extraction method and Feedforward Neural Network (FNN) to predict scoliosis progression in 79 AIS patients. The novel intervertebral angles matrix format showcased results. The mean absolute error for the intervertebral angle progression was 1.5 degrees, while the Pearson correlation of the predicted Cobb angles was 0.86. The accuracy in classifying Cobb angles (<15°, 15-25°, 25-35°, 35-45°, >45°) was 0.85, with 0.65 sensitivity and 0.91 specificity. The FNN demonstrated superior accuracy, sensitivity, and specificity, aiding in tailored treatments for potential scoliosis progression. Addressing FNNs' over-fitting issue through strategies like "dropout" or regularization could further enhance their performance. This study presents a promising step towards automated scoliosis diagnosis and prognosis.

Exploration of Textile-Silicone Composites and Materials for Personal Impact-Resistant Protection.

Conventional cushioning materials such as silicone sheets which have been recommended for resisting impact generally cause discomfort to the wearer from heat and perspiration. With the increasing need for personal protective equipment, textile-silicone composite structures are proposed in this study to reduce acute impact and moisture while enhancing thermal comfort. The influence of the composite structure and thickness on the mechanical and thermal properties of textile-silicone materials are systematically investigated. The results show that an additional knitted powernet fabric as a composite material can significantly improve the tensile properties of silicone rubber by up to 315%. However, only a slight improvement is found in the thermal conductivity (up to 16%), compression elasticity (up to 18%) and force reduction performance (up to 3.6%). As compared to inlaid spacer fabric, which has also been used for cushioning and preserving thermal comfort, the textile-silicone composites have higher tensile and compression elasticity, exhibit force reduction with the largest difference of 43% and are more thermally conductive, with increases more than 38%. The findings of this study introduced a cost-effective new silicone-textile composite for optimal impact protection and wear comfort for protective applications.

Risk Factors and Clinical Prognosis Associated With RSV-ALRI Intensive Care Unit Admission in Children <2 Years of Age: A Multicenter Study.

BACKGROUND: Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infections (ALRIs) in children <2 years of age. Currently, there are limited data on risk factors for very severe RSV-ALRI requiring intensive care unit (ICU) admission. METHODS: We conducted a case-control study of children <2 years old admitted with RSV-ALRI to the Sydney Children's Hospital Network, comprising 2 large tertiary pediatric hospitals. Cases were children with laboratory-confirmed RSV-ALRI admitted to ICU, and controls were (1:2, matched on date of admission) children hospitalized with RSV-ALRI but not requiring ICU transfer. Data on risk factors were retrieved from the electronic medical record system. Adjusted odds ratios (aORs) with 95% confidence intervals (95% CI) associated with risk factors for ICU admission and the association with clinical and treatment factors were determined from logistic regression models. RESULTS: A total of 44 (44%) of 100 cases and 90 (48.1%) of 187 controls were male. Age <6 months and preterm births were associated with a 2.10-fold (95% CI: 1.14-3.79) and 2.35-fold (95% CI: 1.26-4.41) increased risk in ICU admissions, respectively. The presence of any chronic health condition was a significant risk factor for ICU admission. The clinical presentations on admission more commonly seen in cases were apnea (aOR: 5.01, 95% CI: 1.50-17.13) and respiratory distress (aOR: 15.91, 95% CI: 4.52-55.97). Cases were more likely to be hospitalized for longer duration and require respiratory support. CONCLUSIONS: Our results can be translated into a clinical risk algorithm to identify children at risk of very severe RSV disease.

Progressive ataxia, ophthalmoparesis, and hypogonadotropic hypogonadism in a family with a novel variant in the KIFBP gene.

A novel homozygous variant in KIFBP was identified in a consanguineous family with four sibs affected by Goldberg-Sphrintzen Syndrome (GOSHS). We report for the first time, early-adulthood-onset progressive ataxia, opthalmoparesis, and hypogonadotropic hypogonadism in GOSHS.

Thermally activated delayed fluorescence tetradentate ligand-containing gold(III) complexes with preferential molecular orientation and their application in organic light-emitting devices.

A new class of thermally activated delayed fluorescence (TADF) pyridine-/pyrazine-containing tetradentate C^C^N^N gold(III) complexes have been designed and synthesized. Displaying photoluminescence quantum yields (PLQYs) of up to 0.77 in solid-state thin films, these complexes showed at-least a six-fold increase in the radiative decay rate constant (kr) in toluene upon increasing temperature from 210 to 360 K. Using variable-temperature (VT) ultrafast transient absorption (TA) spectroscopy, the reverse intersystem crossing (RISC) processes were directly observed and the activation parameters were determined, in line with the results of the Boltzmann two-level model fittings, in which the energy separation values between the lowest-lying singlet excited state (S1) and the lowest-lying triplet excited state (T1), ΔE(S1-T1), of these complexes were estimated to be in the range of 0.16-0.18 eV. Through strategic modification of the position of the electron-donating -tBu substituent in the cyclometalating ligand, the permanent dipole moments (PDMs) of these tetradentate gold(III) emitters could be manipulated to enhance their horizontal alignment in the emitting layer of organic light-emitting devices (OLEDs). Consequently, the resulting vacuum-deposited OLEDs demonstrated a 30% increase in the theoretical out-coupling efficiency (ηout), as well as promising electroluminescence (EL) performance with maximum external quantum efficiencies (EQEs) of up to 15.7%.

When is it safe to return to driving after distal radius fracture fixation? A prospective study.

INTRODUCTION: After surgical fixation of distal radius fractures, many patients are keen to return to driving. There are however limited guidelines assisting surgeons. The aims of this study were to determine when patients could return to driving safely after distal radius fracture fixation and determine the clinical parameters (range of motion and grip strength) that patients needed to achieve before return to safe driving could be advised. MATERIALS AND METHODS: A prospective grant-funded clinical study was conducted. Patients above the age of 21 years who underwent surgical fixation with a volar plate, possessed a class 3 standard motorcar license, and were regular drivers were recruited in a single institution from 2017 to 2019. A hand surgeon and an occupational therapist who sees routine hand therapy cases, assessed the patients at regular intervals from 2 to 12-weeks post-surgery. Clinical parameters of pain, wrist range of motion and grip strength were measured. Patients underwent off and on-road driving assessments. RESULTS: A total of 26 patients were recruited, with 21 successfully completing the driving assessment. Median time post-surgery to passing the driving test was 6 and 8-weeks for off and on-road assessments respectively. Pain score was observed to decrease over time, with a significant decrease from week 2 to week 4. Range of motion improved over time, with maximal improvement between 2 to 4-weeks post-surgery. When compared with the unaffected wrist, the difference in pronation, supination and radial deviation in the affected hand was consistently no longer statistically significant 4 to 6-weeks post-surgery. CONCLUSION: Patients with isolated surgically treated distal radius fractures can be recommended for a driving assessment as early as 4-6 weeks post-surgery if pain control is adequate, and clinical parameters for pronation and supination are met.

Effects of social buffering on fear extinction in adolescent rats.

Across social species, the presence of another individual can reduce stress reactions to adverse stimuli, a phenomenon known as social buffering. The present study investigated whether social buffering influences the expression and extinction of learned fear in adolescence, a developmental period of diminished fear inhibition and increased social interaction. Quality of maternal care and degree of social investigation were examined as factors that may influence social buffering. In adolescence, male rats were fear conditioned and then given extinction training either in the presence of a same-age rat or alone. Animals were then tested alone for extinction retention. In two experiments, the presence of a conspecific robustly reduced conditioned fear responses during extinction training. Interestingly, a persistent social buffering effect was observed when the extinction and conditioning contexts had prominent differences in features (Experiment 1), but not when these contexts were relatively similar (Experiment 2). Neither quality of maternal care nor degree of social investigation predicted the effects of social buffering. These findings suggest that social buffering robustly dampens fear responses during adolescence when a peer is present and this suppression can persist, in some instances, even when the peer is absent.

Assessing Older Adults' Functional Visual Performance Using the Performance Quality Rating Scale.

Visual impairment has distinct impacts on the activities of older adults. Quantifying the functional impact of visual loss would facilitate targeted rehabilitation. The objectives of this study were to: (1) develop an observational assessment of the functional visual performance of older adults using the Performance Quality Rating Scale (PQRS); (2) test the feasibility and inter-rater agreement in a pilot sample of older adults with visual impairment. A convenience sample of older adults with vision loss (N = 20) performed seven pre-selected activities. Performance was videoed (N = 126 videos) and rated by two raters using specific operational definitions. All participants completed the seven activities with the given resources and 90% of videos were successfully rated using the developed PQRS. Inter-rater agreement was substantial (weighted Kappa = 0.71; 95% confidence interval [CI] = [0.64, 0.79]) for all activities. The developed PQRS for functional vision is feasible, with substantial inter-rater agreement, to assess functional vision of older adults in an outpatient setting.

Assessing older adults' use of vision using the Performance Quality Rating Scale.Visual impairment has different impacts on the everyday activities of older adults. Assessing the specific impact would help therapists to provide rehabilitation targeting their daily challenges. The objectives of this study were (1) to develop an assessment of how older adults use their vision using the Performance Quality Rating Scale (PQRS); (2) to test the possibility of using, and the agreement of using this tool between two raters in a pilot sample. Twenty older adults with vision loss performed seven activities in an outpatient clinic. In total, 126 videos of their performances were rated by two raters using the PQRS. 90% of the videos were successfully rated using the developed PQRS with good agreement between the raters. The developed PQRS can possibly be used to assess how older adults use their vision for daily activities in an outpatient setting.

A Scientometric Analysis and Visualization of Prosthetic Foot Research Work: 2000 to 2022.

This study aims to highlight recent research work on topics around prosthetic feet through a scientometric analysis and historical review. The most cited publications from the Clarivate Analytics Web of Science Core Collection database were identified and analyzed from 1 January 2000 to 31 October 2022. Original articles, reviews with full manuscripts, conference proceedings, early access documents, and meeting abstracts were included. A scientometric visualization analysis of the bibliometric information related to the publications, including the countries, institutions, journals, references, and keywords, was conducted. A total of 1827 publications met the search criteria in this study. The related publications grouped by year show an overall trend of increase during the two decades from 2000 to 2022. The United States is ranked first in terms of overall influence in this field (n = 774). The Northwestern University has published the most papers on prosthetic feet (n = 84). Prosthetics and Orthotics International has published the largest number of studies on prosthetic feet (n = 151). During recent years, a number of studies with citation bursts and burst keywords (e.g., diabetes, gait, pain, and sensor) have provided clues on the hotspots of prosthetic feet and prosthetic foot trends. The findings of this study are based on a comprehensive analysis of the literature and highlight the research topics on prosthetic feet that have been primarily explored. The data provide guidance to clinicians and researchers to further studies in this field.

Social and ecological disparities in anaemia among adolescent girls 15-19 years old in Nepal.

OBJECTIVE: Adolescent girls are at risk of anaemia due to increased nutrient demands because of growth, menstrual blood loss and possible pregnancies. Sociocultural and household conditions influence their anaemia risk. We aimed to identify the sociocultural and economic factors associated with anaemia among adolescent girls in Nepal. DESIGN: The Nepal Demographic and Health Surveys (NDHS) conducted in 2006, 2011 and 2016 were pooled for secondary analysis. We used data on haemoglobin measurements for anaemia and conducted bivariate and multivariable logistic regression analyses to identify factors associated with anaemia. SETTING: Nationally representative NDHS households with adolescent girls 15-19 years of age. PARTICIPANTS: Non-pregnant adolescent girls 15-19 years, with a haemoglobin measurement (n = 3731). RESULTS: The overall prevalence of anaemia among adolescent girls was 39·6 %. Adolescents from socially disadvantaged caste/ethnicity groups were 1·42 times (95 % CI: 1·13, 1·78) more likely to have anaemia compared with those from Brahmin/Chhetri households. We found a counter-intuitive association between socio-economic status and anaemia where adolescents from the middle (adjusted OR (aOR) 1·37, 95 % CI: 1·01, 1·85) and highest (aOR 1·74, 95 % CI: 1·18, 2·56) quintiles were at increased odds of anaemia. Relative geographical inequality was observed where adolescents from the Terai region had 3·5 times (95 % CI: 2·32, 5·33) higher odds of anaemia. CONCLUSIONS: The disparities in the distribution of anaemia among adolescents by caste/ethnicity groups, wealth quintiles and geographical regions are evident. Reducing the anaemia burden will require addressing the social determinants of anaemia by allocating resources and expanding anaemia prevention programmes to target adolescents at higher risk.