Variations of Major Glucosinolates in Diverse Chinese Cabbage (Brassica rapa ssp. pekinensis) Germplasm as Analyzed by UPLC-ESI-MS/MS.

In this study, the variability of major glucosinolates in the leaf lamina of 134 Chinese cabbage accessions was investigated using Acquity ultra-performance liquid chromatography (UPLC-ESI-MS/MS). A total of twenty glucosinolates were profiled, of which glucobrassicanapin and gluconapin were identified as the predominant glucosinolates within the germplasm. These two glucosinolates had mean concentration levels above 1000.00 µmol/kg DW. Based on the principal component analysis, accessions IT186728, IT120044, IT221789, IT100417, IT278620, IT221754, and IT344740 were separated from the rest in the score plot. These accessions exhibited a higher content of total glucosinolates. Based on the VIP values, 13 compounds were identified as the most influential and responsible for variation in the germplasm. Sinigrin (r = 0.73), gluconapin (r = 0.78), glucobrassicanapin (r = 0.70), epiprogoitrin (r = 0.73), progoitrin (r = 0.74), and gluconasturtiin (r = 0.67) all exhibited a strong positive correlation with total glucosinolate at p < 0.001. This indicates that each of these compounds had a significant influence on the overall glucosinolate content of the various accessions. This study contributes valuable insights into the metabolic diversity of glucosinolates in Chinese cabbage, providing potential for breeding varieties tailored to consumer preferences and nutritional demands.

The relationship between event boundary strength and pattern shifts across the cortical hierarchy during naturalistic movie-viewing.

Our continuous experience is spontaneously segmented by the brain into discrete events. However, the beginning of a new event (an event boundary) is not always sharply identifiable: phenomenologically, event boundaries vary in salience. How are the response profiles of cortical areas at event boundaries modulated by boundary strength during complex, naturalistic movie-viewing? Do cortical responses scale in a graded manner with boundary strength, or do they merely detect boundaries in a binary fashion? We measured "cortical boundary shifts" as transient changes in multi-voxel patterns at event boundaries with different strengths (weak, moderate, and strong), determined by across-subject agreement. Cortical regions with different processing timescales were examined. In auditory areas, which have short timescales, cortical boundary shifts exhibited a clearly graded profile both in group-level and individual-level analyses. In cortical areas with long timescales, including the default mode network, boundary strength modulated pattern shift magnitude at the individual subject level. We also observed a positive relationship between boundary strength and the extent of temporal alignment of boundary shifts across different levels of the cortical hierarchy. A strictly nested bottom-up hierarchical structure was not necessary to observe this relationship. Additionally, hippocampal activity was highest at event boundaries for which cortical boundary shifts were most aligned across hierarchical levels. Overall, we found that event boundary strength modulated cortical pattern shifts strongly in sensory areas and more weakly in higher-level areas, and that stronger boundaries were associated with greater alignment of these shifts across the cortical hierarchy.

Challenges and potential improvements in the Accreditation Standards of the Korean Institute of Medical Education and Evaluation 2019 (ASK2019) derived through meta-evaluation: a cross-sectional study

PURPOSE: This study aimed to identify challenges and potential improvements in Korea's medical education accreditation process according to the Accreditation Standards of the Korean Institute of Medical Education and Evaluation 2019 (ASK2019). Meta-evaluation was conducted to survey the experiences and perceptions of stakeholders, including self-assessment committee members, site visit committee members, administrative staff, and medical school professors. METHODS: A cross-sectional study was conducted using surveys sent to 40 medical schools. The 332 participants included self-assessment committee members, site visit team members, administrative staff, and medical school professors. The t-test, one-way analysis of variance and the chi-square test were used to analyze and compare opinions on medical education accreditation between the categories of participants. RESULTS: Site visit committee members placed greater importance on the necessity of accreditation than faculty members. A shared positive view on accreditation's role in improving educational quality was seen among self-evaluation committee members and professors. Administrative staff highly regarded the Korean Institute of Medical Education and Evaluation's reliability and objectivity, unlike the self-evaluation committee members. Site visit committee members positively perceived the clarity of accreditation standards, differing from self-assessment committee members. Administrative staff were most optimistic about implementing standards. However, the accreditation process encountered challenges, especially in duplicating content and preparing self-evaluation reports. Finally, perceptions regarding the accuracy of final site visit reports varied significantly between the self-evaluation committee members and the site visit committee members. CONCLUSION: This study revealed diverse views on medical education accreditation, highlighting the need for improved communication, expectation alignment, and stakeholder collaboration to refine the accreditation process and quality.

Tandem Mass Tag LC-MS/MS of Aqueous Humor From Individuals With Type 2 Diabetes Without Retinopathy Reveals Early Dysregulation of Synaptic Proteins.

Purpose: An early neurodegenerative component of diabetic retinal disease (DRD) that precedes the vascular findings of clinically diagnosed diabetic retinopathy (DR) is increasingly being recognized. However, the relevant molecular mechanisms and biomarkers for early DRD are poorly defined. The purpose of this study was to uncover novel potential mediators of early diabetic retinal neuronal dysfunction through analysis of the aqueous fluid proteome in preclinical DR. Methods: Aqueous fluid was collected from subjects with type 2 diabetes mellitus (DM) but no clinical DR and from nondiabetic controls undergoing routine cataract surgery. Preoperative spectral-domain optical coherence tomography of the macula was obtained. Tandem mass tag LC-MS/MS was performed to identify proteins differentially present in diabetic and control aqueous fluid, and proteins with >50% change and P < 0.05 were considered significant. Selected results were validated with western blot of human aqueous fluid samples. Results: We identified decreased levels of proteins implicated in neuronal synapse formation and increased levels of inflammatory proteins in the aqueous fluid from patients with type 2 DM but no DR compared with controls. Of the differentially present synaptic proteins that we identified and confirmed with western blot, the majority have not previously been linked with DRD. Conclusions: The proteomic profile of aqueous fluid from individuals with type 2 DM but no DR suggests that retinal neuronal dysfunction and inflammation represent very early events in the pathophysiology of DRD. These findings support the concept that diabetic retinal neurodegeneration precedes vascular pathology and reveal novel potential mediators and/or biomarkers warranting further investigation.

Beyond the surface: unraveling global health curriculum insights through interviews of learners and educators using the CIPP model.

PURPOSE: Our study aimed to delve beyond a surface-level understanding and explore the various dimensions of the global health curriculum from the perspective of both learners and educators using the Context, Input, Process, and Product (CIPP) model. METHODS: From 2020 to 2021, interviews were conducted with a total of 10 individuals, including five students who had taken at least one elective course and at least one elective research course, three teaching assistants (TA), and two faculty members who had taken more than four global health courses in multiple phases in the global health curriculum. Semi-structured interview questions based on the CIPP model were used and qualitative data were analyzed through content analysis. RESULTS: The study identified 12 sub-themes. Students held idealized views of global health careers and sought to bridge the gap through global health classes. They desired early exposure to global health courses, emphasizing both pre-medical and clinical phases. Challenges in adjusting course difficulty and recruiting faculty were identified, along with a preference for interactive teaching methods and offline discussions. The curriculum promoted reflection on medicine's essence, expanded career perspectives, and emphasized competencies like altruism, communication skills, and crisis management in the evolving global health landscape. CONCLUSION: This study showed that a comprehensive approach is possible from the perspective of learners and educators by identifying strengths, weaknesses, and the value of the curriculum's goals, plans, implementations, and results through the CIPP model. For optimal curriculum design, a sequential approach from basic to advanced courses is essential, promoting hands-on global health experiences for students.

Comparative Study on the Immunogenicity of COVID-19 mRNA Vaccines in Patients Receiving Adjuvant and Palliative Chemotherapy.

This study was conducted to investigate potential differences in vaccine efficacy between patients undergoing palliative chemotherapy and receiving adjuvant chemotherapy. Additionally, the study proved the influence of vaccination timing on vaccine efficacy during active chemotherapy. Anti-receptor-binding domain (RBD) IgG binding antibody assays and surrogate neutralizing antibody assays were performed after BNT162b2 or mRNA-1273 vaccination in 45 solid cancer patients (23 adjuvant and 22 palliative chemotherapy) and in 24 healthy controls before vaccination (baseline), at every two to four weeks after the first (post-dose 1) and the second vaccination (post-dose 2). The levels of anti-RBD IgG and neutralizing antibodies increased significantly from baseline through post-dose 1 to post-dose 2 in all three groups. At the post-dose 1, the anti-RBD IgG and neutralizing antibody levels were significantly lower in cancer patients than in healthy controls. However, by post-dose 2, the seropositivity of anti-RBD IgG and neutralizing antibodies uniformly reached 100% across all groups, with no significant disparity in antibody levels among the three groups. Moreover, the antibody titers were not significantly different between patients with a vaccine and chemotherapy interval of more than 14 days or those with less than 14 days. This study demonstrated that after second doses of mRNA COVID-19 vaccines, humoral immune responses in patients receiving chemotherapy were comparable to those of healthy controls, regardless of whether the purpose of the anti-cancer treatment was palliative or adjuvant. Furthermore, the timing of vaccination did not affect the level of humoral immunity after the second vaccination.

Comprehensive evaluation of nutritional components, bioactive metabolites, and antioxidant activities in diverse sorghum (Sorghum bicolor (L.) Moench) landraces.

Sorghum, one of the prospective crops for addressing future food and nutrition security, has received attention in recent years due to its health-promoting compounds. It is known that several environmental and genetic factors affect the metabolite contents of dietary crops. This study investigated the diversity of different nutrients, functional metabolites, and antioxidant activity using three different assays in 53 sorghum landraces from Korea, China, Japan, Ethiopia, and South Africa. The effects of origin and seed color variations were also investigated. Total phenolic (TPC), total tannin (TTC), total fat, total protein, total dietary fiber, and total crude fiber contents all varied significantly among the sorghum landraces (p < 0.05). Using a gas chromatography-flame ionization detector, palmitic, stearic, oleic, linoleic, and linolenic acids were detected in all the sorghum landraces, and their content significantly varied (p < 0.05). Furthermore, four 3-deoxyanthocyanidins (luteolinidin, apigeninidin, 5-methoxyluteolinidin, and 7-methoxyapigeninidin) and two flavonoids (luteolin and apigenin) were detected in most of the landraces using liquid chromatography-tandem mass spectrometry, and their concentrations also significantly varied. Statistical analyses supported by multivariate tools demonstrated that seed color variation had a significant effect on TPC, TTC, DPPH• and ABTS•+ scavenging activities, and ferric-reducing antioxidant power, with yellow landraces having the highest and white landraces having the lowest values. Seed color variation also had a significant effect on dietary fiber, linoleic acid, linolenic acid, and luteolin contents. In contrast, all nutritional components and fatty acids except total protein and oleic acid were significantly affected by origin, while most 3-deoxyanthocyanidins and flavonoids were unaffected by both origin and seed color differences. This is the first study to report the effect of origin on sorghum seed metabolites and antioxidant activities, laying the groundwork for future studies. Moreover, this study identified superior landraces that could be good sources of health-promoting metabolites.

Effects of SKCPT on Osteoarthritis in Beagle Meniscectomy and Cranial Cruciate Ligament Transection Models.

Osteoarthritis (OA) affects >500 million people globally, and this number is expected to increase. OA management primarily focuses on symptom alleviation, using non-steroidal anti-inflammatory drugs, including Celecoxib. However, such medication has serious side effects, emphasizing the need for disease-specific treatment. The meniscectomy and cranial cruciate ligament transection (CCLx)-treated beagle dog was used to investigate the efficacy of a modified-release formulation of SKI306X (SKCPT) from Clematis mandshurica, Prunella vulgaris, and Trichosanthes kirilowii in managing arthritis. SKCPT's anti-inflammatory and analgesic properties have been assessed via stifle circumference, gait, incapacitance, histopathology, and ELISA tests. The different SKCPT concentrations and formulations also affected the outcome. SKCPT improved the gait, histopathological, and ELISA OA assessment parameters compared to the control group. Pro-inflammatory cytokines and matrix metalloproteinases were significantly lower in the SKCPT-treated groups than in the control group. This study found that SKCPT reduces arthritic lesions and improves abnormal gait. The 300 mg modified-release formulation was more efficacious than others, suggesting a promising approach for managing OA symptoms and addressing disease pathogenesis. A high active ingredient level and a release pattern make this formulation effective for twice-daily arthritis treatment.

Word-timestamped transcripts of two spoken narrative recall functional neuroimaging datasets.

After watching audiovisual movies, human participants produced spoken narrative recollections during functional magnetic resonance imaging (fMRI); presented here are word-level timestamps of their speech, temporally aligned to the publicly shared fMRI data. For the "FilmFestival" dataset, twenty participants watched ten short audiovisual movies, approximately 2-8 minutes each. For the "Sherlock" dataset, seventeen participants watched the first half of the first episode of BBC's Sherlock (48 minutes). After viewing, participants then verbally described what they remembered about the movies in their own words. Participants' speech was recorded using an MR-compatible microphone. The audio recordings were transcribed, then timestamped by a forced aligner; missing timestamps were filled in manually by human transcriptionists referencing the audio recording. Each file contains the participant's recall word by word, onset of each word in seconds with 1/10th-second precision, and the corresponding fMRI volume number (TR). This dataset can be used to investigate topics such as naturalistic memory and language production.

Exploring the potentials of sorghum genotypes: a comprehensive study on nutritional qualities, functional metabolites, and antioxidant capacities.

Introduction: Sorghum, long regarded as one of the most underutilized crops, has received attention in recent years. As a result, conducting multidisciplinary studies on the potential and health benefits of sorghum resources is vital if they are to be fully exploited. In this study, the nutritional contents, functional metabolites, and antioxidant capacities of 23 sorghum breeding lines and three popular cultivars were assessed. Materials and method: All of the sorghum genotypes were grown under the same conditions, and mature seeds were hand-harvested. The metabolite contents and antioxidant capacities of sorghum seeds were assessed using standard protocols. Fatty acids were quantified using a gas chromatography-flame ionization detector, whereas flavonoids and 3-deoxyanthocyanidins were analyzed using a liquid chromatography-tandem mass spectrometry method. The data were analyzed using both univariate and multivariate statistical approaches. Results and discussion: Total protein (9.05-14.61%), total fat (2.99-6.91%), crude fiber (0.71-2.62%), dietary fiber (6.72-16.27%), total phenolic (0.92-10.38 mg GAE/g), and total tannin (0.68-434.22 mg CE/g) contents varied significantly across the sorghum genotypes (p < 0.05). Antioxidant capacity, measured using three assays, also differed significantly. Five fatty acids, including palmitic, stearic, oleic, linoleic, and linolenic acids, were found in all the sorghum genotypes with statistically different contents (p < 0.05). Furthermore, the majority of the sorghum genotypes contained four 3-deoxyanthocyanidins, including luteolinidin, apigeninidin, 5-methoxyluteolinidin, and 7-methoxyapigeninidin, as well as two dominant flavonoids, luteolin and apigenin. Compared to the cultivars, some breeding lines had significantly high levels of metabolites and antioxidant activities. On the other hand, statistical analysis showed that total tannin, total phenolic, and antioxidant capacities varied significantly across white, yellow, and orange genotypes. Principal component analysis was used to differentiate the sorghum genotypes based on seed color and antioxidant index levels. Pearson's correlation analysis revealed strong links between biosynthetically related metabolites and those with synergistic antioxidant properties. Conclusion: This research demonstrated the diversity of the sorghum resources investigated. Those genotypes with high levels of nutritional components, functional metabolites, and antioxidant activities could be used for consumption and breeding programs.

Genome-Wide Association Study of Seed Morphology Traits in Senegalese Sorghum Cultivars.

Sorghum is considered the fifth most important crop in the world. Despite the potential value of Senegalese germplasm for various traits, such as resistance to fungal diseases, there is limited information on the study of sorghum seed morphology. In this study, 162 Senegalese germplasms were evaluated for seed area size, length, width, length-to-width ratio, perimeter, circularity, the distance between the intersection of length & width (IS) and center of gravity (CG), and seed darkness and brightness by scanning and analyzing morphology-related traits with SmartGrain software at the USDA-ARS Plant Science Research Unit. Correlations between seed morphology-related traits and traits associated with anthracnose and head smut resistance were analyzed. Lastly, genome-wide association studies were performed on phenotypic data collected from over 16,000 seeds and 193,727 publicly available single nucleotide polymorphisms (SNPs). Several significant SNPs were found and mapped to the reference sorghum genome to uncover multiple candidate genes potentially associated with seed morphology. The results indicate clear correlations among seed morphology-related traits and potential associations between seed morphology and the defense response of sorghum. GWAS analysis listed candidate genes associated with seed morphologies that can be used for sorghum breeding in the future.

Three-dimensional soft tissue landmark detection with marching cube algorithm.

Current method of analyzing three-dimensional soft tissue data, especially in the frontal view, is subjective and has poor reliability. To overcome this limitation, the present study aimed to introduce a new method of analyzing soft tissue data reconstructed by marching cube algorithm (Program S) and compare it with a commercially available program (Program A). Cone-beam computed tomography images of 42 patients were included. Two orthodontists digitized six landmarks (pronasale, columella, upper and lower lip, right and left cheek) twice using both programs in two-week intervals, and the reliability was compared. Furthermore, computer-calculated point (CC point) was developed to evaluate whether human error could be reduced. The results showed that the intra- and inter-examiner reliability of Program S (99.7-100% and 99.9-100%, respectively) were higher than that of Program A (64.0-99.9% and 76.1-99.9%, respectively). Moreover, the inter-examiner difference of coordinate values and distances for all six landmarks in Program S was lower than Program A. Lastly, CC point was provided as a consistent single point. Therefore, it was validated that this new methodology can increase the intra- and inter-examiner reliability of soft tissue landmark digitation and CC point can be used as a landmark to reduce human error.

Nosocomial Outbreak of COVID-19 from a Kidney Transplant Patient: Necessity of a Longer Isolation Period in Immunocompromised Patients.

BACKGROUND: Determination of the release from isolation for coronavirus disease 2019 (COVID-19) in immunocompromised patients who need additional hospitalization for treatment of non-COVID-19 related disease is important to prevent nosocomial transmission. However, there is insufficient evidence for an extended isolation period. MATERIALS AND METHODS: In September 2021, when the Delta variant was dominant, a nosocomial outbreak of COVID-19 occurred in the nephrology ward of a tertiary hospital in Gwangju, Korea. We conducted epidemiological investigations and whole-genome sequencing (WGS) of this virus. RESULTS: A man who underwent kidney transplantation was admitted to our hospital for the treatment of acute kidney injury. He was diagnosed with asymptomatic COVID-19 infection during a pre-admission screening test on September 1, 2021 and underwent isolation. After 10 days of isolation in the COVID-19-designated ward, he was transferred to the general nephrology ward. He underwent steroid pulse therapy (September 17 to September 23, >60 mg/day prednisolone) due to acute T-cell rejection. On September 28, 2021, the first patient with COVID-19 was identified in the nephrology ward, and a rapid-response team was activated to identify additional patients with COVID-19 and prevent the spread of COVID-19. Epidemiological investigations revealed that 12 patients, two caregivers, and three healthcare workers from the nephrology ward were diagnosed with COVID-19. The WGS of specimens from 14 nosocomial outbreak samples and released an index patient exhibited the same Delta variant originating from the B.1.617.2 lineage. This hospital-acquired COVID-19 outbreak in the nephrology ward resulted in two (11.7%) deaths in patients who underwent kidney transplantation. CONCLUSION: We demonstrated that an immunocompromised patient can cause a nosocomial outbreak due to the prolonged shedding of infectious viruses. Prolonged isolation in patients under active immunosuppressive therapy may be necessary to prevent transmission, especially in the hospital setting.

Variability of Glucosinolates in Pak Choy (Brassica rapa subsp. chinensis) Germplasm.

Glucosinolates are sulfur-containing phytochemicals generally abundant in cruciferous vegetables such as pak choy. Glucosinolates participate in a range of biological activities essential for promoting a healthy human body. In this study, we aimed to elucidate glucosinolate variability present in pak choy germplasm that are under conservation at the Rural Development Administration Genebank, Jeonju, Republic of Korea. The Acquity Ultra-Performance Liquid Chromatography (UHPLC) analytical system was used in profiling the glucosinolate content in leaf samples of various accessions. We identified a total of 17 glucosinolates in the germplasm. Based on principal compoment analysis performed, three separate groups of the accessions were obtained. Group 1 contained the cultivar cheongsacholong which recorded high content of glucobrassicin (an indole), glucoerucin (aliphatic), gluconasturtiin (aromatic) and glucoberteroin (aliphatic). Group 2 consisted of six accessions, BRA77/72, Lu ling gaogengbai, 9041, Wuyueman, RP-75 and DH-10, predominatly high in aliphatic compounds including glucoiberin, glucocheirolin, and sinigrin. Group 3 comprised the majority of the accessions which were characterized by high content of glucoraphanin, epiprogoitrin, progoitrin, and glucotropaeolin. These results revealed the presence of variability among the pak choy germplasm based on their glucosinolate content, providing an excellent opprtunity for future breeding for improved glucosinolate content in the crop.

Genome-Wide Association Study for Agro-Morphological Traits in Eggplant Core Collection.

Eggplant is one of the most economically and nutritionally important vegetables worldwide. The study of the association of phenotypic traits with genetic factors is vital for the rapid and efficient identification and selection of eggplant genetic resources for breeding purposes with desired traits. The eggplant resources (587) collected from different countries, including Korea, were used for establishing the core collection. A total of 288 accessions were selected from 587 Solanum accessions based on 52 single nucleotide polymorphisms (SNPs) markers together with 17 morphological traits. This core collection was further used to analyze the genetic associations of eggplant morphological variations. A large variation was found among the evaluated eggplant accessions for some agro-morphological traits. Stem prickles and leaf prickles showed a significant positive correlation (r = 0.83***), followed by days to flowering and days to maturity (r = 0.64***). A total of 114,981 SNPs were filtered and used for phylogenetic tree analysis, population structure analysis, and genome-wide association study (GWAS). Among the agro-morphological traits, significantly associated SNPs were found for six traits. A total of 377 significantly associated SNPs with six agro-morphological traits were identified. These six traits and the number of SNPs were: days to maturity (51), flower size (121), fruit width (20), harvest fruit color (42), leaf prickles (38), and stem prickles (105). The largest fraction of significant SNPs (11.94%) was obtained on chromosome Ch01, followed by Ch07 and Ch06 with 11.67% and 10.08%, respectively. This study will help to develop markers linked to the most important agro-morphological traits of eggplant genetic resources and support the selection of desirable traits for eggplant breeding programs.

Mutations in OsDET1, OsCOP10, and OsDDB1 confer embryonic lethality and alter flavonoid accumulation in Rice (Oryza sativa L.) seed.

Morphological and biochemical changes accompanying embryogenesis and seed development are crucial for plant survival and crop productivity. Here, we identified a novel yellowish-pericarp embryo lethal (yel) mutant of the japonica rice cultivar Sindongjin (Oryza sativa L.), namely, yel-sdj. Seeds of the yel-sdj mutant showed a yellowish pericarp and black embryo, and were embryonic lethal. Compared with wild-type seeds, the yel-sdj mutant seeds exhibited significantly reduced grain size, grain weight, and embryo weight, and a remarkably lower rate of embryo retention in kernels subjected to milling. However, the volume of air space between embryo and endosperm, density of embryo, and total phenolic content (TPC) and antioxidant activity of mature grains were significantly higher in the yel-sdj mutant than in the wild type. Genetic analysis and mapping revealed that the yel-sdj mutant was non-allelic to the oscop1 null mutants yel-hc, yel-cc, and yel-sk, and its phenotype was controlled by a single recessive gene, LOC_Os01g01484, an ortholog of Arabidopsis thaliana DE-ETIOLATED 1 (DET1). The yel-sdj mutant carried a 7 bp deletion in the second exon of OsDET1. Seeds of the osdet1 knockout mutant, generated via CRISPR/Cas9-based gene editing, displayed the yel mutant phenotype. Consistent with the fact that OsDET1 interacts with CONSTITUTIVE PHOTOMORPHOGENIC 10 (OsCOP10) and UV-DAMAGED DNA BINDING PROTEIN 1 (OsDDB1) to form the COP10-DET1-DDB1 (CDD), seeds of oscop10 and osddb1 knockout mutants also showed the yel phenotype. These findings will enhance our understanding of the functional roles of OsDET1 and the CDD complex in embryogenesis and flavonoid biosynthesis in rice seeds.

Meta-Selective C-H Functionalization of Arylsilanes Using a Silicon Tethered Directing Group.

We describe meta-selective C-H functionalization of arylsilanes using a Si-tethered directing group. The current method enables a selective alkenylation of arenes bearing a variety of functional groups, and several electron-deficient olefins are also applicable as coupling partners. Further functional group transformations of the silicon-tethered directing group provide multisubstituted arenes efficiently.