Mendelian randomization study of urolithiasis: exploration of risk factors using human blood metabolites.

BACKGROUND: Urolithiasis is a highly prevalent global disease closely associated with metabolic factors; however, the causal relationship between blood metabolites and urolithiasis remains poorly understood. METHOD: In our study, we employed a bi-directional two-sample Mendelian randomization (MR) analysis to investigate the causal associations between urolithiasis and metabolites. The random-effects inverse-variance weighted (IVW) estimation method was utilized as the primary approach, complemented by several other estimators including MR-Egger, weighted median, colocalization and MR-PRESSO. Furthermore, the study included replication and meta-analysis. Finally, we conducted metabolic pathway analysis to elucidate potential metabolic pathways. RESULTS: After conducting multiple tests for correction, glycerol might contribute to the urolithiasis and dehydroisoandrosterone sulfate (DHEA-S) might inhibit this process. Furthermore, several blood metabolites had shown potential associations with a causal relationship. Among the protective metabolites were lipids (dehydroisoandrosterone sulfate and 1-stearoylglycerol (1-monostearin)), amino acids (isobutyrylcarnitine and 2-aminobutyrate), a keto acid (acetoacetate) and a carbohydrate (mannose). The risk metabolites included lipids (1-palmitoylglycerophosphoethanolamine, glycerol and cortisone), a carbohydrate (erythronate), a peptide (pro-hydroxy-pro) and a fatty acid (eicosenoate). In reverse MR analysis, urolithiasis demonstrated a statistically significant causal relationship with butyrylcarnitine, 3-methyl-2-oxobutyrate, scyllo-inositol, leucylleucine and leucylalanine. However, it was worth noting that none of the blood metabolites exhibited statistical significance after multiple corrections. Additionally, we identified one metabolic pathway associated with urolithiasis. CONCLUSION: The results we obtained demonstrate the causal relevance between two metabolites and urolithiasis, as well as identify one metabolic pathway potentially associated with its development. Given the high prevalence of urolithiasis, further investigations are encouraged to elucidate the mechanisms of these metabolites and explore novel therapeutic strategies.

USP39 Promotes the Viability and Migration of Head and Neck Squamous Cell Carcinoma Cell by Regulating STAT1.

Objective: Ubiquitin-specific peptidase 39 (USP39) plays a carcinogenic role in many cancers, but little research has been conducted examining whether it is involved in head and neck squamous cell carcinoma (HNSCC). Therefore, this study explored the functional role of USP39 in HNSCC. Method: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify differentially expressed proteins (DEPs) between the HNSCC tumor and adjacent healthy tissues. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to assess the functional enrichment of DEPs. Immunohistochemistry was used to detect protein expression. The viability and migration of two HNSCC cell lines, namely CAL27 and SCC25, were detected using the cell counting kit-8 assay and a wound healing assay, respectively. Quantitative real-time PCR was used to detect the expression level of signal transducer and activator of transcription 1 (STAT1) mRNA. Results: LC-MS/MS results identified 590 DEPs between HNSCC and adjacent tissues collected from 4 patients. Through GO and KEGG pathway analyses, 34 different proteins were found to be enriched in the spliceosome pathway. The expression levels of USP39 and STAT1 were significantly higher in HNSCC tumor tissue than in adjacent healthy tissue as assessed by LC-MS/MS analysis, and the increased expression of USP39 and STAT1 protein was confirmed by immunohistochemistry in clinical samples collected from 7 additional patients with HNSCC. Knockdown of USP39 or STAT1 inhibited the viability and migration of CAL27 and SCC25 cells. In addition, USP39 knockdown inhibited the expression of STAT1 mRNA in these cells. Conclusion: Our findings indicated that USP39 knockdown may inhibit HNSCC viability and migration by suppressing STAT1 expression. The results of this study suggest that USP39 may be a potential new target for HNSCC clinical therapy or a new biomarker for HNSCC.

Pregnancy-Specific Beta-1-Glycoprotein 1 Increases HTR-8/SVneo Cell Migration through the Orai1/Akt Signaling Pathway.

The impaired invasion ability of trophoblast cells is related to the occurrence of preeclampsia (PE). We previously found that pregnancy-specific beta-1-glycoprotein 1 (PSG1) levels were decreased in the serum of individuals with early-onset preeclampsia (EOPE). This study investigated the effect of PSG1 on Orai1-mediated store-operated calcium entry (SOCE) and the Akt signaling pathway in human trophoblast cell migration. An enzyme-linked immunosorbent assay (ELISA) was used to determine the level of PSG1 in the serum of pregnant women with EOPE. The effects of PSG1 on trophoblast proliferation and migration were examined using cell counting kit-8 (CCK8) and wound healing experiments, respectively. The expression levels of Orai1, Akt, and phosphorylated Akt (p-Akt) were determined through Western blotting. The results confirmed that the serum PSG1 levels were lower in EOPE women than in healthy pregnant women. The PSG1 treatment upregulated the protein expression of Orai1 and p-Akt. The selective inhibitor of Orai1 (MRS1845) weakened the migration-promoting effect mediated by PSG1 via suppressing the Akt signaling pathway. Our findings revealed one of the mechanisms possibly involved in EOPE pathophysiology, which was that downregulated PSG1 may reduce the Orai1/Akt signaling pathway, thereby inhibiting trophoblast migration. PSG1 may serve as a potential target for the treatment and diagnosis of EOPE.

[Advances in nanobody screening technology].

Nanobody (Nb) is a novel type of antibody discovered in the serum of Camelidae. It is characterized by its small size, high specificity, stability, and ease of preparation. Nanobodies exhibit the ability to identify hidden epitopes and have diverse applications across various fields. This review aims to introduce three key stages in the screening and optimization of nanobodies, including nanobody library construction, in vitro surface display, and affinity maturation. We provide a brief description of preparation and characteristics of natural, immunological, and semi-synthetic/synthetic libraries. Additionally, we systematically explain eight in vitro display methods, including phage display, yeast display, bacterial display, ribosome display/mRNA display, and eukaryotic cell display. Furthermore, we discuss the application of yeast two-hybrid system high-throughput sequencing and mass spectrometry identification. A thorough analysis of their advantages and limitations is presented in this protocols. Finally, we summarize the platforms for in vitro or computer-aided affinity maturation techniques aimed at enhancing the functional stability of nanobodies. Consequently, this review provides a comprehensive approach to the integrated utilization of various technologies for the rapid development of stable, reliable, and specific nanobody-based drugs or diagnostic agents.

The mediating effects of motivation on the relations between occupational stress and physical activity among underresourced afterschool program staff.

OBJECTIVE: Guided by Self-Determination Theory, this study aimed to examine the potential mediating effects of autonomous and controlled motivations on physical activity (PA) experiences of afterschool program (ASP) staff with occupational stress. METHOD: A total of 58 ASP staff provided full data. Staff occupational stress and self-determination motivations for PA were assessed. Participants' daily moderate-to-vigorous physical activity (MVPA) was measured using accelerometer wear. A path analysis was used to address the research purpose. RESULTS: Occupational stress negatively and indirectly predicted daily MVPA which was mediated by controlled motivation (ß = - 4.15, p <.05). Autonomous motivation directly and positively predicted daily MVPA across all types and levels of ASP staff occupational stress (ß = 9.93, p =.01). CONCLUSIONS: Autonomous motivation is a powerful predictor of staff PA levels despite the degree to which they experience stress. In contrast, controlled motivations are more vulnerable to occupational stress, and can lead to lower MVPA. TRIAL REGISTRATION: Connect Through PLAY: A Staff-based Physical Activity Intervention for Middle School Youth (Connect). https://clinicaltrials.gov/ct2/show/NCT03732144 . Registered 11/06/2018. REGISTRATION NUMBER: NCT03732144.

Changing trajectory of daily physical activity levels among at-risk adolescents: influences of motivational mechanisms.

BACKGROUND: Guided by Self-Determination Theory (SDT), the purpose of this study was to determine changes in the 16-week moderate-to-vigorous physical activity (MVPA) trajectory of underserved adolescents who participated in the Connect through PLAY afterschool program intervention and the effects of changes in participating adolescents' intrinsic and autonomous extrinsic motivations on their MVPA trajectory over the 16-week intervention. METHODS: A subsample of 113 adolescents (56.64% female; 61.06% African American; average age = 11.29) provided complete data throughout the 16-week intervention were examined. Adolescents' objective daily MVPA was measured using 7- day accelerometer data. Changes in adolescents' intrinsic motivation and autonomous extrinsic motivation were assessed using subscales from the Intrinsic Motivation Inventory [1] and the Treatment Self-Regulation Questionnaire [2] respectively. A hierarchical linear model was built and tested to address the research aims. RESULTS: The results of hierarchical linear models showed that, on average, youth daily MVPA increased 6.36 minutes in each 8-week period. Intrinsic motivation change, but not autonomous extrinsic motivation, was a positive and significant level-2 predictor of daily MVPA changes. CONCLUSION: The findings provide significant evidence suggesting a benefit of integrating SDT-based approaches and further suggest that nurturing intrinsic motivation can be an effective approach to supporting youth daily MVPA in under-resourced afterschool programs. TRIAL REGISTRATION: Connect Through PLAY: A Staff-based Physical Activity Intervention for Middle School Youth (Connect). https://clinicaltrials.gov/ct2/show/NCT03732144 . Registered November 6th, 2018.

The influence of social support, social affiliation and intrinsic motivation for increasing underserved youth's physical activity: A social climate-based intervention study.

The purpose of this study was to investigate the influences of intrinsic motivation, social affiliation orientations and reciprocal social support for physical activity (PA) on underserved youths' afterschool period moderate-to-vigorous (MVPA) changing trajectories across the 16-week Connect through PLAY intervention, a social-motivational climate intervention. A total of 113 youth (61.06% African American, 56.64% girls) provided full data. Youths' intrinsic motivation, social affiliation orientations and social support were measured by youth responses to a set of surveys at baseline and post-intervention. Youths' afterschool period MVPA was measured using data from 7-day ActiGraph accelerometer wear at baseline, midpoint and post-intervention. Hierarchical linear modelling analysis found that youth daily afterschool period (3pm-6pm) MVPA increased, on average, 37.94 min across the 16-week intervention. Increases in intrinsic motivation, social affiliation orientations and social support were positive predictors of youth afterschool MVPA changing trajectory. The findings clarify the contributions that a social-motivational climate intervention can have on youth afterschool period MVPA through increasing youth intrinsic motivation, social affiliation and reciprocal social support.

Cognitive Load and Energy Balance Knowledge in High-School Physical Education.

Purpose: Guided by the cognitive load theory, the purpose of this study was to determine the impacts of cognitive load and school socioeconomic status-related environmental factors on ninth-graders' energy-balanced living knowledge gain. Methods: A stratified random sample of high-school students (N = 150) participated in this study. Data were collected on students' knowledge gain, cognitive load, free and reduced-price meal rates, and student-to-teacher ratio. Results: The path analysis results revealed that the reasoning learning tasks had direct significant effects on students' knowledge gain (ßi-Diet and i-Exercise = 0.34, p < .01). The free and reduced-price meal rates and student-to-teacher ratio did not have significant effects on students' knowledge gain (p > .05). Discussion: These findings advance our understanding of the role cognitive learning tasks play in enhancing student learning in the subjects of energy-balanced knowledge and healthy lifestyle.

Learning to Teach Again: What Professional Development Approach Matters?

Purpose: Informed by the constructivist learning theory, the purpose of this study was to determine the impact of three continuing professional development (CPD) approaches on student learning in a healthful living physical education curriculum. Methods: Physical education teachers (n = 19) received one of the following CPD trainings: (a) Full Training, (b) Expedited Training, or (c) Self-Training. The effect of each CPD method was determined by tracking student learning (N = 3,418) with a two-level linear mixed model. Results: The results showed that Full Training CPD was able to generate the largest knowledge gain in both the Healthy Lifestyles Unit (ß = 0.214, p < .001) and Cardio Fitness Club Unit (ß = 0.184, p < .01) in comparison with the other two CPD approaches. Discussion: These findings advance our understanding of the role different CPD approaches play in enhancing student learning in the subjects of cardiorespiratory fitness and health lifestyles. Conclusions: The Full-Training CPD appears to benefit student learning the most followed by the Expedited-Training. The Self-Training would yield the least learning achievement.

Middle School Students' Understanding of Energy in Health and Fitness.

We used mixed methods to identify middle school students' conceptions and misconceptions of energy in the domain of health and fitness. We selected a total of 24 middle schools from six school districts in a Southeastern state of the U. S. through stratified sampling. Students were first given a standardized knowledge test to establish their knowledge level membership in the domain of health and fitness. A sample of 291 students was selected from the 24 schools for semi-structured interviews on their understanding of energy sources for physical activities and consequences of energy surplus. Analysis of the interview data identified a variety of misconceptions on energy by grade and knowledge levels. Different conceptual change theories were adopted to form four themes to explain the identified misconceptions. We discussed pedagogical implications that may help address the misconceptions in and beyond the domain of health and fitness.

The Missing Link? Middle School Students' Procedural Knowledge on Fitness.

Purpose: Guided by the declarative-procedural knowledge framework, the study attempts to identify middle school students' declarative (knowing what) and procedural (knowing how) fitness knowledge and the relationship between the two. Methods: A sample of students (n = 291, age 11-14 years) from 24 middle schools took a grade-relevant standardized knowledge test on declarative fitness knowledge and received a semistructured interview designed to clarify their declarative and procedural knowledge. Results: Most students were lacking in procedural knowledge to conduct fitness-enhancing physical activities. A few students who had mastered declarative fitness knowledge demonstrated a high level of procedural knowledge consistent with personal fitness goals. Discussion: The findings suggest that incapability to engage in fitness-enhancing physical activities could be a result of lacking procedural fitness knowledge. Future school-based interventions may prioritize procedural knowledge learning for actual physical activity participation.

Challenges in learning aerobic and anaerobic concepts: an interpretative understanding from the cognitive load theory perspective.

Purpose: It has been acknowledged in physical education (PE) that knowledge empowers learners to develop a physically active lifestyle In this study, we consider Cognitive Load Theory (CLT) as a valuable extension of the constructivist learning theory to help articulate the role of three pivotal instructional factors, content knowledge, learners, and instructional process, in learning fitness knowledge in PE. Guided by CLT, this study was intended to identify middle school students' cognitive architecture about aerobic and anaerobic concepts and pedagogical factors that might enhance or limit scientific conceptualization. Methods: We employed a mixed-methods approach. A total of 291 students randomly drawn from the 24 schools provided data on conceptual knowledge levels via standardized test and conceptual structures of understanding via semi-structured interviews about the aerobic/anaerobic concepts. Results: Results from integrated quantitative and qualitative analyses reveal the role of task element interactivity as a function of conceptualization in the students' cognitive architecture. The students lack the understanding that aerobic and anaerobic concepts are associated with the concepts of complex metabolic systems that dictate the energy pathways experienced in physical activity. Their understandings are characterized by a tendency of overly simplifying the complex conceptions for convenient schema activation. The simplified conception is likely to prohibit their understanding from becoming consistent with the science. The findings suggest that teachers should anticipate lack of understanding of knowledge connectivity when teaching fitness concepts and establish the task element interactivity with enhanced clarity to facilitate knowledge learning. Conclusion: Consistent with the constructivist learning theory, the findings inform us that teaching complex fitness concepts require the teacher to connect prior knowledge as a prerequisite to activate relevant cognitive architecture and actively build connections between the prior knowledge and the to-be learned concepts. PE curriculum designers and teachers should anticipate the complex challenges in teaching the aerobic and anaerobic concepts because of their connections to other concepts associated with human energy systems such as intensity and oxygen supply.

Impact of elevated Ca(2+)/Mg(2+) concentrations of reverse osmosis membrane desalinated seawater on the stability of water pipe materials.

Hardness and alkalinity are known factors influencing the chemical stability of desalinated water. This study was carried out to investigate the effect of Ca(2+) and Mg(2+) on corrosion and/or scale formation on the surface of different water distribution pipe materials under tropical conditions. The corrosion rates of ductile iron, cast iron and cement-lined ductile iron coupons were examined in reverse osmosis (RO) membrane desalinated seawater which was remineralised using different concentrations of Ca(2+) and Mg(2+). The changes in water characteristics and the coupon corrosion rates were studied before and after the post-treatment. The corrosion mechanisms and corrosion products were examined using scanning electron microscope and X-ray diffraction, respectively. We found that the combination of Ca(2+) and Mg(2+) (60/40 mg/L as CaCO3) resulted in lower corrosion rates than all other treatments for the three types of pipe materials, suggesting that Ca(2+)/Mg(2+) combination improves the chemical stability of desalinated seawater rather than Ca(2+) only.

An amphioxus RAG1-like DNA fragment encodes a functional central domain of vertebrate core RAG1.

The highly diversified repertoire of antigen receptors in the vertebrate immune system is generated via proteins encoded by the recombination activating genes (RAGs) RAG1 and RAG2 by a process known as variable, diversity, and joining [V(D)J] gene recombination. Based on the study of vertebrate RAG proteins, many hypotheses have been proposed regarding the origin and evolution of RAG. This issue remains unresolved, leaving a significant gap in our understanding of the evolution of adaptive immunity. Here, we show that the amphioxus genome contains an ancient RAG1-like DNA fragment (bfRAG1L) that encodes a virus-related protein that is much shorter than vertebrate RAG1 and harbors a region homologous to the central domain of core RAG1 (cRAG1). bfRAG1L also contains an unexpected retroviral type II nuclease active site motif, DXN(D/E)XK, and is capable of degrading both DNA and RNA. Moreover, bfRAG1L shares important functional properties with the central domain of cRAG1, including interaction with RAG2 and localization to the nucleus. Remarkably, the reconstitution of bfRAG1L into a cRAG1-like protein yielded an enzyme capable of recognizing recombination signal sequences and performing V(D)J recombination in the presence of mouse RAG2. Moreover, this reconstituted cRAG1-like protein could mediate the assembly of antigen receptor genes in RAG1-deficient mice. Together, our results demonstrate that amphioxus bfRAG1L encodes a protein that is functionally equivalent to the central domain of cRAG1 and is well prepared for further evolution to mediate V(D)J recombination. Thus, our findings provide unique insights into the evolutionary origin of RAG1.

Activator protein 1 suppresses antitumor T-cell function via the induction of programmed death 1.

T cells play a critical role in tumor immunosurveillance by eliminating newly transformed somatic cells. However, tumor cell variants can escape from immunological control after immunoediting, leading to tumor progression. Whether and how T cells respond to tumor growth remain unclear. Here, we found that tumor-infiltrating T cells exhibited persistently up-regulated expression of the activator protein 1 (AP-1) subunit c-Fos during tumor progression. The ectopic expression of c-Fos in T cells exacerbated tumor growth, whereas the T-cell-specific deletion of c-Fos reduced tumor malignancy. This unexpected immunosuppressive effect of c-Fos was mediated through the induced expression of immune inhibitory receptor programmed death 1 (PD-1) via the direct binding of c-Fos to the AP-1-binding site in the Pdcd1 (gene encoding PD-1) promoter. A knock-in mutation of this binding site abrogated PD-1 induction, augmented antitumor T-cell function and repressed tumor growth. Taken together, these findings indicate that T-cell c-Fos subsequently induces PD-1 expression in response to tumor progression and that disrupting such induction is essential for repression of tumor growth.