A Toxoplasma gondii thioredoxin with cell adhesion and antioxidant function.

Background: Toxoplasma gondii (T. gondii) is a widespread, zoonotic protozoan intracellular parasite with a complex life cycle, which can cause toxoplasmosis, a potentially serious disease. During the invasion process, T. gondii proteins first bind to the relevant host cell receptors, such as glycosaminoglycan molecule (GAG-binding motif), which is one of the main receptors for parasites or virus to infect host cells. However, research on TGME49_216510 (T. gondii Trx21), a protein from Toxoplasma gondii, is limited. Methods: Bioinformatics analysis of the Trx21 protein was performed firstly. And specific primers were then designed using the conserved domain and GAG-binding motif to amplify, express, and purify a fragment of the Trx21 protein. The purified Trx21-GST protein was used for antioxidant and cell adhesion experiments. Simultaneously, mice were immunized with Trx21-His to generate specific polyclonal antibodies for subcellular localization analysis. Results: The Trx21 protein, consisting of 774 amino acids, included a transmembrane region, three GAG-binding motifs, and a Thioredoxin-like domain. The recombinant Trx21-His protein had a molecular mass of about 31 kDa, while the Trx21-GST protein had a molecular mass of about 55 kDa, which was analyzed by SDS-PAGE and Western blot. Subcellular localization analysis by IFA revealed that Trx21 is predominantly distributed in the cytoplasm of T. gondii. Furthermore, Trx21 exhibited a protective effect on supercoiled DNA against metal-catalyzed oxidation (MCO) and demonstrated adhesion abilities to Vero cells. Conclusions: These results indicate that Trx21 plays an important role in host cell interaction and oxidative damage.

Which horticultural activities are more effective for children's recovery from stress and mental fatigue? A quasi-experimental study.

Introduction: Studies have established the benefits of horticultural therapy and activities for human health and well-being. Nonetheless, limited research has been conducted on the potential restorative advantages and distinctions between different types of horticultural activities in terms of stress reduction. Methods: This study employed a quantitative research method to investigate the stress recovery benefits of five horticultural activities (flower arrangement, sowing and transplanting seeds, kokedama crafting, pressed flower card making, and decorative bottle painting with dried flowers) and one reference activity (short composition writing) for children. The experiment was conducted in a children's activity center's multi-purpose classroom with 48 elementary students aged 9-12 years. The subjects first took a stress test to induce stress and then engaged in horticultural activities for 20 min. Physiological stress was assessed using electrocardiograms and electroencephalograms as feedback indicators. Psychological and emotional changes were determined using the Positive and Negative Affect Schedule for Children and Self-Assessment Manikin scales. Results: The results demonstrated that horticultural activities greatly reduced physiological fatigue, and their recovery benefits were significantly greater than those of the reference activity. The recovery effects from different horticultural activities were similar across physiological indicators, although flower arrangement and sowing and transplanting seeds exhibited relatively robust recovery benefits. The heart rate and α-EEG-based generalized estimating equation revealed that horticultural activities offered significantly better relative recovery at each time phase of operation than the reference activity, with girls showing a 3.68% higher relative recovery value than boys. Flower arrangement and kokedama crafting offered better physiological recovery for students with prior horticultural experience, and these two activities received the highest scores in terms of positive effects and the "pleasure" dimension. Students believed that participating in horticultural activities resulted in a noteworthy increase in personal confidence and a greater sense of achievement. Conclusion: The study suggests that horticultural activities that involve real and vibrant plants or natural materials and are more attractive have more stress-relieving benefits. We conclude that horticultural activities are beneficial leisure activities that aid in stress relief for children and that it is important to consider the attributes of activities when developing horticultural programs for elementary students.

Comparative analysis of codon usage patterns of Plasmodium helical interspersed subtelomeric (PHIST) proteins.

Background: Plasmodium falciparum is a protozoan parasite that causes the most severe form of malaria in humans worldwide, which is predominantly found in sub-Saharan Africa, where it is responsible for the majority of malaria-related deaths. Plasmodium helical interspersed subtelomeric (PHIST) proteins are a family of proteins, with a conserved PHIST domain, which are typically located at the subtelomeric regions of the Plasmodium falciparum chromosomes and play crucial roles in the interaction between the parasite and its human host, such as cytoadherence, immune evasion, and host cell remodeling. However, the specific utilization of synonymous codons by PHIST proteins in Plasmodium falciparum is still unknown. Methods: Codon usage bias (CUB) refers to the unequal usage of synonymous codons during translation, resulting in over- or underrepresentation of certain nucleotide patterns. This imbalance in CUB can impact various cellular processes, including protein expression levels and genetic variation. To investigate this, the CUB of 88 PHIST protein coding sequences (CDSs) from 5 subgroups were analyzed in this study. Results: The results showed that both codon base composition and relative synonymous codon usage (RSCU) analysis identified a higher occurrence of AT-ended codons (AGA and UUA) in PHIST proteins of Plasmodium falciparum. The average effective number of codons (ENC) for these PHIST proteins was 36.69, indicating a weak codon preference among them, as it was greater than 35. Additionally, the correlation analysis among codon base composition (GC1, GC2, GC3, GCs), codon adaptation index (CAI), codon bias index (CBI), frequency of optimal codons (FOP), ENC, general average hydropathicity (GRAVY), aromaticity (AROMO), length of synonymous codons (L_sym), and length of amino acids (L_aa) revealed the influence of base composition and codon usage indices on codon usage bias, with GC1 having a significant impact in this study. Furthermore, the neutrality plot analysis, PR2-bias plot analysis, and ENC-GC3 plot analysis provided additional evidence that natural selection plays a crucial role in determining codon bias in PHIST proteins. Conclusion: In conclusion, this study has enhanced our understanding of the characteristics of codon usage and genetic evolution in PHIST proteins, thereby providing data foundation for further research on antimalarial drugs or vaccines.

Association of red cell distribution width and its changes with the 30-day mortality in patients with acute respiratory failure: An analysis of MIMIC-IV database.

BACKGROUND: Acute respiratory failure (ARF) is a common disease in the intensive care units (ICUs) with high risk of mortality. The red cell distribution width (RDW) is one of baseline ICU indicators which can be easily available, and has been used in the long-term prognostic analyses of diseases. However, no studies have explored the role of baseline RDW and its change during hospitalization in in-hospital mortality in ARF. Herein, this study aims to explore the association between RDW and its changes and the 30-day mortality in ARF patients. METHODS: Demographic and clinical data of 7,497 patients with ARF were extracted from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database in 2012-2019 in this retrospective cohort study. Univariable and multivariable Cox regression analyses were used to explore the association between RDW and its changes and 30-day mortality with hazard ratios (HRs) and 95% confidence intervals (CIs). Subgroup analyses of different baseline RDW levels were also performed. We then assessed the predictive performance of RDW changes combined with the Sequential Organ Failure Assessment (SOFA) score on 30-day mortality using receiver operator characteristic curves (ROCs) with areas under curve (AUCs). RESULTS: Totally, 2,254 (30.07%) patients died in 30 days. After adjusting for covariates, we found that high baseline RDW [HR = 1.25, 95%CI: (1.15-1.37)] and RDW changes ≥0.3% [HR = 1.12, 95%CI: (1.01-1.24)] were both related to an increased risk of 30-day mortality. In patients whose baseline RDW level ≥14.9%, RDW changes ≥0.3% was also associated with an increased risk of 30-day mortality [HR = 1.19, 95%CI: (1.05-1.35)]. Moreover, the predictive value of RDW changes combined with SOFA on 30-day mortality was a little better than that of single SOFA score, with AUCs of 0.624 vs. 0.620. CONCLUSION: High baseline RDW level and its changes during hospitalization was relate to the increased risk of 30-day mortality in ARF, and the predictive value of RDW changes for ARF short-term mortality is still needed exploration.

LncRNA NKILA Promotes Epithelial-Mesenchymal Transition of Liver Cancer Cells by Targeting miR-485-5p.

OBJECTIVE: Liver cancer (LC), one of the familiar malignancies, has a very high morbidity all over the world. The onset of the disease is hidden, and the patients usually do not express any special symptoms. Most of them will have been developed to the middle and later stage when they are diagnosed. This is one of the main reasons why the prognosis of LC is extremely pessimistic all the year round. Recently, researchers have focused mainly on molecular studies, among which LncRNA is a hot spot. This research aims to explore the biological behaviors of LncRNA NKILA and miR-485-5p in LC cells and verify the relationship between them, thereby providing a new theoretical basis for future prevention and treatment. METHODS: Ninety-four early LC patients admitted to our hospital from January 2015 to January 2017 were regarded as the research objects. In addition, human LC cells SMMC-7721, HepG2, and normal liver cells HL-7702 were purchased. The LncRNA NKILA and miR-485-5p level in cancer and adjacent tissues, LC, and normal liver cells of patients was tested by PCR. Patients were followed up for 3 years. Then, LncRNA NKILA and miR-485-5p's effects on prognosis and cell biological behavior were analyzed. At last, the relationship between LncRNA NKILA and miR-485-5p was assessed by a dual-luciferase reporter assay. RESULTS: The LncRNA NKILA expression was high in LC tissues and cells (P < 0.050), while miR-485-5p was low compared with the normal adjacent tissues (P < 0.050). Prognostic follow-up manifested that high LncRNA NKILA or low miR-485-5p could predict the poor prognosis and high mortality risk of the patients (P < 0.050). LC cells with downregulated LncRNA NKILA documented inhibited proliferation, invasion, and EMT, while the apoptosis level of the cells increased (P < 0.050). The proliferation, invasion, and EMT were inhibited by miR-485-5p increase, while the apoptosis of the cells decreased after upregulating miR-485-5p (P < 0.050). Online websites predicted that LncRNA NKILA had a binding site with miR-485-5p, and dual-luciferase reporter assay confirmed that LncRNA NKILA could directly target with miR-485-5p (P < 0.050). The miR-485-5p in LC cells increased after LncRNA NKILA was silenced (P < 0.050). The rescue experiment documented that LncRNA NKILA inhibition on LC cells was reversed by inhibiting miR-485-5p (P < 0.050). CONCLUSION: The LncRNA NKILA with high expression advances LC cell proliferation, invasion, and EMT by targeting miR-485-5p.