Bioorthogonal Delivery of Carbon Disulfide in Living Cells.

Carbon disulfide (CS2) is an environmental pollutant, which is deadly hazardous to the workers under chronic or acute exposure. However, the toxicity mechanisms of CS2 are still unclear due to the scarcity of biocompatible donors, which can release CS2 in cells. Here we developed the first bioorthogonal CS2 delivery system based on the "click-and-release" reactions between mesoionic 1,3-thiazolium-5-thiolates (TATs) and strained cyclooctyne exo-BCN-OH. We successfully realized intracellular CS2 releasing and investigated the causes of CS2-induced hepatotoxicity, including oxidative stress, proteotoxic stress and copper-dependent cell death. It is found that CS2 can be copper vehicles bypassing copper transporters after reacting with nucleophiles in cytoplasm, and extra copper supplementation will exacerbate the loss of homeostasis of cells and ultimately cell death. These findings inspired us to explore the anticancer activity of CS2 in combination with copper by introducing a copper chelating group in our CS2 delivery system.

The hyphal-specific toxin candidalysin promotes fungal gut commensalism.

The fungus Candida albicans frequently colonizes the human gastrointestinal tract, from which it can disseminate to cause systemic disease. This polymorphic species can transition between growing as single-celled yeast and as multicellular hyphae to adapt to its environment. The current dogma of C. albicans commensalism is that the yeast form is optimal for gut colonization, whereas hyphal cells are detrimental to colonization but critical for virulence1-3. Here, we reveal that this paradigm does not apply to multi-kingdom communities in which a complex interplay between fungal morphology and bacteria dictates C. albicans fitness. Thus, whereas yeast-locked cells outcompete wild-type cells when gut bacteria are absent or depleted by antibiotics, hyphae-competent wild-type cells outcompete yeast-locked cells in hosts with replete bacterial populations. This increased fitness of wild-type cells involves the production of hyphal-specific factors including the toxin candidalysin4,5, which promotes the establishment of colonization. At later time points, adaptive immunity is engaged, and intestinal immunoglobulin A preferentially selects against hyphal cells1,6. Hyphal morphotypes are thus under both positive and negative selective pressures in the gut. Our study further shows that candidalysin has a direct inhibitory effect on bacterial species, including limiting their metabolic output. We therefore propose that C. albicans has evolved hyphal-specific factors, including candidalysin, to better compete with bacterial species in the intestinal niche.

Chemoenzymatic Installation of Site-Specific Chemical Groups on DNA Enhances the Catalytic Activity.

Functional DNAs are valuable molecular tools in chemical biology and analytical chemistry but suffer from low activities due to their limited chemical functionalities. Here, we present a chemoenzymatic method for site-specific installation of diverse functional groups on DNA, and showcase the application of this method to enhance the catalytic activity of a DNA catalyst. Through chemoenzymatic introduction of distinct chemical groups, such as hydroxyl, carboxyl, and benzyl, at specific positions, we achieve significant enhancements in the catalytic activity of the RNA-cleaving deoxyribozyme 10-23. A single carboxyl modification results in a 100-fold increase, while dual modifications (carboxyl and benzyl) yield an approximately 700-fold increase in activity when an RNA cleavage reaction is catalyzed on a DNA-RNA chimeric substrate. The resulting dually modified DNA catalyst, CaBn, exhibits a kobs of 3.76 min-1 in the presence of 1 mM Mg2+ and can be employed for fluorescent imaging of intracellular magnesium ions. Molecular dynamics simulations reveal the superior capability of CaBn to recruit magnesium ions to metal-ion-binding site 2 and adopt a catalytically competent conformation. Our work provides a broadly accessible strategy for DNA functionalization with diverse chemical modifications, and CaBn offers a highly active DNA catalyst with immense potential in chemistry and biotechnology.

Breast Cancer Knowledge and Mammography Use Among Asian American Women Aged 40 and Older: Using the Transtheoretical Model Approach.

Mammography screening rates remain low among Asian American women (AAW). The aims of our study were to: (a) assess breast cancer knowledge and mammography screening behaviors, and (b) identify the factors related to the transtheoretical model (TTM) stages of change in relation to mammography utilization among AAW aged 40 and older. Using a cross-sectional design, a convenience sample of 714 AAW completed a structured questionnaire in 2021. Participants demonstrated a moderate level of knowledge regarding breast cancer and mammography. Only 34.2% of the participants reported obtaining regular mammograms. The ordinal logistic regression indicated that age, birthplace, health perception, breast biopsy history, breast cancer knowledge, self-efficacy, and perceived barriers were correlated with TTM stages of change. Our results highlight the need for implementing effective interventions aimed at increasing knowledge and screening rates for breast cancer among AAW. Additional TTM studies with AAW are needed to determine the relationships among TTM constructs and develop theory-based programs to improve adherence to screening guidelines. Future research using a mixed-method design may provide opportunities to explore complex phenomena associated with breast cancer screening behaviors. Finally, further assessments of the Breast Cancer Knowledge Scale's psychometric properties are necessary to improve this instrument.

Human papillomavirus vaccination: a quantitative cross-sectional study of perceived barriers, influential advisors, and acculturation among Chinese college students aged 18-26 in the USA.

HPV vaccines are highly effective in preventing HPV-associated cancers; however, HPV vaccination uptake is low among Chinese students studying at U.S. colleges. The purposes of this study were to evaluate (a) perceived barriers and influential others trusted for advice regarding HPV vaccination and (b) factors (i.e. HPV vaccination, acculturation) related to barriers and influential advisors among 18- to 26-year-old Chinese students attending U.S. colleges. We used a cross-sectional design to obtain self-reported data in 2019 from a chain-referral sample of 213 Chinese students. Among 125 respondents who were unvaccinated or partially vaccinated, the reported barriers to receiving the HPV vaccine included: (a) lack of recommendations from a healthcare provider, (b) lack of risk perception for HPV infection, and (c) limited knowledge about HPV vaccination locations. The influential advisors for receiving HPV vaccination were doctors, parents, self, nurses, and same-sex friends. Multivariate analysis revealed that unvaccinated respondents were more likely to report the following barriers to HPV vaccination: (a) lack of recommendations from a healthcare provider, (b) lack of risk perception for HPV infection, (c) limited knowledge about vaccination locations, and (d) uncertainty about effectiveness. High Asian identified respondents were more likely to perceive barriers related to limited knowledge about vaccination locations and uncertainty about effectiveness, while they were less likely to state nurses as influential advisors. Individuals who received one or more HPV vaccine doses were more inclined to view same-sex friends and nurses as influential advisors for HPV vaccination. The influence of culture on preferences for information sources, such as specific providers and provider gender, needs to be addressed. Programs designed to decrease barriers and improve HPV vaccination among Chinese students should also focus on acculturation status.

Spatiotemporal-social association predicts immunological similarity in rewilded mice.

Environmental influences on immune phenotypes are well-documented, but our understanding of which elements of the environment affect immune systems, and how, remains vague. Behaviors, including socializing with others, are central to an individual's interaction with its environment. We therefore tracked behavior of rewilded laboratory mice of three inbred strains in outdoor enclosures and examined contributions of behavior, including associations measured from spatiotemporal co-occurrences, to immune phenotypes. We found extensive variation in individual and social behavior among and within mouse strains upon rewilding. In addition, we found that the more associated two individuals were, the more similar their immune phenotypes were. Spatiotemporal association was particularly predictive of similar memory T and B cell profiles and was more influential than sibling relationships or shared infection status. These results highlight the importance of shared spatiotemporal activity patterns and/or social networks for immune phenotype and suggest potential immunological correlates of social life.

Composite Dietary Antioxidant Index Is Negatively Associated with Hyperuricemia in US Adults: An Analysis of NHANES 2007-2018.

Hyperuricemia and its complications are severe risks to human health. Dietary intervention is considered an essential part of the management of hyperuricemia. Studies have reported that the intake of antioxidants has a positive effect on hyperuricemia. Here, we collected data from 8761 participants of the National Health and Nutrition Examination Survey for this analysis. Daily intakes of vitamins A, C, and E; manganese; selenium; and zinc were calculated as the composite dietary antioxidant index (CDAI). The participants were divided into four groups (Q1, Q2, Q3, and Q4) according to the CDAI. Univariate analysis was used to assess the association of covariates with hyperuricemia. The association between the CDAI and hyperuricemia was evaluated using multinomial logistic regression, and its stability was determined by stratified analysis. Our results revealed that the CDAI has a significant negative association with hyperuricemia (Q2: 0.81 (0.69, 0.95); Q3: 0.75 (0.62, 0.90); Q4: 0.65 (0.51, 0.82); P < 0.01). The results of stratified analysis emphasize that this association between CDAI and hyperuricemia is stable. In conclusion, this study suggested a negative association between the CDAI and hyperuricemia.

Antimicrobial overproduction sustains intestinal inflammation by inhibiting Enterococcus colonization.

Loss of antimicrobial proteins such as REG3 family members compromises the integrity of the intestinal barrier. Here, we demonstrate that overproduction of REG3 proteins can also be detrimental by reducing a protective species in the microbiota. Patients with inflammatory bowel disease (IBD) experiencing flares displayed heightened levels of secreted REG3 proteins that mediated depletion of Enterococcus faecium (Efm) from the gut microbiota. Efm inoculation of mice ameliorated intestinal inflammation through activation of the innate immune receptor NOD2, which was associated with the bacterial DL-endopeptidase SagA that generates NOD2-stimulating muropeptides. NOD2 activation in myeloid cells induced interleukin-1ß (IL-1ß) secretion to increase the proportion of IL-22-producing CD4+ T helper cells and innate lymphoid cells that promote tissue repair. Finally, Efm was unable to protect mice carrying a NOD2 gene variant commonly found in IBD patients. Our findings demonstrate that inflammation self-perpetuates by causing aberrant antimicrobial activity that disrupts symbiotic relationships with gut microbes.

Differentiation, regulation and function of regulatory T cells in non-lymphoid tissues and tumors.

Regulatory T cells (Tregs) play a substantial role in inhibiting excessive immune response. A large number of studies have focused on the tissue homeostasis maintenance and remodeling characteristics of Tregs in non-lymphoid tissues, such as the skin, colon, lung, brain, muscle, and adipose tissues. Herein, we overview the kinetics of Treg migration to non-lymphoid tissues and adaptation to the specific tissue microenvironment through the development of tissue-specific chemokine receptors, transcription factors, and phenotypes. Additionally, tumor-infiltrating Tregs (Ti-Tregs) play an important role in tumor generation and immunotherapy resistance. The phenotypes of Ti-Tregs are related to the histological location of the tumor and there is a large overlap between the transcripts of Ti-Tregs and those of tissue-specific Tregs. We recapitulate the molecular underpinnings of tissue-specific Tregs, which might shed new light on Treg-based therapeutic targets and biomarkers for inflammatory diseases and cancer.

Site-Selective Arylation of Carboxamides from Unprotected Peptides.

The amidated peptides are an important class of biologically active compounds due to their unique biological properties and wide applications as potential peptide drugs and biomarkers. Despite the abundance of free amide motifs (Asn, Gln, and C-terminal amide) in native peptides, late-stage modification of the amide unit in naturally occurring peptides remains very rare because of the intrinsically weak nucleophilicity of amides and the interference of multiple competing nucleophilic residues, which generally lead to undesired side reactions. Herein, chemoselective arylation of amides in unprotected polypeptides has been developed under an air atmosphere to afford the N-aryl amide peptides bearing various functional motifs. Its success relies on the combination of gold catalysis and silver salt to differentiate the relative inert amide among a collection of reactive nucleophilic amino acid residues (e.g., -NH2, -OH, and -COOH), favoring the C-N bond coupling toward amides over other more nucleophilic groups. Experimental and DFT studies reveal a crucial role of the silver cation, which serves as a transient coordination mask of the more reactive reaction sites, overcoming the inherently low reactivity of amides. The excellent biocompatibility of this strategy has been applied to functionalize a wide range of peptide drugs and complex peptides. The application could be further extended to peptide labeling and peptide stapling.

Rewilding of laboratory mice enhances granulopoiesis and immunity through intestinal fungal colonization.

The paucity of blood granulocyte populations such as neutrophils in laboratory mice is a notable difference between this model organism and humans, but the cause of this species-specific difference is unclear. We previously demonstrated that laboratory mice released into a seminatural environment, referred to as rewilding, display an increase in blood granulocytes that is associated with expansion of fungi in the gut microbiota. Here, we find that tonic signals from fungal colonization induce sustained granulopoiesis through a mechanism distinct from emergency granulopoiesis, leading to a prolonged expansion of circulating neutrophils that promotes immunity. Fungal colonization after either rewilding or oral inoculation of laboratory mice with Candida albicans induced persistent expansion of myeloid progenitors in the bone marrow. This increase in granulopoiesis conferred greater long-term protection from bloodstream infection by gram-positive bacteria than by the trained immune response evoked by transient exposure to the fungal cell wall component ß-glucan. Consequently, introducing fungi into laboratory mice may restore aspects of leukocyte development and provide a better model for humans and free-living mammals that are constantly exposed to environmental fungi.

Asymmetric iminium ion-catalyzed conjugate addition of 2-hydroxycinnamaldehydes and 2-oxocarboxylic esters: synthesis of chiral polysubstituted bridged bicyclic ketals.

A highly regio-, chemo-, and stereoselective cascade process initiated by enantioselective iminium-catalyzed conjugate addition of 2-hydroxycinnamaldehydes and 2-oxocarboxylic esters is presented. Normal cinnamaldehydes are not reactive under the same reaction conditions. Bridged bicyclic ketals rather than acetals bearing stereocenters on both the bridge carbon and bridgehead ketal carbon are synthesized.

L-Fucose increases the fucosylation of colorectal cancer cells via promoting the accumulation of serine.

Fucosylation, a kind of posttranslational modification, has been identified as a key regulator of health, with alterations in this process serving as an indicator of diseases such as colorectal cancer. L-Fucose, an essential substrate of fucosylation, was reported to possess anticancer potential and increase fucosylation. However, the association between its tumour-inhibitory effect and its ability to regulate fucosylation was not fully understood. Herein, we demonstrate that the simultaneous inhibitory effect of L-fucose on cancer cell growth and enhanced fucosylation occurred only in certain colorectal cancer cells (HCT-116 cells) but not in normal cells (HCoEpic cells), which may be related to the induction of pro-apoptotic fucosylated proteins by L-fucose in HCT-116 cells. RNA-seq analysis showed that upregulation of the transcription levels of serine biosynthesis genes (e.g. PSAT1) and decreased levels of genes involved in serine consumption with supplemental L-fucose were also unique to HCT-116 cells. Increased serine concentrations only in HCT-116 cells and increased α1,3/6-fucosylation in CRC cells induced by exogenous serine also verified that L-fucose enhanced fucosylation via promoting intracellular serine accumulation. Additionally, the knockdown of PSAT1 and serine-deficiency impaired fucosylation. Notably, PSAT1 knockdown weakened the inhibitory effect of L-fucose on cell proliferation and migration. Interestingly, simultaneous increased levels of α1,3/6-fucosylation and PSAT1 transcription were also identified in colorectal tumor tissues of CRC patients. Together, these results uncover a novel role of serine synthesis and PSAT1 in the regulation of fucosylation and provide insights into the potential application of L-fucose in CRC therapy.

Genetic and Environmental interactions contribute to immune variation in rewilded mice.

The relative and synergistic contributions of genetics and environment to inter-individual immune response variation remain unclear, despite its implications for understanding both evolutionary biology and medicine. Here, we quantify interactive effects of genotype and environment on immune traits by investigating three inbred mouse strains rewilded in an outdoor enclosure and infected with the parasite, Trichuris muris. Whereas cytokine response heterogeneity was primarily driven by genotype, cellular composition heterogeneity was shaped by interactions between genotype and environment. Notably, genetic differences under laboratory conditions can be decreased following rewilding, and variation in T cell markers are more driven by genetics, whereas B cell markers are driven more by environment. Importantly, variation in worm burden is associated with measures of immune variation, as well as genetics and environment. These results indicate that nonheritable influences interact with genetic factors to shape immune variation, with synergistic impacts on the deployment and evolution of defense mechanisms.

Antimicrobial overproduction sustains intestinal inflammation by inhibiting Enterococcus colonization.

Loss of antimicrobial proteins such as REG3 family members compromises the integrity of the intestinal barrier. Here, we demonstrate that overproduction of REG3 proteins can also be detrimental by reducing a protective species in the microbiota. Patients with inflammatory bowel disease (IBD) experiencing flares displayed heightened levels of secreted REG3 proteins that mediated depletion of Enterococcus faecium ( Efm ) from the gut microbiota. Efm inoculation of mice ameliorated intestinal inflammation through activation of the innate immune receptor NOD2, which was associated with the bacterial DL-endopeptidase SagA. Microbiota sensing by NOD2 in myeloid cells mediated IL-1ß secretion and increased the proportion of IL-22-producing CD4 + T helper cells and innate lymphoid cells. Finally, Efm was unable to protect mice carrying a NOD2 gene variant commonly found in IBD patients. Our findings demonstrate that inflammation self-perpetuates by causing aberrant antimicrobial activity that disrupts symbiotic relationships with gut microbes.

Marine-Derived Natural Product HDYL-GQQ-495 Targets P62 to Inhibit Autophagy.

Autophagy is widely implicated in pathophysiological processes such as tumors and metabolic and neurodegenerative disorders, making it an attractive target for drug discovery. Several chemical screening approaches have been developed to uncover autophagy-modulating compounds. However, the modulation capacity of marine compounds with significant pharmacological activities is largely unknown. We constructed an EGFPKI-LC3B cell line using the CRISPR/Cas9 knock-in strategy in which green fluorescence indicated endogenous autophagy regulation. Using this cell line, we screened a compound library of approximately 500 marine natural products and analogues to investigate molecules that altered the EGFP fluorescence. We identified eight potential candidates that enhanced EGFP fluorescence, and HDYL-GQQ-495 was the leading one. Further validation with immunoblotting demonstrated that cleaved LC3 was increased in dose- and time-dependent manners, and the autophagy adaptor P62 showed oligomerization after HDYL-GQQ-495 treatment. We also demonstrated that HDYL-GQQ-495 treatment caused autophagy substrate aggregation, which indicated that HDYL-GQQ-495 serves as an autophagy inhibitor. Furthermore, HDYL-GQQ-495 induced Gasdermin E (GSDME) cleavage and promoted pyroptosis. Moreover, HDYL-GQQ-495 directly combined with P62 to induce P62 polymerization. In P62 knockout cells, the cleavage of LC3 or GSDME was blocked after HDYL-GQQ-495 treatment. The EGFPKI-LC3B cell line was an effective tool for autophagy modulator screening. Using this tool, we found a novel marine-derived compound, HDYL-GQQ-495, targeting P62 to inhibit autophagy and promote pyroptosis.

Bayesian Inference for an Unknown Number of Attributes in Restricted Latent Class Models.

The specification of the [Formula: see text] matrix in cognitive diagnosis models is important for correct classification of attribute profiles. Researchers have proposed many methods for estimation and validation of the data-driven [Formula: see text] matrices. However, inference of the number of attributes in the general restricted latent class model remains an open question. We propose a Bayesian framework for general restricted latent class models and use the spike-and-slab prior to avoid the computation issues caused by the varying dimensions of model parameters associated with the number of attributes, K. We develop an efficient Metropolis-within-Gibbs algorithm to estimate K and the corresponding [Formula: see text] matrix simultaneously. The proposed algorithm uses the stick-breaking construction to mimic an Indian buffet process and employs a novel Metropolis-Hastings transition step to encourage exploring the sample space associated with different values of K. We evaluate the performance of the proposed method through a simulation study under different model specifications and apply the method to a real data set related to a fluid intelligence matrix reasoning test.

Optimization of the Beijing Economy-Energy-Emissions System 2021-2035: A Scenario Simulation Analysis Based on a System Dynamics Model.

This paper constructs an Economy-Energy-Emissions (3E) System Dynamics Model using the megacity of Beijing, China, as an example, to estimate the effects of different policy scenarios (including three single-policy scenarios and four combined-policy scenarios) on the core variables of Beijing's 3E system from 2021 to 2035. The results suggest two main points. (1) Following the current development trend, the proportion of the GDP represented by the added value of advanced high-precision industries (Gao Jing Jian in Chinese) will only be 43% in 2035, implying a limited role in promoting economic growth. Despite effective control of total energy consumption, fossil energy's share of total consumption will reach 57% by 2035, hindering the process of making the energy consumption structure cleaner and leading to failure to achieve the targeted inflection point in CO2 emissions by 2025. PM2.5 control shows some successful results and will decrease to 19 µg/m3 in 2035. However, a gap compared to other world-class cities remains. (2) The implementation of a single policy for either industrial structure optimization, energy structure transformation, or emissions control cannot simultaneously meet the goal of high-quality coordinated development of Beijing's 3E system, whereas the comprehensive implementation of policies in all three dimensions is demonstrably effective.