Publisher Correction: An efficient gene knock-in strategy using 5'-modified double-stranded DNA donors with short homology arms.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

An efficient gene knock-in strategy using 5'-modified double-stranded DNA donors with short homology arms.

Here, we report a rapid CRISPR-Cas9-mediated gene knock-in strategy that uses Cas9 ribonucleoprotein and 5'-modified double-stranded DNA donors with 50-base-pair homology arms and achieved unprecedented 65/40% knock-in rates for 0.7/2.5 kilobase inserts, respectively, in human embryonic kidney 293T cells. The identified 5'-end modification led to up to a fivefold increase in gene knock-in rates at various genomic loci in human cancer and stem cells.

Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli.

Most native producers of ribosomally synthesized and post-translationally modified peptides (RiPPs) utilize N-terminal leader peptides to avoid potential cytotoxicity of mature products to the hosts. Unfortunately, the native machinery of leader peptide removal is often difficult to reconstitute in heterologous hosts. Here we devised a general method to produce bioactive lanthipeptides, a major class of RiPP molecules, in Escherichia coli colonies using synthetic biology principles, where leader peptide removal is programmed temporally by protease compartmentalization and inducible cell autolysis. We demonstrated the method for producing two lantibiotics, haloduracin and lacticin 481, and performed analog screening for haloduracin. This method enables facile, high throughput discovery, characterization, and engineering of RiPPs.

Association of Interdental Cleaning and Untreated Root Caries in Adults in the United States of America.

PURPOSE: This study aimed to evaluate the association between interdental cleaning and untreated root caries amongst middle-aged and older adults in the US. MATERIALS AND METHODS: Data were obtained from the National Health and Nutrition Examination Survey (NHANES) (2015-2016 and 2017-2018). Adults aged ≥40 years who underwent full mouth examination and root caries assessment were included. Participants were classified based on their interdental cleaning frequency as none, 1-3 d/wk, and 4-7 d/wk. Associations between interdental cleaning and untreated root caries were assessed using a weighted multivariable logistic regression model adjusted for sociodemographics, general behaviour, general health condition, oral conditions, oral health behavior, and dietary factors. Subgroup analyses stratified by age and sex were performed after adjusting for covariates in the logistic regression models. RESULTS: The prevalence of untreated root caries was 15.3% amongst 6217 participants. Interdental cleaning for 4-7 d/wk was a significant risk factor (odds ratio, 0.67; 95% confidence interval, 0.52-0.85). It was associated with a 40% reduction in the risk of untreated root caries in participants aged 40 to 64 years and a 37% reduction in women. Untreated root caries was also significantly associated with age, family income, smoking status, root restoration, number of teeth, untreated coronal caries, and recent dental visit. CONCLUSIONS: Interdental cleaning for 4-7 d/wk was associated with fewer untreated root caries amongst middle-aged adults and women in the US. The risk of root caries increases with age. Low family income was a risk indicator for root caries amongst middle-aged adults. Additionally, smoking, root restoration, number of teeth, untreated coronal caries, and recent dental visits were common risk factors for root caries in middle-aged and older people in the US.

Rare Trichilemmal Cyst Localized in the Pulp of the Middle Finger.

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[Health Benefit Evaluation for PM2.5 as Well as O3-8h Pollution Control in Chengdu, China from 2016 to 2020].

Both particulate matter with aerodynamics of less than 2.5 (PM2.5) and ozone are the two main air pollutants in China, which seriously endanger human health. To estimate the adverse impacts of PM2.5 and ozone on human health during the implementation of air pollution prevention and control actions in Chengdu, both the generalized additive model and the nonlinear distribution lag model of epidemiology were adopted to explore the exposure-response relationship coefficients ß of daily ozone 8h maximum concentration average (O3-8h), as well as that of PM2.5 on disease deaths in Chengdu from 2014 to 2016. On this basis, the environmental risk model and environmental value assessment model were both adopted to evaluate the health effects and health benefits in Chengdu from 2016 to 2020, respectively, with the assumption that PM2.5 and O3-8h concentration were reduced to specified air pollution control limits (35 µg·m-3 and 70 µg·m-3, respectively). The results showed 1 the annual concentration of PM2.5 presented gradually decreasing trends in Chengdu from 2016 to 2020. Specifically, ρ(PM2.5) from 63 µg·m-3 in 2016 decreased to 40.92 µg·m-3 in 2020. The average annual decline rate was approximately 9.8%. In contrast, the annual concentration of O3-8h from 155 µg·m-3 in 2016 increased to 169 µg·m-3 in 2020, and the increasing rate was approximately 2.4%. 2 Both PM2.5 and O3-8h had lag effects on three types of disease deaths. Under the maximum lag effect, the corresponding exposure-response relationship coefficients ß of PM2.5were 0.0003600, 0.0005001, and 0.0009237 for all-cause, cardiovascular, and respiratory premature deaths, respectively, whereas the corresponding ß of O3-8h were 0.0003103, 0.0006726, and 0.0007002, respectively. 3 If ρ(PM2.5) was reduced to the national secondary standard limit (35 µg·m-3), the corresponding number of health beneficiaries and economic benefits declined yearly. Specifically, the health beneficiary number of all-cause, cardiovascular, and respiratory disease deaths were reduced from 1128, 416, and 328 in 2016 to 229, 96, and 54 in 2020, respectively. There were a total number of 3314 avoidable premature deaths for all-cause diseases during the five years, resulting in a total health economic benefit of 7.66 billion yuan. 4 If we assume that ρ(O3-8h) was reduced to the concentration limit specified by the World Health Organization (70 µg·m-3), the corresponding number of health beneficiaries and economic benefits were increasing yearly. Specifically, the health beneficiaries' numbers of all-cause, cardiovascular, and respiratory disease deaths rose from 1919, 779, and 606 in 2016 to 2429, 1157, and 635 in 2020, respectively. The annual average growth rates of avoidable all-cause and cardiovascular mortality were 6.85% and 10.72%, respectively, which was higher than the annual average rise rate of ρ(O3-8h). There were 10790 total avoidable deaths from all-cause diseases during the five years, resulting in a total health economic benefit of 26.62 billion yuan. These findings indicate that PM2.5 pollution in Chengdu had been well controlled, whereas O3 pollution had become more severe and had become another key air pollutant threatening human health. Therefore, the synchronous control of PM2.5 and ozone should be implemented in the future.

Identification of a Rare BAIAP2-ROS1 Fusion and Clinical Benefit of Crizotinib in the Treatment of Advanced Lung Adenocarcinoma: A Case Report.

Objective: Previous studies have shown that fusion partners have a potential role in influencing different tumorigenic abilities of ROS1 fusion variants, as well as potential differential responses to crizotinib. Therefore, it is important to accurately identify the type of ROS1 rearrangement in NSCLC for clinical treatment selection. Materials and Methods: Deep-coverage targeting solid tumor 31 cancer-related genes panel was used to capture DNA-based NGS information to detect gene fusion. RNA fusion panel based on hybrid capture sequencing was performed to verify gene fusions from total RNA which isolated from formalin fixed paraffin-embedded (FFPE) tissue blocks. Results: Using DNA-targeted NGS method, we identified a novel BAIAP2-ROS1 fusion in a 71-year-old non-smoking female patient with stage IVB lung adenocarcinoma. Rearrangement consisted of BAIAP2 in exon1-exon13 of chr17: q23 and ROS1 in exon35-exon43 of chr6: q22, which were further confirmed by RNA-based NGS methodology. A complete kinase domain in ROS1 fusion was preserved. The patient subsequently received crizotinib and showed significant tumor reduction until 17 months, who got benefit from targeted therapy. Conclusion: This study discovered a novel BAIAP2-ROS1 rearrangement; it provides more knowledge of ROS1 fusion in clinical personalized treatment. The good response to crizotinib therapy emphasizes the importance of DNA-based and RNA-based NGS in rare fusion identification in clinical practice.

Modification effects of ambient temperature on ozone-mortality relationships in Chengdu, China.

A multitude of epidemiological studies have demonstrated that both ambient temperatures and air pollution are closely related to health outcomes. However, whether temperature has modification effects on the association between ozone and health outcomes is still debated. In this study, three parallel time-series Poisson generalized additive models (GAMs) were used to examine the effects of modifying ambient temperatures on the association between ozone and mortality (including non-accidental, respiratory, and cardiovascular mortality) in Chengdu, China, from 2014 to 2016. The results confirmed that the ambient high temperatures strongly amplified the adverse effects of ozone on human mortality; specifically, the ozone effects were most pronounced at > 28 °C. Without temperature stratification conditions, a 10-µg/m3 increase in the maximum 8-h average ozone (O3-8hmax) level at lag01 was associated with increases of 0.40% (95% confidence interval [CI] 0.15%, 0.65%), 0.61% (95% CI 0.27%, 0.95%), and 0.69% (95% CI 0.34%, 1.04%) in non-accidental, respiratory, and cardiovascular mortality, respectively. On days during which the temperature exceeded 28 °C, a 10-µg/m3 increase in O3-8hmax led to increases of 2.22% (95% CI 1.21%, 3.23%), 2.67% (95% CI 0.57%, 4.76%), and 4.13% (95% CI 2.34%, 5.92%) in non-accidental, respiratory, and cardiovascular mortality, respectively. Our findings validated that high temperature could further aggravate the health risks of O3-8hmax; thus, mitigating ozone exposure will be brought into the limelight especially under the context of changing climate.

A scalable platform to discover antimicrobials of ribosomal origin.

Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a promising source of new antimicrobials in the face of rising antibiotic resistance. Here, we report a scalable platform that combines high-throughput bioinformatics with automated biosynthetic gene cluster refactoring for rapid evaluation of uncharacterized gene clusters. As a proof of concept, 96 RiPP gene clusters that originate from diverse bacterial phyla involving 383 biosynthetic genes are refactored in a high-throughput manner using a biological foundry with a success rate of 86%. Heterologous expression of all successfully refactored gene clusters in Escherichia coli enables the discovery of 30 compounds covering six RiPP classes: lanthipeptides, lasso peptides, graspetides, glycocins, linear azol(in)e-containing peptides, and thioamitides. A subset of the discovered lanthipeptides exhibit antibiotic activity, with one class II lanthipeptide showing low µM activity against Klebsiella pneumoniae, an ESKAPE pathogen. Overall, this work provides a robust platform for rapidly discovering RiPPs.

Anti-inflammatory effects of interleukin-23 receptor cytokine-binding homology region rebalance T cell distribution in rodent collagen-induced arthritis.

IL-23 is an important cytokine to regulate Th17 cell differentiation and promote the proliferation of inflammatory cells in Th17-mediated autoimmune diseases. The collagen-induced arthritis (CIA) in rat is a model of rheumatoid arthritis characterized by pronounced inflammatory auto-responses from B and T cells, especially Th17 cells in lesions. In the present study, we used rhIL23R-CHR to block the IL-23 signaling pathway to probe the importance of IL-23 in misbalancing the ratio of Th17/Th9/Treg cells in CIA rats. After treatments with rhIL23R-CHR, the CIA rats showed a significant decrease of secretions of IL-17 and IL-9, whereas FoxP3 was activated in the process, indicating that IL-23 can manipulate the balance of Th17/Th9/Treg cells. Similar to the animal model, IL-23 also possessed remarkable proinflammatory effects on human fibroblast-like synoviocyte cells (HFLS), showing synergetic outcomes with TNF-α. Together, IL-23 could act as a modulator to imbalance the ratio of Th17/Th9/Treg cells, and rhIL23R-CHR could serve as a potential therapeutic agent for RA patients.