DataSheet_1_Optimal selection of specimens for metagenomic next-generation sequencing in diagnosing periprosthetic joint infections.docx

ObjectiveThis study aimed to assess the diagnostic value of metagenomic next-generation sequencing (mNGS) across synovial fluid, prosthetic sonicate fluid, and periprosthetic tissues among patients with periprosthetic joint infection (PJI), intending to optimize specimen selection for mNGS in these patients.MethodsThis prospective study involved 61 patients undergoing revision arthroplasty between September 2021 and September 2022 at the First Affiliated Hospital of Zhengzhou University. Among them, 43 cases were diagnosed as PJI, and 18 as aseptic loosening (AL) based on the American Musculoskeletal Infection Society (MSIS) criteria. Preoperative or intraoperative synovial fluid, periprosthetic tissues, and prosthetic sonicate fluid were collected, each divided into two portions for mNGS and culture. Comparative analyses were conducted between the microbiological results and diagnostic efficacy derived from mNGS and culture tests. Furthermore, the variability in mNGS diagnostic efficacy for PJI across different specimen types was assessed.ResultsThe sensitivity and specificity of mNGS diagnosis was 93% and 94.4% for all types of PJI specimens; the sensitivity and specificity of culture diagnosis was 72.1% and 100%, respectively. The diagnostic sensitivity of mNGS was significantly higher than that of culture (X2 = 6.541, P=0.011), with no statistically significant difference in specificity (X2 = 1.029, P=0.310). The sensitivity of the synovial fluid was 83.7% and the specificity was 94.4%; the sensitivity of the prosthetic sonicate fluid was 90.7% and the specificity was 94.4%; and the sensitivity of the periprosthetic tissue was 81.4% and the specificity was 100%. Notably, the mNGS of prosthetic sonicate fluid displayed a superior pathogen detection rate compared to other specimen types.ConclusionmNGS can function as a precise diagnostic tool for identifying pathogens in PJI patients using three types of specimens. Due to its superior ability in pathogen identification, prosthetic sonicate fluid can replace synovial fluid and periprosthetic tissue as the optimal sample choice for mNGS.</p

Principal coordinate analysis of 11 PWWTP and STP sludge samples based on the percentages of ARG subtypes.

Principal coordinate analysis of 11 PWWTP and STP sludge samples based on the percentages of ARG subtypes.</p

In Situ Visualization of Reversible Diels–Alder Reactions with Self-Reporting Aggregation-Induced Emission Luminogens

The dynamic reversible Diels–Alder (DA) reactions play essential roles in both academic and applied fields. Currently, in situ visualization and direct monitoring of the formation and cleavage of covalent bonds in DA reactions are hampered by finite compatibility and expensive precise instruments, especially limited in solid reactions. We herein report a fluorescence system capable of in situ visualization by naked eyes and monitoring DA/retro-DA reactions. With the fluorescence quenching effect, the synthesized TPEMI could work as an innovative self-indicator for both DA termination and retro-DA occurrence. The fluorescence increases during DA reactions, and the mechanism is investigated to establish qualitative and quantitative relations. Besides rapid screening of reaction conditions and monitoring of DA exchange processes, the TPEMI fluorescence system can visualize heterogeneous and solid-state reactions with the AIE character. The TPEMI platform is expected to offer novel insights into reversible DA processes and dynamic covalent chemistry

Percentage of the predominant ARG subtypes (≥1% in at least one sludge sample) in PWWTP and STP sludge.

<p>Percentage of the predominant ARG subtypes (≥1% in at least one sludge sample) in PWWTP and STP sludge.</p

The abundance and diversity of MGEs in PWWTP sludge.

<p>The abundance and diversity of MGEs in PWWTP sludge.</p

Redundancy analysis of the correlation between the percentages of 36 dominant genera with the percentages of eight ARG types in PWWTP sludge.

<p>Arrows represent the seven genera significantly correlated with ARGs distribution (<i>p</i><0.05).</p

Abundance of ARGs in the PWWTP and STP sludge samples.

<p>(A) Total abundance of ARGs in the PWWTP and STP sludge. (B) Percentage of the ARG types in the PWWTP and STP sludge. (C) Percentage of resistance mechanisms in the PWWTP and STP sludge.</p

Bacterial community in the PWWTP sludge samples.

<p>(A) Percentages of different phyla in the PWWTP sludge. “Others (1%)” refers to the phyla with abundance <1% in all samples. (B) Percentages of top 10 genera in the PWWTP sludge samples. The asterisks indicate the significant difference with <i>p</i>-value<0.05 between anaerobic and aerobic sludge samples.</p

Treatment processes in the two PWWTPs and the shared ARGs.

<p>(A) The flow charts and sampling sites of the treatment processes in two PWWTPs. The black points in oxidation ditch represent the sampling sites. HA: Hydrolytic Acidification; CASS: Cyclic Activated Sludge System; UBF: Up-flow Blanket Filter; (B) Number of shared ARG subtypes by adjacent treatment systems in two PWWTPs. (C) Percentage of the shared ARGs in the total ARGs.</p