Automatic Microfluidic Harmonized RAA-CRISPR Diagnostic System for Rapid and Accurate Identification of Bacterial Respiratory Tract Infections.

Respiratory tract infections (RTIs) pose a grave threat to human health, with bacterial pathogens being the primary culprits behind severe illness and mortality. In response to the pressing issue, we developed a centrifugal microfluidic chip integrated with a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats (CRISPR) system to achieve rapid detection of respiratory pathogens. The limitations of conventional two-step CRISPR-mediated systems were effectively addressed by employing the all-in-one RAA-CRISPR detection method, thereby enhancing the accuracy and sensitivity of bacterial detection. Moreover, the integration of a centrifugal microfluidic chip led to reduced sample consumption and significantly improved the detection throughput, enabling the simultaneous detection of multiple respiratory pathogens. Furthermore, the incorporation of Chelex-100 in the sample pretreatment enabled a sample-to-answer capability. This pivotal addition facilitated the deployment of the system in real clinical sample testing, enabling the accurate detection of 12 common respiratory bacteria within a set of 60 clinical samples. The system offers rapid and reliable results that are crucial for clinical diagnosis, enabling healthcare professionals to administer timely and accurate treatment interventions to patients.

MOFs@POMs-derived bimetallic oxide Fe2(MoO4)3 nanoparticles for sensitive colorimetric detection of salicylic acid in aspirin.

A colorimetric sensing method for salicylic acid (SA) was developed by designing and fabricating bimetallic oxide nanozymes. Firstly, by calcinating MIL-100(Fe)@PMo12 (MOFs@POMs) at different temperature, Fe2(MoO4)3-Ts (T = 400℃, 500℃, 600℃, 700℃) nanoparticles (NPs) were successfully prepared. Secondly, by evaluating the peroxidase-like activities, Fe2(MoO4)3-600 NPs shows the best peroxidase-like activity attributed to the Fenton-like effect and the synergistic coupling interaction between Mo and Fe. Finally, based on the specific complexation between SA and Fe3+, a sensitive colorimetric sensor for SA was established, which exhibits superior selectivity and interference with a detection limit of 0.11 µM and a linear range of 10 to 100 µM, the lowest LOD for SA to date, to the best of our knowledge.

Immunomagnetic Separation Combined with RCA-CRISPR/Cas12a for the Detection of Salmonella typhimurium on a Figure-Actuated Microfluidic Biosensor.

A figure-actuated microfluidic biosensor was developed for the rapid and sensitive detection of Salmonella typhimurium using immunomagnetic separation to separate target bacteria and rolling circle amplification (RCA) combined with CRISPR/Cas12a to amplify the detection signal. The magnetic nanoparticles (MNPs) modified with the capture antibodies (MNPs@Ab1) and RCA primer linked with recognized antibodies (primer@Ab2) were first used to react with S. typhimurium, resulting in the formation of MNPs@Ab1-S. typhimurium-primer@Ab2 complexes. Then, the RCA and CRISPR/Cas12a reagents were successively pumped into the chamber and incubated at the appropriate conditions. With the help of a 3D-printed signal detector, the fluorescence signal was collected and analyzed using the smartphone APP for the determination of bacterial concentration. This biosensor exhibited a wide linear range for the detection of S. typhimurium with a low limit of detection of 1.93 × 102 CFU/mL and a mean recovery of about 106% in the spiked milk sample.

Increasing the peroxidase-like activity of the MIL-100(Fe) nanozyme by encapsulating Keggin-type 12-phosphomolybdate and covering three-dimensional graphene.

To improve the peroxidase-like activities of metal-organic frameworks (MOFs) as nanozymes, a ternary MIL-100(Fe)@PMo12@3DGO nanocomposite was designed and fabricated by encapsulating Keggin-type H3PMo12O40 (PMo12) with fast and reversible multi-electron redox processes and an electron-rich structure into MIL-100(Fe), then being covered by three-dimensional graphene (3DGO) with higher conductivity, larger surface area, higher porosity, and better chemical stability. As a consequence, the as-prepared MIL-100(Fe)@PMo12@3DGO nanocomposite exhibits excellent peroxidase-like activities, namely, the lowest limit of detection (0.14 µM) in the range of 1-100 µM for glucose to date, to the best of our knowledge, attributed to the individual and synergistic effects of H3PMo12O40, 3DGO and MIL-100(Fe).

Analysis of the Renal Protection and Antioxidative Stress Effects of Panax notoginseng Saponins in Diabetic Nephropathy Mice.

Objective: Diabetic nephropathy (DN), a diabetes-induced chronic complication, is the major trigger of end-stage renal disease. As the main active ingredient of Panax notoginseng (PNG), Panax notoginseng saponins (PNS) are crucial in treating renal diseases. This study is aimed at investigating the role played by PNS in renal protection and antioxidative stress (OS) in DN mice. Methods: A DN mouse model was constructed, and then low, medium, and high doses of PNS were used to intervene the model group mice. Eight weeks after intervention, the 24 h urine protein (UPro) and urinary albumin (UAlb) were quantitatively examined, and the related blood biochemical indices were measured. HE and PAS staining were performed for pathological changes of renal tissue. ELISA and western blotting were carried out to quantify the levels of OS indexes and inflammatory factors (IFs) in mouse kidney tissues and the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1), respectively. Results: The weight of DN mice decreased first compared with control animals and then gradually increased after different doses of PNS treatment. Besides, DN mice presented elevated urine volume, UPro, and UAlb, all of which were reversed by PNS intervention. SOD activity and GSH content in renal tissues of the model group mice decreased markedly versus the control group, and MDA, CRP, IL-6, and TGF-ß1 contents elevated statistically, while different doses of PNS effectively reduced the OS injury and IFs in mice. Compared with the model group, PNS dose-dependently increased Nrf2 and HO-1 levels in DN mice. Conclusions: PNS is protective of HFF + STZ-induced DN mice against kidney tissue damage and can reduce the excretion of UPro and relieve the OS state of mice, possibly by activating Nrf2/HO-1 axis to play an antioxidant and anti-inflammatory role.

MHDMF: Prediction of miRNA-disease associations based on Deep Matrix Factorization with Multi-source Graph Convolutional Network.

A growing number of works have proved that microRNAs (miRNAs) are a crucial biomarker in diverse bioprocesses affecting various diseases. As a good complement to high-cost wet experiment-based methods, numerous computational prediction methods have sprung up. However, there are still challenges that exist in making effective use of high false-negative associations and multi-source information for finding the potential associations. In this work, we develop an end-to-end computational framework, called MHDMF, which integrates the multi-source information on a heterogeneous network to discover latent disease-miRNA associations. Since high false-negative exist in the miRNA-disease associations, MHDMF utilizes the multi-source Graph Convolutional Network (GCN) to correct the false-negative association by reformulating the miRNA-disease association score matrix. The score matrix reformulation is based on different similarity profiles and known associations between miRNAs, genes, and diseases. Then, MHDMF employs Deep Matrix Factorization (DMF) to predict the miRNA-disease associations based on reformulated miRNA-disease association score matrix. The experimental results show that the proposed framework outperforms highly related comparison methods by a large margin on tasks of miRNA-disease association prediction. Furthermore, case studies suggest that MHDMF could be a convenient and efficient tool and may supply a new way to think about miRNA-disease association prediction.

The experimental study of fluorine-leaching ability of granite with different solutions: A new insight into the dynamic of groundwater fluorine levels along coastal zones.

Drinking-water fluorosis is universal along coastal zones, and the seawater or brine water intrusion is occasionally supposed to enrich groundwater fluorine levels. However, there is no conclusive proof, and the laws and mechanisms remain ambiguous. Granite, the common fluorine-bearing rock, is selected and experimented upon to reveal the characteristics and laws of fluorine's leaching ability during the intrusion of seawater. The fluorine-leaching ability increases with the increasing ratios of seawater or brine water, the increasing levels of NaCl or NaHCO3, and the decreasing levels of CaCl2. Such results directly confirm that seawater or brine water intrusion, as well as the conditions of higher Na+, HCO3- and lower Ca2+, promotes fluorine-leaching ability from granite. The intensities of SiOSi, SiOFe, SiOAl bonds decrease but those of OH bonds increase with a higher ratio of seawater or brine water, the higher levels of NaCl or NaHCO3, and the lower levels of CaCl2. This indicates the more silicate dissolution and stronger OH-F exchange evoked by seawater or brine water intrusion are responsible for the higher fluorine-leaching from granite. Therefore, the process of seawater or brine water intrusion should be important for the groundwater enrichment dynamics along coastal zones.

Effects of gender affirming hormone therapy on body mass index in transgender individuals: A longitudinal cohort study.

INTRODUCTION: Many transgender people take hormone therapy to affirm their gender identity. One potential long-term consequence of gender affirming hormone therapy is increased body mass index (BMI), which may be associated with metabolic syndrome, cardiovascular disease and higher mortality. Only a few published studies explored changes in BMI in transgender people taking gender affirming hormone therapy (GAHT). OBJECTIVE: To examine the changes in BMI longitudinally in response to GAHT in transgender women and men. METHODS: We conducted a retrospective cohort study of transgender individuals who received GAHT from the endocrinology clinic between January 1, 2000 and September 6, 2018. Subjects who sought GAHT were included if they had two separate measurements of BMI and were excluded if they had a BMI greater than 35 kg/m2 or were missing demographic data at entry. We used a linear mixed model to analyze the longitudinal change in BMI. RESULTS: There were a total of 227 subjects included in this cohort. Among subjects already on GAHT, transgender women were receiving GAHT longer than transgender men (6.59 ± 9.35 vs 3.67 ± 3.43 years, p-value = 0.04). Over the period of 7 years, there was a significant increase in BMI in transwomen who newly initiated GAHT (p-value 0.004). There were no changes in BMI in transgender men and women already on GAHT or in transgender men who newly initiated GAHT in the study. CONCLUSION: We conclude that BMI significantly increases in transwomen but not in transmen after initiation of GAHT in a single center based in the United States. In transwomen and transmen, BMI appears to be stable following 3 to 6 years of GAHT. Future investigations should examine the causes for increased BMI in transgender women including type of GAHT, diet and lifestyle, and association with risk of metabolic syndrome and cardiovascular disease.

Prediction of acute GVHD and relapse by metabolic biomarkers after allogeneic hematopoietic stem cell transplantation.

BACKGROUND: There are very few studies investigating metabolic biomarkers to predict acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). Metabolic models can provide a framework for analyzing the information-rich omics data sets in this setting. METHODS: Four hundred and fifty-six samples from one hundred and fourteen consecutive patients who underwent HSCT from January 2012 to May 2014 were collected for this study. The changes in serum metabolite levels were investigated using a gas chromatography-mass spectrometry-based metabolomics approach and underwent statistical analysis. RESULTS: Significant metabolic changes were observed on day 7. The stearic acid/palmitic acid (SA/PA) ratio was effective in the diagnosis of grade II-IV aGVHD. Multivariate analysis showed that patients with high SA/PA ratios on day 7 after HSCT were less likely to develop II-IV aGVHD than patients with low SA/PA ratios (odds ratio [OR] = 0.06, 95% CI 0.02-0.18, P < 0.001). After the adjustment for clinical characteristics, the SA/PA ratio had no significant effect on overall survival (hazard ratio [HR] = 1.95, 95% CI 0.92-4.14, P = 0.08), and patients in the high SA/PA ratio group were significantly more likely to relapse than those in the low ratio group (HR = 2.26, 95% CI 1.04-4.91, P = 0.04). CONCLUSION: Our findings suggest that the SA/PA ratio on day 7 after HSCT is an excellent biomarker to predict both aGVHD and relapse. The serum SA/PA ratio measured on day 7 after transplantation may improve risk stratification for aGVHD and relapse after allogeneic stem cell transplantation. FUNDING: National Natural Science Foundation of China (81470346, 81773361), Priority Academic Program Development of Jiangsu Higher Education Institutions, Jiangsu Natural Science Foundation (BK20161204), Innovation Capability Development Project of Jiangsu Province (BM2015004), Jiangsu Medical Junior Talent Person award (QNRC2016707), and NIH (AI129582 and NS106170).

Characterizing the Tumor Suppressor Role of CEACAM1 in Multiple Myeloma.

BACKGROUND/AIMS: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), also known as CD66a, is a member of the immunoglobulin (Ig) superfamily that belongs to the carcinoembryonic antigen (CEA) family which plays a dual role in cancer. Previous studies showed high expression of CEACAM1 in multiple myeloma (MM). The aim of this study was to investigate the biological consequences of CEACAM1 overexpression in MM. METHODS: pEGFP-N1-CEACAM1 and pcDNA3.1-CEACAM1 expression plasmids were transfected into U-266 and RPMI8266 cell lines . Effect of CEACAM1 overexpression on the proliferation of two cell lines were tested by the CCK8 assay. Cell cycle and Apoptotic changes after CEACAM1 transfection were examined with AnnexinV-FITC/PI by flow cytometry. Hochest staining assay was used to confirm the apoptotic changes. Caspase-3 activity was examined by Western blotting. The cell invasion and migration activity change after CEACAM1 transfection were performed by well chamber assays and a wound healing, respectively. MMP-2 and MMP-9 proteins expression were detected by Western blotting. Flow cytometry immunophenotyping was be evaluated on myeloma cells from bone marrow taken from 50 patients with symptomatic MM newly diagnosed. The correlations between CEACAM1 expression levels and the clinical features across all groups were investigated. RESULTS: CEACAM1 overexpression significantly suppressed MM cell proliferation, induced cell apoptosis, and inhibited cell invasion and migration possibly through activation of caspase-3 and downregulation of MMP-2 and MMP-9. CEACAM1 expression in patients with DS stage I was more frequent (61.5%) than those with DS stage II (21.1%) or III (22.2%). Furthermore, patients with ß2-microglobulin levels equal to or less than 3.5 mg/L had higher CEACAM1 expression than those with ß2-microglobulin levels greater than 3.5 mg/L. CONCLUSION: Our findings suggest that CEACAM1 may act as a tumor suppressor in MM.

Oxidation of HRas cysteine thiols by metabolic stress prevents palmitoylation in vivo and contributes to endothelial cell apoptosis.

Here we demonstrate a new paradigm in redox signaling, whereby oxidants resulting from metabolic stress directly alter protein palmitoylation by oxidizing reactive cysteine thiolates. In mice fed a high-fat, high-sucrose diet and in cultured endothelial cells (ECs) treated with high palmitate and high glucose (HPHG), there was decreased HRas palmitoylation on Cys181/184 (61±24% decrease for cardiac tissue and 38±7.0% in ECs). This was due to oxidation of Cys181/184, detected using matrix-assisted laser desorption/ionization time of flight (MALDI TOF)-TOF. Decrease in HRas palmitoylation affected its compartmentalization and Ras binding domain binding activity, with a shift from plasma membrane tethering to Golgi localization. Loss of plasma membrane-bound HRas decreased growth factor-stimulated ERK phosphorylation (84±8.6% decrease) and increased apoptotic signaling (24±6.5-fold increase) after HPHG treatment that was prevented by overexpressing wild-type but not C181/184S HRas. The essential role of HRas in metabolic stress was made evident by the similar effects of expressing an inactive dominant negative N17-HRas or a MEK inhibitor. Furthermore, the relevance of thiol oxidation was demonstrated by overexpressing manganese superoxide dismutase, which improved HRas palmitoylation and ERK phosphorylation, while lessening apoptosis in HPHG treated ECs.