Real-time, random-access organ screening for carbapenem-resistant organisms (CRO) reduces CRO-associated, donor-derived infection mortality in lung transplant recipients.

PURPOSE: Donor-derived infection (DDI) has become an important factor affecting the prognosis of lung transplantation patients. The risks versus benefits of using donor organs infected with multidrug-resistant organisms (MDRO), especially carbapenem-resistant organisms (CRO), are frequently debated. Traditional microbial culture and antimicrobial susceptibility testing at present fail to meet the needs of quick CRO determination for donor lungs before acquisition. In this study, we explored a novel screening method by using Xpert® Carba-R assay for CRO in donor lungs in a real-time manner to reduce CRO-associated DDI mortality. METHODS: This study was registered on chictr.org.cn (ChiCTR2100053687) on November 2021. In the Xpert Carba-R screening group, donor lungs were screened for CRO infection by the Xpert Carba-R test on bronchoalveolar fluid (BALF) before acquisition. If the result was negative, donor lung acquisition and subsequent lung transplantation were performed. In the thirty-five potential donors, nine (25.71%) with positive Xpert Carba-R results in BALF were declined for lung transplantation. Twenty-six recipients and the matching CRO-negative donor lungs (74.29%) were included in the Xpert Carba-R screening group. In the control group, nineteen recipients underwent lung transplants without Xpert Carba-R screening. The incidence and mortality of CRO-associated DDI were collected and contrasted between the two groups. RESULTS: Multivariate analysis showed that CRO-related death due to DDI within 60 days was significantly lower in the Xpert Carba-R screening group than that in the control group (OR = 0.05, 95% CI 0.003-0.74, p = 0.03). CONCLUSION: Real-time CRO screening of donor lungs before transplantation at the point of care by the Xpert Carba-R helps clinicians formulate lung transplantation strategies quickly and reduces the risk of subsequent CRO infection improving the prognosis of lung transplantation.

Pharmacokinetic Evaluation of Tacrolimus in Chinese Adult Patients during the Early Stages Post-Lung Transplantation.

BACKGROUND: Although tacrolimus has been widely used in patients undergoing lung transplantation, few studies have reported the pharmacokinetics of tacrolimus in Chinese patients after lung transplantation. Thus, we aimed to investigate the pharmacokinetics and influential factors in this patient cohort in the early stage after lung transplantation. METHODS: We enrolled 14 adult lung transplant recipients who were treated with tacrolimus and then intensively collected blood samples within a 12-h dosing interval. The pharmacokinetic parameters of tacrolimus were calculated using non-compartmental analysis, and the influence of pathophysiological characteristics and CYP3A5*3 and CYP3A4*1G genotypes on the pharmacokinetics of tacrolimus was assessed. Using linear regression analysis, we investigated the correlation between tacrolimus concentration at different sampling points and measured the area under the time-concentration curve (AUC0-12h). RESULTS: Geometric mean of apparent clearance (CL/F) was 18.13 ± 1.65 L/h in non-CYP3A5*3/*3 carriers, five times higher than that in CYP3A5*3/*3 carriers (p < 0.001). Furthermore, the tacrolimus concentration 4 h after administration had the strongest correlation with AUC0-12h (R2 = 0.979). CONCLUSION: The pharmacokinetics of tacrolimus varied largely between patients during the early stage post-transplantation, which could be partially explained by CYP3A5*3 genetic polymorphisms.

Extraction optimization and screening of antioxidant peptides from grass carp meat and synergistic-antagonistic effect.

Grass carp (Ctenopharyngodon idellus) is one of the three most cultivated freshwater fish around the world, but it is mainly consumed afresh, so only a small part of them are processed into salted fish or snack food. This research was performed to prepare and screen antioxidant peptides from grass carp muscle to promote its high-value utilization. The parameters of double-enzyme two-step hydrolysis were optimized, the peptides with the highest ABTS.+ scavenging ability were enriched and identified by Sephadex G-25 and LC-Q-Orbitrap-MS/MS. The synergistic-antagonistic effect among identified peptides was also investigated. The optimized conditions were hydrolyzed with protamex (10,000 U/g) at pH 8.0, 50°C for 3 h, followed by hydrolysis with alcalase (6,000 U/g) at pH 9.0, 50 °C for 2 h, and the protein-liquid ratio was 4%. The hydrolysates were further fractionated to obtain five fractions, in which fraction 3 (F3) exhibited the strongest ABTS.+ and O 2 · - scavenging ability with the IC50 values of 0.11 and 0.47 mg/ml, respectively. Twelve novel antioxidant peptides were identified, in which VAGW possessed the highest activity (139.77 µmol GSH/g). Significantly synergistic effects were observed on the two and three peptides' combination among VAGW, APPAMW, LFGY, FYYGK, and LLLYK, while the C-terminal tryptophan (Trp) played an important role in the synergism. This study found that grass carp muscle hydrolysates can be potential natural antioxidants in functional products. The synergistic effects among peptides may provide a perspective for the combined application of peptides.

Glycation of ß-lactoglobulin combined by sonication pretreatment reduce its allergenic potential.

ß-lactoglobulin (ß-Lg) was treated through different ultrasonic power and subsequently glycated with galactose to investigate its structural changes and immunological properties, and then evaluated by high-resolution mass spectrometry, enzyme-linked immunosorbent assay and basophil histamine release test. Ultrasonication combined with glycation (UCG) modification significantly reduced the IgE/IgG-binding capacity, and the release of ß-hexosaminidase, histamine and interleukin-6, accompanied with changes in the secondary and tertiary structures. The decrease in the allergenicity of ß-Lg depended not only on the glycation of K47, 60, 83, 91 and 135 within the linear epitopes, but also on the denaturation of conformational epitopes, which was supported by the glycation-induced alterations of the secondary and tertiary structures. This study confirmed that UCG modification is a promising method for decreasing the allergenic potential of allergic proteins, which is likely to develop a practical technology to produce hypo-allergenic milk.

Antioxidant, metabolic enzymes inhibitory ability of Torreya grandis kernels, and phytochemical profiling identified by HPLC-QTOF-MS/MS.

In this study, the antioxidant activities, α-glucosidase and tyrosinase inhibitory ability of Torreya grandis kernels (TGK) were performed. Samples were extracted with various polarity of ethanol, and the major phytochemical profile was characterized. The results showed that 70% of ethanol extract gave the richest phenolics and flavonoids. The strongest DPPH· and ABTS·+ scavenging ability, as well as the best inhibition on tyrosinase and α-glucosidase was also detected on 70% of ethanol extract. Among the fractions of 70% of ethanol extract, the ethyl acetate fraction (EAF) owned the highest phenolics, flavonoids, and the best DPPH· and ABTS·+ scavenging ability, and tyrosinase inhibition. Unexpectedly, the dichloromethane fraction possessed the strongest inhibition on α-glucosidase, which was much greater than that of acarbose. HPLC-QTOF-MS/MS analysis result to the characterization of 19 compounds from EAF. The results implied that TGK can be a potential source of natural antioxidants, α-glucosidase and tyrosinase inhibitors. Practical applications The kernels of T. grandis are one of the precious nuts in the world, and the extracts were advertised to show a variety of biological activities and pharmacological effects. However, researches on the phytochemical constituents and bioactivities are fewer. In this study, TGK was found to show good potency in antioxidant, α-glucosidase and tyrosinase inhibitory activities. The 70% ethanol is the best solvent for extracting above mentioned active components, and ethyl acetate can be the suitable enriching solvent. In addition, the predominant phytochemical compounds in EAF were characterized. Therefore, this research can help to the performance of further research and application of TGK in functional products.

Up-regulation of growth factor independence 1 in endotoxin tolerant macrophages with low secretion of TNF-alpha and IL-6.

OBJECTIVE: Endotoxin tolerance refers to a low response to lipopolysaccharide (LPS). We hypothesized that growth factor independence 1 (Gfi1) involves in the endotoxin tolerance in macrophages. METHODS: Endotoxin tolerance was induced in the RAW264.7 cell line and thioglycolate-elicited murine peritoneal macrophages by incubation with 100 ng/ml LPS for 20 h. Macrophages without the pretreatment were set as control. Both endotoxin tolerant and control cells were then stimulated with 1,000 ng/ml LPS for indicated period of incubation. Gfi1 mRNA expression and protein production were investigated by real-time PCR and Western blotting, respectively. ELISA was performed to quantify the secretion of TNF-alpha and IL-6. RESULT: Compared with non-endotoxin tolerant macrophages, endotoxin tolerant cells secreted a lower amount of TNF-alpha and IL-6. The mRNA expression of Gfi1 in endotoxin tolerant macrophages was higher than that of control in both RAW264.7 cells and thioglycolate-elicited murine peritoneal macrophages. The protein production was accordingly up-regulated in endotoxin tolerant RAW264.7 cells. CONCLUSION: In in vitro endotoxin tolerant macrophages, the expression of Gfi1 mRNA and protein were up-regulated after high dose LPS stimulation, accompanied with a blunted TNF-alpha and IL-6 secretion. Gfi1 might participate in the mechanism of endotoxin tolerance.