The function of an apple ATP-dependent Phosphofructokinase gene MdPFK5 in regulating salt stress.

Salt stress severely affects the growth and yield of apples (Malus domestica Borkh). Although salt-tolerant genes have been extensively studied, documentation on the role of the ATP-dependent phosphofructokinase gene MdPFK5 in salt stress is limited. This study conducted an evolutionary tree and three-dimensional structure analysis of the PFK gene family in Arabidopsis thaliana and MdPFK (MD01G1037400), revealing a close phylogenetic relationship between MdPFK (MD01G1037400) and AtPFK5. Given the similarity in their protein tertiary structures, MdPFK was designated as MdPFK5, suggesting functional similarities with AtPFK5. Further investigation revealed elevated expression levels of MdPFK5 in apple leaves and flowers, particularly showing significant upregulation 120 days after blooming and differential expression beginning at 3 hours of salt stress. Overexpression of MdPFPK5 conferred salt tolerance in both apple calli and transgenic lines of Arabidopsis thaliana. Moreover, NaCl treatment promoted soluble sugar accumulation in apple calli and transgenic lines of Arabidopsis thaliana overexpressing MdPFK5. This study provides new insights into the salt tolerance function of MdPFK5.

The impact of supplementing vitamin D through different methods on the prognosis of COVID-19 patients: a systematic review and meta-analysis.

Objective: To analyze the impact of different methods of Vitamin D administration on the prognosis of COVID-19 patients. Methods: A comprehensive literature search was conducted across four databases: PubMed, Embase, Web of Science, and Cochrane, up to January 5, 2024. Eligible studies included randomized controlled trials and cohort studies that compared Vitamin D supplementation with control groups in COVID-19 patients. Outcomes of interest were mortality rate, ICU (Intensive Care Unit) admission rate, length of hospital stay, and endotracheal intubation rate. Subgroup analyses were performed based on the dosing regimen (single-dose vs. continuous-dose), total Vitamin D intake within 14 days (≥100,000 IU vs. <100,000 IU), and baseline serum Vitamin D levels (deficient group: 25OHD < 30 ng/mL vs. non-restricted group). A random-effects model was employed for meta-analysis to account for heterogeneity among studies. Results: A total of 21 studies involving 4,553 participants were included. In terms of mortality, Vitamin D supplementation significantly reduced the mortality rate (RR = 0.72, 95% CI: 0.54-0.94, I 2 = 54%, p = 0.02), with continuous dosing being more effective (RR = 0.53, 95% CI: 0.34-0.83, I 2 = 55%, p = 0.006) compared to single-dose (RR = 0.88, 95% CI: 0.69-1.12, I 2 = 21%, p = 0.3), and lower total doses (<100,000 IU) showing greater benefit (RR = 0.30, 95% CI: 0.21-0.44, I 2 = 0%, p < 0.0001). Mortality was significantly reduced in the Vitamin D-deficient group (25OHD < 30 ng/mL) (RR = 0.73, 95% CI: 0.59-0.89, I 2 = 0%, p = 0.002) but not in the non-restricted group. Regarding ICU admission, supplementation reduced ICU admission rates (RR = 0.58, 95% CI: 0.38-0.88, I 2 = 74%, p = 0.01), with continuous dosing (RR = 0.44, 95% CI: 0.22-0.90, I 2 = 74%, p = 0.02) being more effective than single-dose (RR = 0.79, 95% CI: 0.61-1.03, I 2 = 22%, p = 0.08), and lower doses (<100,000 IU) providing more significant reduction (RR = 0.31, 95% CI: 0.21-0.47, I 2 = 0%, p = 0.001). ICU admission rates were significantly reduced in the Vitamin D-deficient group (RR = 0.63, 95% CI: 0.42-0.93, I 2 = 0%, p = 0.02) but not in the non-restricted group (RR = 0.59, 95% CI: 0.32-1.11, I 2 = 86%, p = 0.1). For length of hospital stay, no significant differences were observed between Vitamin D and control groups (MD = -1, 95% CI: -2.16 to 0.16, p = 0.13), and subgroup analyses by dosing regimen, total dose, and baseline Vitamin D levels also showed no significant differences. Similarly, for endotracheal intubation, there was no significant difference in intubation rates between groups (RR = 0.78, 95% CI: 0.56-1.08, p = 0.13), and subgroup analyses confirmed no significant effect of different dosing strategies or baseline Vitamin D status on intubation rates. Conclusion: Vitamin D supplementation improves clinical outcomes in COVID-19 patients by reducing mortality and ICU admission rates, particularly when administered continuously with a total dose of less than 100,000 IU over 14 days, and among those with baseline Vitamin D deficiency (25OHD < 30 ng/mL). However, there were no significant effects on the length of hospital stay or endotracheal intubation rates, regardless of the dosing regimen or baseline Vitamin D levels. These findings emphasize the importance of considering both the total dose over 14 days and baseline Vitamin D status to optimize therapeutic benefits.

Enzymatic Synthesis of Novel Terpenoid Glycoside Derivatives Decorated with N-Acetylglucosamine Catalyzed by UGT74AC1.

The addition of the O-linked N-acetylglucosamine (O-GlcNAc) is a significant modification for active molecules, such as proteins, carbohydrates, and natural products. However, the synthesis of terpenoid glycoside derivatives decorated with GlcNAc remains a challenging task due to the absence of glycosyltransferases, key enzymes for catalyzing the transfer of GlcNAc to terpenoids. In this study, we demonstrated that the enzyme mutant UGT74AC1T79Y/L48M/R28H/L109I/S15A/M76L/H47R efficiently transferred GlcNAc from uridine diphosphate (UDP)-GlcNAc to a variety of terpenoids. This powerful enzyme was employed to synthesize GlcNAc-decorated derivatives of terpenoids, including mogrol, steviol, andrographolide, protopanaxadiol, glycyrrhetinic acid, ursolic acid, and betulinic acid for the first time. To unravel the mechanism of UDP-GlcNAc recognition, we determined the X-ray crystal structure of the inactivated mutant UGT74AC1His18A/Asp111A in complex with UDP-GlcNAc at a resolution of 1.66 Å. Through molecular dynamic simulation and activity analysis, we revealed the molecular mechanism and catalytically important amino acids directly involved in the recognition of UDP-GlcNAc. Overall, this study not only provided a potent biocatalyst capable of glycodiversifying natural products but also elucidated the structural basis for UDP-GlcNAc recognition by glycosyltransferases.

Construction of 5-Amino-1,2-Selenazole Scaffolds through N-Selenocyanation/Cyclization of Enaminones Using KSeCN.

A metal-free and mild approach for constructing 5-amino-1,2-selenazole skeletons by NBS/KSeCN-mediated N-selenocyanation and nucleophilic cyclization of ß-enaminones has been developed. Various isoselenazole compounds and the isoselenazolyl derivatives of anti-inflammatory medicines, including isosepac, oxaprozin, and ibuprofen, have been obtained with good yields. This efficient, "one-pot", and atomic economy strategy may represent an alternative route for the construction of a 1,2-selenazole framework via the "+SeCN" pathway and provide new access to heterocycles containing a Se-N bond.

Genetic alteration of SJ293TS cells and modification of serum-free media enhances lentiviral vector production.

Successful cell and gene therapy clinical trials have resulted in the US Food and Drug Administration and European Medicines Agency approving their use for treatment of patients with certain types of cancers and monogenetic diseases. These novel therapies, which rely heavily on lentiviral vectors to deliver therapeutic transgenes to patient cells, have driven additional investigations, increasing demand for both pre-clinical and current Good Manufacturing Practices-grade viral vectors. To better support novel studies by improving current production methods, we report the development of a genetically modified HEK293T-based cell line that is null for expression of both Protein Kinase R and Beta-2 microglobulin and grows in suspension using serum-free media, SJ293TS-DPB. Absence of Protein Kinase R increased anti-sense lentiviral vector titers by more than 7-fold, while absence of Beta-2 microglobulin, a key component of major histocompatibility complex class I molecules, has been reported to reduce the immunogenicity of lentiviral particles. Furthermore, we describe an improved methodology for culturing SJ293TS-DPB that facilitates expansion, reduces handling, and increases titers by 2-fold compared with previous methods. SJ293TS-DPB stably produced lentiviral vectors for over 4 months and generated lentiviral vectors that efficiently transduce healthy human donor T cells and CD34+ hematopoietic stem cells.

Maize Root Exudates Promote Bacillus sp. Za Detoxification of Diphenyl Ether Herbicides by Enhancing Colonization and Biofilm Formation.

Diphenyl ether herbicides are extensively utilized in agricultural systems, but their residues threaten the health of sensitive rotation crops. Functional microbial strains can degrade diphenyl ether herbicides in the rhizosphere of crops, facilitating the restoration of a healthy agricultural environment. However, the interplay between microorganisms and plants in diphenyl ether herbicides degradation remains unclear. Thus, the herbicide-degrading strain Bacillus sp. Za and the sensitive crop, maize, were employed to uncover the interaction mechanism. The degradation of diphenyl ether herbicides by strain Bacillus sp. Za was promoted by root exudates. The strain induced root exudate re-secretion in diphenyl ether herbicide-polluted maize. We further showed that root exudates enhanced the rhizosphere colonization and the biofilm biomass of strain Za, augmenting its capacity to degrade diphenyl ether herbicide. Root exudates regulated gene fliZ, which is pivotal in biofilm formation. Wild-type strain Za significantly reduced herbicide toxicity to maize compared to the ZaΔfliZ mutant. Moreover, root exudates promoted strain Za growth and chemotaxis, which was related to biofilm formation. This mutualistic relationship between the microorganisms and the plants demonstrates the significance of plant-microbe interactions in shaping diphenyl ether herbicide degradation in rhizosphere soils. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 "No Rights Reserved" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2024.

Persistent high sepsis-induced coagulopathy and sequential organ failure assessment scores can predict the 28-day mortality of patients with sepsis: A prospective study.

BACKGROUND: The performance of the sepsis-induced coagulopathy (SIC) and sequential organ failure assessment (SOFA) scores in predicting the prognoses of patients with sepsis has been validated. This study aimed to investigate the time course of SIC and SOFA scores and their association with outcomes in patients with sepsis. METHODS: This prospective study enrolled 209 patients with sepsis admitted to the emergency department. The SIC and SOFA scores of the patients were assessed on days 1, 2, and 4. Patients were categorized into survivor or non-survivor groups based on their 28-day survival. We conducted a generalized estimating equation analysis to evaluate the time course of SIC and SOFA scores and the corresponding differences between the two groups. The predictive value of SIC and SOFA scores at different time points for sepsis prognosis was evaluated. RESULTS: In the non-survivor group, SIC and SOFA scores gradually increased during the first 4 days (P < 0.05). In the survivor group, the SIC and SOFA scores on day 2 were significantly higher than those on day 1 (P < 0.05); however, they decreased on day 4, dropping below the levels observed on day 1 (P < 0.05). The non-survivors showed higher SIC scores on days 2 (P < 0.05) and 4 (P < 0.001) than the survivors, whereas no significant differences were found between the two groups on day 1 (P > 0.05). The performance of SIC scores on day 4 for predicting mortality was more accurate than that on day 2, with areas under the curve of 0.749 (95% confidence interval [CI]: 0.674-0.823), and 0.601 (95% CI: 0.524-0.679), respectively. The SIC scores demonstrated comparable predictive accuracy for 28-day mortality to the SOFA scores on days 2 and 4. Cox proportional hazards models indicated that SIC on day 4 (hazard ratio [HR] = 3.736; 95% CI: 2.025-6.891) was an independent risk factor for 28-day mortality. CONCLUSIONS: The time course of SIC and SOFA scores differed between surviving and non-surviving patients with sepsis, and persistent high SIC and SOFA scores can predict 28-day mortality.

Acute muscle wasting is associated with poor prognosis in older adults with severe community-acquired pneumonia.

PURPOSE: To investigate the impact of acute muscle wasting on 90-day mortality in older patients with severe pneumonia using ultrasound and chest computed tomography (CT). METHODS: Quadriceps muscle layer thickness was measured via ultrasound on days 1, 7, and 14, and cross-sectional area of the erector spinae muscle was assessed using chest CT on days 1 and 14 in patients aged ≥ 65 years old. The primary outcome was all-cause 90-day mortality. Receiver operating characteristic curves were conducted for muscle loss to predict 90-day mortality. Cox proportional hazard models and Kaplan-Meier survival curves were employed to evaluate the association between muscle loss and 90-day mortality. RESULTS: Sixty-two patients were enrolled with median age of 80.2 years, 29 (46.8%) were men and 28 (45.2%) patients died. Muscle mass measured using ultrasound and CT decreased significantly from baseline to day 14 in the non-survivor group. Muscle loss assessed by ultrasound (with minimum and maximum pressure) and CT independently predicted all-cause 90-day mortality (adjusted hazard ratios = 1.497, 1.400 and 1.082; P < 0.001, P = 0.002, and P = 0.004; respectively), and cutoff values of muscle loss were 0.34 cm, 0.11 cm and 4.92 cm2, correspondingly. A higher muscle loss had an increased risk of 90-day mortality. CONCLUSIONS: Acute muscle wasting assessed by ultrasound and chest CT persisted for 14 days and was an independent predictor of adverse outcomes in older patients with severe pneumonia. A greater decline in muscle mass was associated with a higher 90-day mortality risk.

Predictive nomogram for in-hospital mortality among older patients with intra-abdominal sepsis incorporating skeletal muscle mass.

BACKGROUND: Studies on prognostic factors for older patients with intra-abdominal sepsis are scarce, and the association between skeletal muscle mass and prognosis among such patients remains unclear. AIMS: To develop a nomogram to predict in-hospital mortality among older patients with intra-abdominal sepsis. METHODS: Older patients with intra-abdominal sepsis were prospectively recruited. Their demographics, clinical features, laboratory results, abdominal computed tomography-derived muscle mass, and in-hospital mortality were recorded. The predictors of mortality were selected via least absolute shrinkage and selection operator and multivariable logistic regression analyses, and a nomogram was developed. The nomogram was assessed and compared with Sequential Organ Failure Assessment score, Acute Physiology and Chronic Health Evaluation II score, and Simplified Acute Physiology Score II. RESULTS: In total, 464 patients were included, of whom 104 (22.4%) died. Six independent risk factors (skeletal muscle index, cognitive impairment, frailty, heart rate, red blood cell distribution width, and blood urea nitrogen) were incorporated into the nomogram. The Hosmer-Lemeshow goodness-of-fit test and calibration plot revealed a good consistency between the predicted and observed probabilities. The area under the receiver operating characteristic curve was 0.875 (95% confidence interval = 0.838-0.912), which was significantly higher than those of commonly used scoring systems. The decision curve analysis indicated the nomogram had good predictive performance. DISCUSSION: Our nomogram, which is predictive of in-hospital mortality among older patients with intra-abdominal sepsis, incorporates muscle mass, a factor that warrants consideration by clinicians. The model has a high prognostic ability and might be applied in clinical practice after external validation.

Erector spinae muscle-based nomogram for predicting in-hospital mortality among older patients with severe community-acquired pneumonia.

BACKGROUND: No multivariable model incorporating erector spinae muscle (ESM) has been developed to predict clinical outcomes in older patients with severe community-acquired pneumonia (SCAP). This study aimed to construct a nomogram based on ESM to predict in-hospital mortality in patients with SCAP. METHODS: Patients aged ≥ 65 years with SCAP were enrolled in this prospective observational study. Least absolute selection and shrinkage operator and multivariable logistic regression analyses were used to identify risk factors for in-hospital mortality. A nomogram prediction model was constructed. The predictive performance was evaluated using the concordance index (C-index), calibration curve, net reclassification improvement (NRI), integrated discrimination improvement (IDI), and decision curve analysis. RESULTS: A total of 490 patients were included, and the in-hospital mortality rate was 36.1%. The nomogram included the following independent risk factors: mean arterial pressure, peripheral capillary oxygen saturation, Glasgow Coma Scale score (GCS), lactate, lactate dehydrogenase, blood urea nitrogen levels, and ESM cross-sectional area. Incorporating ESM into the base model with other risk factors significantly improved the C-index from 0.803 (95% confidence interval [CI], 0.761-0.845) to 0.836 (95% CI, 0.798-0.873), and these improvements were confirmed by category-free NRI and IDI. The ESM-based nomogram demonstrated a high level of discrimination, good calibration, and overall net benefits for predicting in-hospital mortality compared with the combination of confusion, urea, respiratory rate, blood pressure, and age ≥ 65 years (CURB-65), Pneumonia Severity Index (PSI), Acute Physiology and Chronic Health Evaluation II (APACHEII), and Sequential Organ Failure Assessment (SOFA). CONCLUSIONS: The proposed ESM-based nomogram for predicting in-hospital mortality among older patients with SCAP may help physicians to promptly identify patients prone to adverse outcomes. TRIAL REGISTRATION: This study was registered at www.chictr.org.cn (registration number Chi CTR-2300070377).

Prediction of Effector Proteins from Trichoderma longibrachiatum Through Transcriptome Sequencing.

Trichoderma longibrachiatum SMF2 is an important biocontrol strain isolated by our group that can promote plant growth and induce plant disease resistance. To further study its biocontrol mechanism, the effector proteins secreted by T. longibrachiatum SMF2 were analyzed through bioinformatics and transcriptome sequencing. Overall, 478 secretory proteins produced by T. longibrachiatum were identified, of which 272 were upregulated after treatment with plants. Functional annotation showed that 36 secretory proteins were homologous with different groups of effectors from pathogenic microorganisms. Moreover, the quantitative PCR results of six putative effector proteins were consistent with those of transcriptome sequencing. Taken together, these findings indicate that the secretory proteins secreted by T. longibrachiatum SMF2 may act as effectors to facilitate its own growth and colonization or to induce plant immunity response.

Circulating mid-regional proadrenomedullin is a predictor of mortality in patients with COVID-19: a systematic review and meta-analysis.

BACKGROUND: Although there is increasing understanding of the changes in the laboratory parameters of Coronavirus disease 2019 (COVID-19), the correlation between circulating Mid-regional Proadrenomedullin (MR-proADM) and mortality of patients with COVID-19 is not fully understood. In this study, we conducted a systematic review and meta-analysis to evaluate the prognostic value of MR-proADM in patients with COVID-19. METHODS: The PubMed, Embase, Web of Science, Cochrane Library, Wanfang, SinoMed and Chinese National Knowledge Infrastructure (CNKI) databases were searched from 1 January 2020 to 20 March 2022 for relevant literature. The Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) was used to assess quality bias, STATA was employed to pool the effect size by a random effects model, and potential publication bias and sensitivity analyses were performed. RESULTS: 14 studies comprising 1822 patients with COVID-19 met the inclusion criteria, there were 1145 (62.8%) males and 677 (31.2%) females, and the mean age was 63.8 ± 16.1 years. The concentration of MR-proADM was compared between the survivors and non-survivors in 9 studies and the difference was significant (P < 0.01), I2 = 46%. The combined sensitivity was 0.86 [0.73-0.92], and the combined specificity was 0.78 [0.68-0.86]. We drew the summary receiver operating characteristic (SROC) curve and calculated the area under curve (AUC) = 0.90 [0.87-0.92]. An increase of 1 nmol/L of MR-proADM was independently associated with a more than threefold increase in mortality (odds ratio (OR) 3.03, 95% confidence interval (CI) 2.26-4.06, I2 = 0.0%, P = 0.633). The predictive value of MR-proADM for mortality was better than many other biomarkers. CONCLUSION: MR-proADM had a very good predictive value for the poor prognosis of COVID-19 patients. Increased levels of MR-proADM were independently associated with mortality in COVID-19 patients and may allow a better risk stratification.

CgPG21 is involved in the degradation of the cell wall during the secretory cavity formation in Citrus grandis 'Tomentosa' fruits.

MAIN CONCLUSION: CgPG21 is mainly located in the cell wall, participates in the intercellular layer degradation of the cell wall during the formation of secretory cavity in the intercellular space-forming and lumen-expanding stages. The secretory cavity is a common structure in Citrus plants and is the main site for synthesis and accumulation of medicinal ingredients. The secretory cavity is formed in lysogenesis, when epithelial cells enter a process of programmed cell death. Pectinases are known to be involved in degradation of the cell wall during the cytolysis of secretory cavity cells, but the changes in cell structure, the dynamic characteristics of cell wall polysaccharides and the related genes regulating cell wall degradation are unclear. In this study, electron microscopy and cell wall polysaccharide-labeling techniques were used to study the main characteristics of cell wall degradation of the secreting cavity of Citrus grandis 'Tomentosa' fruits. At the same time, the full CDS length of the pectinase gene CgPG21 was cloned, encoding a protein composed of 480 amino acids. CgPG21 is mainly located in the cell wall, participates in the degradation of the intercellular layer of the cell wall during the development of the secretory cavity, and plays an important role in the formation of the secretory cavity in the intercellular space-forming and lumen-expanding stages. With the development of secretory cavity, the cell wall polysaccharides of epithelial cells gradually degrade. CgPG21 is mainly involved in the intercellular layer degradation.

Computed tomography-based body composition is associated with adverse clinical outcomes among older patients with sepsis in the emergency department.

PURPOSE: To investigate the association between body composition and adverse clinical outcomes in older patients with sepsis in the emergency department. METHODS: Body composition, including the skeletal muscle area, skeletal muscle index (SMI), mean skeletal muscle density (SMD), and intramuscular fat area, was measured at the level of the third lumbar vertebra (L3) on abdominal computed tomography scans. Clinical outcomes included 90-day mortality, 90-day readmission, and discharge to long-term care. According to sex-specific cut-off values of L3 SMI and SMD, patients were divided into low SMI, low SMD, both low SMI and low SMD, and neither low SMI nor low SMD groups. RESULTS: In total, 443 patients were included, 162 (36.6%) of whom died. Lower SMI and SMD, as continuous variables, were independent risk factors for 90-day mortality (adjusted hazard ratio [HR] = 0.947 and 0.963, respectively, both p < 0.001). Cut-off values of L3 SMI and L3 SMD were 32.24 cm2/m2 and 30.01 HU for men and 28.28 cm2/m2 and 28.20 HU for women, respectively. The both low SMI and low SMD group had an increased risk of 90-day mortality (adjusted HR=3.059, p < 0.001), 90-day readmission (adjusted odds ratio [OR]=2.859, p = 0.006), and discharge to long-term care (adjusted OR = 2.814, p = 0.007). CONCLUSIONS: Lower muscle mass and muscle quality, as measured by skeletal muscle index and density, were independent risk factors for mortality among older patients with sepsis in the emergency department. Furthermore, patients with both low muscle mass and quality had an increased risk of mortality, readmission, and discharge to long-term care.

Development of a cGMP-compliant process to manufacture donor-derived, CD45RA-depleted memory CD19-CAR T cells.

Autologous chimeric antigen receptor (CAR) T cells targeting the CD19 antigen have demonstrated a high complete response rate in relapsed/refractory B-cell malignancies. However, autologous CAR T cell therapy is not an option for all patients. Here we optimized conditions for clinical-grade manufacturing of allogeneic CD19-CAR T cells using CD45RA-depleted donor memory T cells (Tm) for a planned clinical trial. Tm were activated using the MACS GMP T Cell TransAct reagent and transduced in the presence of LentiBOOST with a clinical-grade lentiviral vector that encodes a 2nd generation CD19-CAR with a 41BB.zeta endodomain. Transduced T cells were transferred to a G-Rex cell culture device for expansion and harvested on day 7 or 8 for cryopreservation. The resulting CD19-CAR(Mem) T cells expanded on average 34.2-fold, and mean CAR expression was 45.5%. The majority of T cells were CD4+ and had a central memory or effector memory phenotype, and retained viral specificity. CD19-CAR(Mem) T cells recognized and killed CD19-positive target cells in vitro and had potent antitumor activity in an ALL xenograft model. Thus we have successfully developed a current good manufacturing practice-compliant process to manufacture donor-derived CD19-CAR(Mem) T cells. Our manufacturing process could be readily adapted for CAR(Mem) T cells targeting other antigens.

Comparison of three frailty screening instruments for prediction of adverse outcomes among older adults in the emergency department.

AIM: To compare the predictive abilities of the FRAIL scale (FS), frailty screening questionnaire (FSQ) and clinical frailty scale (CFS) for adverse outcomes in older adults in the emergency department. METHODS: In total, 317 older adults aged ≥65 years attending emergency department was screened for frailty using the FS, FSQ and CFS. Outcome measures included all-cause 28-day mortality and intensive care unit readmission. Cox proportional hazards model was used for survival comparison. Logistic regression was used to analyze risk factors for readmissions. In addition, we calculated the C-statistic, net reclassification improvement and integrated discrimination improvement to evaluate the predictive value of three scales. RESULTS: The prevalence of frailty was 55.2% (FS), 47.0% (FSQ) and 69.4% (CFS). Cox regression and logistic regression analysis revealed that frailty screening by FS, FSQ and CFS was an independent risk factor for all-cause 28-day mortality and 30- and 90-day readmission after adjustment. Incorporation of FS, FSQ and CFS into a basic model with other risk factors significantly improved C-statistic. For all-cause 28-day mortality, the model including FS had the highest C-statistic from 0.786 (95% confidence interval: 0.706-0.865) to 0.854 (95% confidence interval: 0.802-0.907) and the improvements in risk prediction were also confirmed by category-free net reclassification improvement and integrated discrimination improvement, suggesting FS was significantly better than CFS and FSQ. The three tools had a low predictive ability for readmission (all C-statistics <0.7). CONCLUSIONS: All three frailty scales showed a predictive ability for 28-day mortality and readmission but FS may be the most valid tool in the emergency department. Geriatr Gerontol Int ••; ••: ••-•• Geriatr Gerontol Int 2022; 22: 851-856.

A Structural and Bioinformatics Investigation of a Fungal Squalene Synthase and Comparisons with Other Membrane Proteins.

There is interest in the development of drugs to treat fungal infections due to the increasing threat of drug resistance, and here, we report the first crystallographic structure of the catalytic domain of a fungal squalene synthase (SQS), Aspergillus flavus SQS (AfSQS), a potential drug target, together with a bioinformatics study of fungal, human, and protozoal SQSs. Our X-ray results show strong structural similarities between the catalytic domains in these proteins, but, remarkably, using bioinformatics, we find that there is also a large, highly polar helix in the fungal proteins that connects the catalytic and membrane-anchoring transmembrane domains. This polar helix is absent in squalene synthases from all other lifeforms. We show that the transmembrane domain in AfSQS and in other SQSs, stannin, and steryl sulfatase, have very similar properties (% polar residues, hydrophobicity, and hydrophobic moment) to those found in the "penultimate" C-terminal helical domain in squalene epoxidase, while the final C-terminal domain in squalene epoxidase is more polar and may be monotopic. We also propose structural models for full-length AfSQS based on the bioinformatics results as well as a deep learning program that indicate that the C-terminus region may also be membrane surface-associated. Taken together, our results are of general interest given the unique nature of the polar helical domain in fungi that may be involved in protein-protein interactions as well as being a future target for antifungals.

[Clinical value of nutritional risk scores in patients with sepsis associated acute renal injury].

OBJECTIVE: To investigate the clinical value of nutritional indexes including body mass index (BMI), albumin (ALB), nutrition risk screening 2002 (NRS 2002) and the nutrition risk in critically ill score (NUTRIC) in 28-day prognosis of patients with sepsis related acute kidney injury (AKI). METHODS: A prospective cohort study was conducted. Patients with sepsis treated in the emergency intensive care unit (EICU) of China Rehabilitation Research Center from December 1, 2018 to December 1, 2020 were observed for 7 days. Patients with sepsis related AKI were enrolled in this study. The gender, age, BMI, basic diseases, shock, number of affected organs, length of hospital stay, ALB, mechanical ventilation (MV) and vasoactive drug use, sequential organ failure score (SOFA), rapid sequential organ failure score (qSOFA) and acute physiology and chronic health evaluation II (APACHE II) were recorded. The NRS 2002 score and NUTRIC score were calculated. Cox regression model was used to analyze the risk factors of 28-day mortality in patients with sepsis related AKI. The receiver operator characteristic curves (ROC curves) were drawn and the areas under the ROC curves (AUC) were calculated, and the value of BMI, ALB, NRS 2002 score and NUTRIC score was analyzed to predict 28-day mortality in patients with sepsis related AKI. Kaplan Meier survival curves were used to analyze the effects of NRS 2002 score and NUTRIC score stratification on the 28 day prognosis of patients with sepsis related AKI. RESULTS: A total of 140 patients with sepsis related AKI were enrolled, including 73 survival patients and 67 died patients within 28 days. The 28-day mortality was 47.9% (67/140). BMI in the survival group was significantly higher than that in the death group [kg/m2: 22.0 (19.5, 25.6) vs. 20.7 (17.3, 23.9), P < 0.05], and NRS 2002 score and NUTRIC score were significantly lower than those in the death group [NRS 2002 score: 5 (4, 6) vs. 7 (6, 7), NUTRIC score: 6 (5, 7) vs. 7 (6, 9), both P < 0.05]. The ALB of the survival group was slightly higher than that of the death group, but the difference was not statistically significant. Cox regression analysis showed that NRS 2002 score and NUTRIC score were independent risk factors for 28-day death in patients with sepsis related AKI. ROC curve analysis showed that NUTRIC score had the strongest predictive ability for 28-day death [AUC = 0.785, 95% confidence interval (95%CI) was 0.708-0.850], followed by NRS 2002 score (AUC = 0.728, 95%CI was 0.647-0.800), but there was no significant difference between them. Compared with NRS 2002 score, the predictive ability of BMI and ALB was poor. Kaplan-Meier curve analysis showed that the prognosis of patients with NRS 2002 score≥5 was significantly worse than that of patients with NRS 2002 score < 5 (28-day cumulative survival rate: 42.1% vs. 75.6%, Log-Rank test: χ2 = 11.884, P = 0.001), and the prognosis of patients with NUTRIC score ≥ 6 was significantly worse than that of patients with NUTRIC score < 6 (28-day cumulative survival rate: 40.4% vs. 86.1%, Log-Rank test: χ2 = 19.026, P = 0.000). CONCLUSIONS: Patients with sepsis related AKI have high nutritional risk. Both NRS 2002 score and NUTRIC score have good predictive value for the prognosis of patients with sepsis related AKI, while BMI and ALB have low predictive value. Due to the complex calculation of NUTRIC score, NRS 2002 score may be more suitable for emergency department.

[Application-oriented structure and function study of proteins: a review].

Repurpose of key catalytic reaction and reconstruction of metabolic pathways all depend on in-depth understanding of the relevant proteins in protein engineering, green biomanufacturing and synthetic biology. The rapid development of synthetic biotechnology calls for proteins with excellent performance to fulfill the requirement for engineering enzymes and strains. The key is to prepare large quantities of target proteins with high purity, and study the structure-function relationship quickly and accurately. In the past 10 years, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences has built a protein structure-function study platform, and studied many proteins of industrial significance. Progress has been made on the terpene synthase related to natural plant products synthesis, PET plastic degradation related enzyme, and enzymes involved in biomass conversion and utilization. The structure-function study of these proteins provided theoretical basis for further engineering. Development of protein structure-function research technologies will facilitate the research of synthetic biology and promote biomanufacturing.

Biodegradation of diphenyl ether herbicide lactofen by Bacillus sp. YS-1 and characterization of two initial degrading esterases.

The extensive use of the diphenyl ether herbicide lactofen in recent years has caused serious environmental problems. Therefore, detoxification and elimination of lactofen from the environment are urgently required. In this study, the lactofen-degrading strain Bacillus sp. YS-1 was isolated, which achieved a 97.6% degradation rate of 50 mg/L lactofen within 15 h. The ester bond of lactofen was hydrolyzed, which generated acifluorfen, and then, the nitro group was reduced to the amino group, which generated aminoacifluorfen. Finally, the amino group was acetylated, which formed acetylated aminoacifluorfen, a novel end product in the degradation of lactofen. The toxicity of acetylated aminoacifluorfen to the root and seedling growth of cucumber and sorghum was significantly decreased compared with that of lactofen. The two esterase genes rhoE and rapE, encoding two esterases responsible for lactofen hydrolysis to acifluorfen, were cloned and expressed. The amino acid sequences encoded by rhoE and rapE were 27.78% and 88.21% identical with known esterases, respectively. The optimum temperatures for RhoE and RapE degradation of lactofen were 35 °C and 25 °C, respectively, and both esterases displayed maximal activity at pH 8.0. Both RhoE and RapE prioritized the degradation of (S)-(+)-lactofen, (S)-(-)-quizalofop-ethyl, and (S)-(-)-diclofop-methyl. This study provided the resources of bacterial strain and hydrolyzing enzyme for the removal of lactofen from the environment and the bioremediation of herbicide-contaminated soil.