Tanshinone IIA protects human coronary artery endothelial cells from ferroptosis by activating the NRF2 pathway.

The pathogenesis of atherosclerosis is closely related to endothelial cell injury caused by lipid peroxidation-induced ferroptosis. Tanshinone IIA (TSA) protects endothelial tissues from damage. In this study, we investigated whether TSA exerts its protective effect on endothelial cells by inhibiting ferroptosis. Ferroptosis was induced in human coronary artery endothelial cells (HCAECs), and cells were treated with TSA. Morphological examination indicated that TSA exerted a significant protective effect on the HCAECs. This was further confirmed by LDH release and cell death detection assays. Flow cytometry revealed that TSA significantly reduced the excessive accumulation of total cellular ROS and lipid ROS caused by ferroptosis inducers. TSA also restored the reduction of glutathione (GSH), a potent and abundant reductant in cells. In addition, we found that TSA promoted the expression of NRF2, an essential player in response to oxidative stress, and its downstream genes. Immunofluorescent staining revealed that TSA promoted the nuclear translocation of NRF2. Increased nuclear translocation of NRF2 was validated by Western blot evaluation of cytoplasmic and nuclear protein extracts. Furthermore, NRF2 inhibition abolished the protective effects of TSA on HCAECs. These data demonstrate that TSA represses ferroptosis via activation of NRF2 in HCAECs.

Amentoflavone prevents ox-LDL-induced lipid accumulation by suppressing the PPARγ/CD36 signal pathway.

The formation of fat-laden foam cells plays an important role in the initiation and progression of atherosclerosis (AS). Amentoflavone (AF) is found in various traditional Chinese medicines, such as ginkgo biloba, which are used to treat cardiovascular diseases (CVDs). We aimed to explore the potential effects and mechanisms of AF on lipid accumulation, and its possible application in atherosclerotic cardiovascular disease (ASCVD). Cellular models of lipid accumulation were established by treatment of HUASMCs and THP-1 cells with oxidized low-density lipoprotein (ox-LDL). Cell viability, lipid accumulation, and ox-LDL uptake were assessed. Small interfering RNAs (siRNAs) and overexpression plasmids were used to reveal the hierarchical correlations of regulatory pathways. AF reduced the lipid accumulation and ox-LDL uptake induced by ox-LDL, and reduced the expression levels of cluster of differentiation 36 (CD36) and peroxisome proliferator-activated receptor gamma (PPARγ) proteins, while the expression level of ATP binding cassette subfamily A member 1 (ABCA1) increased. Knockdown of PPARγ or CD36 with siRNAs prevented ox-LDL-induced lipid accumulation. Overexpression of CD36 or PPARγ promoted the lipid accumulation induced by ox-LDL and eliminated the effect of AF on ox-LDL-induced lipid accumulation. Overall, AF prevents ox-LDL-induced lipid accumulation by suppressing the PPARγ/CD36 signaling pathway.

[Identification of Streptococcus mutans in carious patients' saliva samples by MALDI-TOF mass spectrometry].

OBJECTIVE: To identify Streptococcus mutans (S. mutans) in carious patients' saliva using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and to establish a faster and more accurate method to identify S. mutans. METHODS: In this study, a total of 90 carious patients from Department of Endodontics of Peking University School of Stomatology were recruited. All these patients' saliva was collected. After extracting the protein of the samples, they were identified by MALDI-TOF MS. The composite profile was analyzed using BioExplorer 1.0 software. The scores ≥ 25 were considered as S. mutans, whereas the scores <25 were as considered as non S. mutans. Finally, these results were compared with 16S rDNA sequencing to figure out the sensitivity and concordance rate, respectively. RESULTS: The sensitivity of MALDI-TOF MS was 96.0%, and the concordance rate compared with 16S rDNA sequencing was as high as 98.7%. CONCLUSION: MALDI-TOF MS is high throughput, rapid and easy to perform in comparison to other conventional methods. It has a high sensivity and concordance rate. Thus, MALDI-TOF MS can serve as an effective tool for identification of S. mutans.

Actin stimulates reduction of the MICAL-2 monooxygenase domain.

MICALs are large, multidomain flavin-dependent monooxygenases that use redox chemistry to cause actin to depolymerize. Little enzymology has been reported for MICALs, and none has been reported for MICAL-2, an enzyme vital for the proliferation of prostate cancer. The monooxygenase domains of MICALs resemble aromatic hydroxylases, but their substrate is the sulfur of a methionine of actin. In order to determine how closely MICAL-2 conforms to the aromatic hydroxylase paradigm, we studied its reaction with NAD(P)H. The enzyme has a strong preference for NADPH over NADH caused by a large difference in binding NADPH. A comparison of the reduction kinetics using protio-NADPH and [4R-(2)H]-NADPH showed that MICAL-2 is specific for the proR hydride of NADPH, as evidenced by a 4.8-fold kinetic isotope effect. The reductive half-reaction of the MICAL-2 hydroxylase domain is stimulated by f-actin. In the absence of actin, NADPH reduces the flavin relatively slowly; actin speeds that reaction significantly. The separate monooxygenase domain of MICAL-2 has the classic regulatory behavior of flavin-dependent aromatic hydroxylases (Class A monooxygenases): slow reduction of the flavin when the substrate to be oxygenated is absent. This prevents the wasteful consumption of reduced pyridine nucleotide and the production of harmful H2O2. Our results show that this strategy is used by MICAL-2. Thus, our data suggest that MICAL-2 could regulate catalysis through the monooxygenase domain alone; control by interactions with other domains of MICAL in the full-length enzyme may not be needed.

[Response Characteristics of Soil Organic Carbon Pool and Its Chemical Composition During Secondary Forest Succession in the Loess Plateau].

In order to explore the characteristics of the soil organic carbon(SOC)pool and its chemical composition during the succession of secondary forests in the Loess Plateau, samples of the primary stage (Populus davidiana forest), transition stage (Populus davidiana and Quercus wutaishansea mixed forest), and top stage (Quercus wutaishansea forest) of secondary forest succession in the Huanglong Mountain forest area of the Loess Plateau in Northern Shaanxi were selected as the research object. The variation characteristics of SOC content, storage, and its chemical composition at different soil depths (0-10, 10-20, 20-30, 30-50, and 50-100 cm) were analyzed. The results showed that:① the contents and storage of SOC increased significantly with the secondary forest succession process (P<0.05). The content of SOC decreased significantly with the increase in soil depth, and the storage of SOC increased from 64.8 Mg·hm-2 in the primary stage to 129.2 Mg·hm-2 in the top stage, with an increase of 99%. ② During the succession of secondary forests, in the surface (0-30 cm) soil organic carbon, the relative content of aliphatic carbon components that have a simple structure and can be decomposed more easily decreased, and the relative content of aromatic carbon components that have a complex structure and cannot be decomposed easily increased, indicating that the chemical composition of organic carbon stability of surface-layer soil increased significantly with the process of secondary forest succession. However, the stability of the chemical composition of SOC in the deep layer (30-100 cm) first increased and then decreased, that is, the transition stage>the top stage>the primary stage. ③In the process of secondary forest succession, the stability of SOC chemical composition in the primary stage and transition stage increased significantly with the increase in soil depth. The top stage tended to be stable, and the deep soil carbon stability decreased slightly. ④ Pearson correlation analysis showed that during the secondary forest succession process, SOC storage and chemical composition stability were significantly negatively correlated with soil total phosphorus content. In general, the content and storage of SOC in the 0-100 cm soil increased significantly during the secondary forest succession, playing the role of a "carbon sink." The stability of the chemical composition of SOC in the surface layer (0-30 cm) increased significantly, but in the deep layer (30-100 cm), it increased first and then decreased.

Dietary Guanidine Acetic Acid Addition Improved Carcass Quality with Less Back-Fat Thickness and Remarkably Increased Meat Protein Deposition in Rapid-Growing Lambs Fed Different Forage Types.

The aim of this study was to investigate whether guanidine acetic acid (GAA) yields a response in rapid-growing lambs depending on forage type. In this study, seventy-two small-tailed Han lambs (initial body weights = 12 ± 1.6 kg) were used in a 120-d feeding experiment after a 7-d adaptation period. A 2 × 3 factorial experimental feeding design was applied to the lambs, which were fed a total mixed ration with two forage types (OH: oaten hay; OHWS: oaten hay plus wheat silage) and three forms of additional GAA (GAA: 0 g/kg; UGAA: Uncoated GAA, 1 g/kg; CGAA: Coated GAA, 1 g/kg). The OH diet had a greater dry matter intake, average daily gain, and hot carcass weight than the OHWS diet. The GAA supplementation increased the final body weight, hot carcass weight, dressing percentage, and ribeye area in the longissimus lumborum. Meanwhile, it decreased backfat thickness and serum triglycerides. Dietary GAA decreased the acidity of the meat and elevated the water-holding capacity in mutton. In addition, the crude protein content in mutton increased with GAA addition. Dietary GAA (UGAA or CGAA) might be an effective additive in lamb fed by different forage types, as it has potential to improve growth performance and meat quality.

Randomized Trial of First-Line Tyrosine Kinase Inhibitor With or Without Radiotherapy for Synchronous Oligometastatic EGFR-Mutated Non-Small Cell Lung Cancer.

BACKGROUND: Adding radiotherapy (RT) to systemic therapy improves progression-free survival (PFS) and overall survival (OS) in oligometastatic non-small cell lung cancer (NSCLC). Whether these findings translate to epidermal growth factor receptor (EGFR)-mutated NSCLC remains unknown. The SINDAS trial (NCT02893332) evaluated first-line tyrosine kinase inhibitor (TKI) therapy for EGFR-mutated synchronous oligometastatic NSCLC and randomized to upfront RT vs no RT; we now report the prespecified interim analysis at 68% accrual. METHODS: Inclusion criteria were biopsy-proven EGFR-mutated adenocarcinoma (per amplification refractory mutation system or next generation sequencing), with synchronous (newly diagnosed, treatment naïve) oligometastatic (≤5 metastases; ≤2 lesions in any one organ) NSCLC without brain metastases. All patients received a first-generation TKI (gefitinib, erlotinib, or icotinib), and randomization was between no RT vs RT (25-40 Gy in 5 fractions depending on tumor size and location) to all metastases and the primary tumor/involved regional lymphatics. The primary endpoint (intention to treat) was PFS. Secondary endpoints included OS and toxicities. All statistical tests were 2-sided. RESULTS: A total of 133 patients (n = 65 TKI only, n = 68 TKI with RT) were enrolled (2016-2019). The median follow-up was 23.6 months. The respective median PFS was 12.5 months vs 20.2 months (P < .001), and the median OS was 17.4 months vs 25.5 months (P < .001) for TKI only vs TKI with RT. Treatment yielded no grade 5 events and a 6% rate of symptomatic grade 3-4 pneumonitis in the TKI with RT arm. Based on the efficacy results of this prespecified interim analysis, the ethics committee recommended premature cessation of this trial. CONCLUSIONS: As compared with a first-line TKI alone, addition of upfront local therapy using RT statistically significantly improved PFS and OS for EGFR-mutated NSCLC.

Guanidine acetic acid exhibited greater growth performance in younger (13-30 kg) than in older (30-50 kg) lambs under high-concentrate feedlotting pattern.

Guanidine acetic acid (GAA) is increasingly considered as a nutritional growth promoter in monogastric animals. Whether or not such response would exist in rapid-growing lambs is unclear yet. The objective of this study was to investigate whether dietary supplementation with uncoated GAA (UGAA) and coated GAA (CGAA) could alter growth performance, nutrient digestion, serum metabolites, and antioxidant capacity in lambs. Seventy-two small-tailed Han lambs initially weighed 12 ± 1.6 kg were randomly allocated into six groups in a 2 × 3 factorial experimental design including two forage-type rations [Oaten hay (OH) vs. its combination with wheat silage (OHWS)] and three GAA treatment per ration: no GAA, 1 g UGAA, and 1 g CGAA per kg dry matter. The whole experiment was completed in two consecutive growing stages (stage 1, 13-30 kg; stage 2, 30-50 kg). Under high-concentrate feeding pattern (Stage 1, 25: 75; Stage 2, 20: 80), UGAA or CGAA supplementation in young lambs presented greater dry matter intake (DMI) in stage 1 and average daily gain (ADG) in the whole experimental period; lambs in OH group had higher ADG and DMI than that in OHWS group in stage 1 and whole experimental period, but this phenomenon was not observed in stage 2. Both UCGA and CGAA addition increased dietary DM, organic matter (OM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) digestion in both stages. In blood metabolism, UCGA and CGAA addition resulted in a greater total protein (TP) and insulin-like growth factor 1(IGF-1) levels, as well as antioxidant capacity; at the same time, UCGA and CGAA addition increased GAA metabolism-creatine kinase and decreased guanidinoacetate N-methyltransferase (GAMT) and L-Arginine glycine amidine transferase catalyzes (AGAT) activity. In a brief, the results obtained in the present study suggested that GAA (UGAA and CGAA; 1 g/kg DM) could be applied to improve growth performance in younger (13-30 kg) instead of older (30-50 kg) lambs in high-concentrate feedlotting practice.

Is Intentional Replantation Appropriate for Treatment of Extensive Endodontic-periodontal Lesions Related to Palatogingival Groove?

This report describes the cases of two patients with a maxillary lateral incisor with palatogingival groove and extensive endodontic-periodontal lesions. Although it is reported that acceptable periodontal status is of great importance in case selection in intentional replantation, it is suggested in this report that intentional replantation could be chosen instead of immediate extraction if extensive endodontic-periodontal lesions exist in a tooth with palatogingival groove. The gingival margin position and gingival papilla were well preserved and the bone defect was almost completely repaired. This was beneficial to the aesthetic prosthodontic treatment and implantation, although external root resorption was observed.

Deciphering the evolution characteristics of extracellular microbial products from autotrophic and mixotrophic anammox consortia in response to nitrogen loading variations.

Extracellular microbial products (EMP) in biological wastewater treatment systems vary with operational conditions and in turn indicate the metabolic status of functional bacteria. In this study, the response of EMP from autotrophic and mixotrophic anammox consortia (AAC and MAC) to the variation of total nitrogen loading rates (TNLR) were investigated as well as their correlations with the community evolution. The variation of TNLR showed a significantly negative correlation with the production of bound microbial products (BMP) but a significantly positive correlation with the production of soluble microbial products (SMP). The presence of organic matters with COD/TN ratio of 0.15 limited the abundance of anammox bacteria in MAC at the full-load phase and suppressed their proliferation at the restart phase. Due to the improved abundance of carbohydrate metabolism genes, MAC with lower abundance of anammox bacteria produced lower soluble polysaccharides than AMC at the full-load phase. Furthermore, four components (C1-4) were identified on the excitation-emission matrix fluorescence spectra of SMP using parallel factor analysis. C1 exhibited a relative higher proportion at the full-load phase, whereas C4 was generated only at the light-load phase or empty-load phase. At the restart phase, C2 and C3 appeared simultaneously and accounted for a high proportion. The information of four components also suggested the metabolic status of AC as revealed by the specific anammox activity, which therefore provided a novel complementary but direct approach for monitoring the operation status of anammox bioreactors.

Achieving completely anaerobic ammonium removal over nitrite (CAARON) in one single UASB reactor: Synchronous and asynchronous feeding regimes of organic carbon make a difference.

At least 11% of total nitrogen (TN) remains in the anammox effluent, making it difficult to meet increasingly stringent discharge standards. To overcome this bottleneck, an innovative process to achieve completely anaerobic ammonium removal over nitrite (CAARON) in one single up-flow anaerobic sludge blanket reactor was proposed in this study. The synchronous feeding of acetate at a C/N (nitrite) ratio of 0.6 significantly reduced the nitrogen removal capacity of anammox reactor by limiting the abundance and metabolism of anammox bacteria. In contrast, the asynchronous feeding of acetate optimized the partition of the reactor column into two specific compartments: the lower half favoring anammox and the upper half dominated by DEAMOX (DEnitrifying AMmonium Oxidation). A high TN removal efficiency of 96.2±0.4% and a low effluent TN concentration of 9.3±0.9mgL-1 were obtained under a high TN loading rate of 9.0kgNm-3d-1. The dominant functional microbes in the CAARON process were identified as Candidatus Kuenenia and Thauera, which were responsible for the anammox and denitratation reactions, respectively. Overall, the results in this study provide valuable insight into the coupling of anammox with denitratation, which is a cost-efficient approach for treating ammonium-rich wastewaters.

Evaluating the effects of Zn(II) on high-rate biogranule-based denitrification: Performance, microbial community and sludge characteristics.

High-rate denitrification is a popular and efficient process for treatment of nitrate-rich wastewater. Knowing the effect of heavy metals on denitrification is essential for industrial development. In the present study, the long-term impacts of Zn(II) on denitrifying granular sludge were investigated. The suppression threshold of Zn(II) on denitrifying bacteria was 10 mg L-1 for long-term exposure. The nitrogen removal rate was decreased by long-term addition of 10 mg L-1 Zn(II). Castellaniella and Klebsiella were the two dominant genera under Zn(II) stress. The relative abundance of Klebsiella sharply decreased to 4.64% after the addition of 10 mg L-1 Zn(II), whereas Castellaniella was susceptible to 2 mg L-1 Zn(II), revealing that Castellaniella mainly was devoted to denitrification under no or low Zn(II) stress conditions, whereas Klebsiella was effective under high Zn(II) stress.

The revolution of performance, sludge characteristics and microbial community of anammox biogranules under long-term NiO NPs exposure.

Given the increasing applications of NiO nanoparticles (NPs) in battery products, the potential effects of NiO NPs on anaerobic ammonium oxidation (anammox) systems were studied for the first time. The results showed that the anammox system performance obviously differed under the stresses of different NiO NPs concentrations. After the withdrawal of NiO NPs, the nitrogen removal performance of the anammox reactor returned to nearly that of the initial phase within 35 days. Compared with 0 mg L-1 NiO NPs, the specific anammox activity first increased and then decreased to the minimum value of 116.8 ±â€¯13.8 mg TN g-1 VSS d-1 at 60 mg L-1 NiO NPs. The variations in the heme c contents and extracellular polymeric substance amounts were similar to the variations in the specific anammox activity throughout the whole experiment. Additionally, the relative abundance of the dominant bacteria (Candidatus kuenenia) increased from 20.44% at 60 mg L-1 NiO NPs to 23.14% at the end of the last phase. Thus, the potential effects of NiO NPs on anammox systems should be a cause for great concern.

Effects of ZnO nanoparticles on high-rate denitrifying granular sludge and the role of phosphate in toxicity attenuation.

The increasing release of engineered nanoparticles (NPs) from consumer products has raised great concerns about their impacts on biological wastewater treatment. In this study, the widely-used ZnO NP was selected as a model NP to investigate its impact on high-rate denitrifying granular sludge in terms of sludge properties and community structure. A hormesis effect was observed during short-term exposure, in which the specific denitrification activity (SDA) was stimulated by 10% at 1 mg L-1 ZnO NPs, but inhibited by 23% at 5.0 mg L-1 ZnO NPs. When continuously exposed to 2.5 mg L-1 ZnO NPs, the nitrogen removal capacity of the denitrification reactor was nearly deprived within 15 days, and the relative abundance of the dominant denitrifying bacterium (Castellaniella) was decreased from 51.0 to 8.0%. Meanwhile, the dehydrogenase activity (DHA) and the content of extracellular polymeric substance (EPS) significantly decreased to 22.3 and 61.1%, respectively. Nevertheless, the presence of phosphate substantially weakened the adverse effects of ZnO NPs on the SDA, EPS, DHA and the relative abundance of functional genes even exposed to 6.25 mg L-1 ZnO NPs, which was associated with the fact that the level of Zn(II) released from ZnO NPs was significantly reduced in the presence of phosphate. Therefore, the toxicity of ZnO NPs may be mainly attributed to the release of toxic Zn(II) and could be attenuated in the presence of phosphate. Overall, this study provided further reference and meaningful insights into the impact of engineered NPs on biological wastewater treatment.

l-Carnitine and heart disease.

Cardiovascular disease (CVD) is a key cause of deaths worldwide, comprising 15-17% of healthcare expenditure in developed countries. Current records estimate an annual global average of 30 million cardiac dysfunction cases, with a predicted escalation by two-three folds for the next 20-30years. Although ß-blockers and angiotensin-converting-enzymes are commonly prescribed to control CVD risk, hepatotoxicity and hematological changes are frequent adverse events associated with these drugs. Search for alternatives identified endogenous cofactor l-carnitine, which is capable of promoting mitochondrial ß-oxidation towards a balanced cardiac energy metabolism. l-Carnitine facilitates transport of long-chain fatty acids into the mitochondrial matrix, triggering cardioprotective effects through reduced oxidative stress, inflammation and necrosis of cardiac myocytes. Additionally, l-carnitine regulates calcium influx, endothelial integrity, intracellular enzyme release and membrane phospholipid content for sustained cellular homeostasis. Carnitine depletion, characterized by reduced expression of "organic cation transporter-2" gene, is a metabolic and autosomal recessive disorder that also frequently associates with CVD. Hence, exogenous carnitine administration through dietary and intravenous routes serves as a suitable protective strategy against ventricular dysfunction, ischemia-reperfusion injury, cardiac arrhythmia and toxic myocardial injury that prominently mark CVD. Additionally, carnitine reduces hypertension, hyperlipidemia, diabetic ketoacidosis, hyperglycemia, insulin-dependent diabetes mellitus, insulin resistance, obesity, etc. that enhance cardiovascular pathology. These favorable effects of l-carnitine have been evident in infants, juvenile, young, adult and aged patients of sudden and chronic heart failure as well. This review describes the mechanism of action, metabolism and pharmacokinetics of l-carnitine. It specifically emphasizes upon the beneficial role of l-carnitine in cardiomyopathy.

Radiographic healing after a root canal treatment performed in single-rooted teeth with and without ultrasonic activation of the irrigant: a randomized controlled trial.

INTRODUCTION: The aim of this study was to compare the outcome of a root canal treatment with and without additional ultrasonic activation of the irrigant. METHODS: Single-rooted teeth with radiographic evidence of periapical bone loss were randomly assigned to 2 treatment groups. In both groups syringe irrigation was performed, and in one group the irrigant was also activated by ultrasound. Ten to 19 months after treatment, the teeth were examined by using periapical radiography (PA) and cone-beam computed tomography (CBCT). Area and volume of the periapical lesions were measured, and the outcome was presented in 4 categories: absence, reduction or enlargement of the radiolucency, or uncertain. Lesions were classified as reduced or enlarged when the change in size of the radiolucency was 20% or more. RESULTS: The recall rate was 82%, and 84 teeth were analyzed. CBCT detected significantly more post-treatment lesions than PA (P = .038), but the percentages of absence and reduction of the radiolucency together revealed by CBCT and PA were similar (P = .383). The CBCT results showed that absence of the radiolucency was observed in 16 of 84 teeth (19%) and reduction of the radiolucency in 61 of 84 teeth (72.6%), but there was no significant difference between the results of the 2 groups (P = .470). Absence and reduction of the radiolucency together were observed in the ultrasonic group in 39 of 41 teeth (95.1%) and in the syringe group in 38 of 43 teeth (88.4%). CONCLUSIONS: Root canal treatments with and without additional ultrasonic activation of the irrigant contributed equally to periapical healing.

Heart rate dependence of aortic pulse wave velocity at different arterial pressures in rats.

Arterial stiffness, as measured by aortic pulse wave velocity (PWV), is an independent marker of cardiovascular disease and events in both healthy and diseased populations. Although some cardiovascular risk factors, such as age and blood pressure, show a strong association with PWV, the association between heart rate (HR) and PWV is not firmly established. Furthermore, this association has not been investigated at different arterial blood pressures. To study effects of HR on aortic PWV at different mean arterial pressures (MAPs), adult (12 weeks; n=7), male, anesthetized Sprague-Dawley rats were randomly paced at HRs of between 300 and 450 bpm, at 50-bpm steps. At each pacing step, aortic PWV was measured across a physiological MAP range of 60 to 150 mmHg by infusing sodium nitroprusside and phenylephrine. When compared at the same MAP, increases in HR resulted in significant increases in PWV at all of the MAPs >80 mmHg (ANOVA, P<0.05), with the greatest significant change of 6.03±0.93% observed in the range 110 to 130 mmHg. The positive significant association between HR and PWV remained when PWV was adjusted for MAP (ANOVA, P<0.001). These results indicate that HR dependency of PWV is different at higher pressures than at lower pressures and that HR may be a confounding factor that should be taken into consideration when performing analysis based on PWV measurements.