Structure-based discovery of CFTR potentiators and inhibitors.

The cystic fibrosis transmembrane conductance regulator (CFTR) is a crucial ion channel whose loss of function leads to cystic fibrosis, whereas its hyperactivation leads to secretory diarrhea. Small molecules that improve CFTR folding (correctors) or function (potentiators) are clinically available. However, the only potentiator, ivacaftor, has suboptimal pharmacokinetics and inhibitors have yet to be clinically developed. Here, we combine molecular docking, electrophysiology, cryo-EM, and medicinal chemistry to identify CFTR modulators. We docked ∼155 million molecules into the potentiator site on CFTR, synthesized 53 test ligands, and used structure-based optimization to identify candidate modulators. This approach uncovered mid-nanomolar potentiators, as well as inhibitors, that bind to the same allosteric site. These molecules represent potential leads for the development of more effective drugs for cystic fibrosis and secretory diarrhea, demonstrating the feasibility of large-scale docking for ion channel drug discovery.

Conformational Changes of CFTR upon Phosphorylation and ATP Binding.

The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel evolved from an ATP-binding cassette transporter. CFTR channel gating is strictly coupled to phosphorylation and ATP hydrolysis. Previously, we reported essentially identical structures of zebrafish and human CFTR in the dephosphorylated, ATP-free form. Here, we present the structure of zebrafish CFTR in the phosphorylated, ATP-bound conformation, determined by cryoelectron microscopy to 3.4 Å resolution. Comparison of the two conformations shows major structural rearrangements leading to channel opening. The phosphorylated regulatory domain is disengaged from its inhibitory position; the nucleotide-binding domains (NBDs) form a "head-to-tail" dimer upon binding ATP; and the cytoplasmic pathway, found closed off in other ATP-binding cassette transporters, is cracked open, consistent with CFTR's unique channel function. Unexpectedly, the extracellular mouth of the ion pore remains closed, indicating that local movements of the transmembrane helices can control ion access to the pore even in the NBD-dimerized conformation.

Molecular Structure of the Human CFTR Ion Channel.

The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that uniquely functions as an ion channel. Here, we present a 3.9 Å structure of dephosphorylated human CFTR without nucleotides, determined by electron cryomicroscopy (cryo-EM). Close resemblance of this human CFTR structure to zebrafish CFTR under identical conditions reinforces its relevance for understanding CFTR function. The human CFTR structure reveals a previously unresolved helix belonging to the R domain docked inside the intracellular vestibule, precluding channel opening. By analyzing the sigmoid time course of CFTR current activation, we propose that PKA phosphorylation of the R domain is enabled by its infrequent spontaneous disengagement, which also explains residual ATPase and gating activity of dephosphorylated CFTR. From comparison with MRP1, a feature distinguishing CFTR from all other ABC transporters is the helix-loop transition in transmembrane helix 8, which likely forms the structural basis for CFTR's channel function.

The transcription factor EBF1 non-cell-autonomously regulates cardiac growth and differentiation.

Reciprocal interactions between non-myocytes and cardiomyocytes regulate cardiac growth and differentiation. Here, we report that the transcription factor Ebf1 is highly expressed in non-myocytes and potently regulates heart development. Ebf1-deficient hearts display myocardial hypercellularity and reduced cardiomyocyte size, ventricular conduction system hypoplasia, and conduction system disease. Growth abnormalities in Ebf1 knockout hearts are observed as early as embryonic day 13.5. Transcriptional profiling of Ebf1-deficient embryonic cardiac non-myocytes demonstrates dysregulation of Polycomb repressive complex 2 targets, and ATAC-Seq reveals altered chromatin accessibility near many of these same genes. Gene set enrichment analysis of differentially expressed genes in cardiomyocytes isolated from E13.5 hearts of wild-type and mutant mice reveals significant enrichment of MYC targets and, consistent with this finding, we observe increased abundance of MYC in mutant hearts. EBF1-deficient non-myocytes, but not wild-type non-myocytes, are sufficient to induce excessive accumulation of MYC in co-cultured wild-type cardiomyocytes. Finally, we demonstrate that BMP signaling induces Ebf1 expression in embryonic heart cultures and controls a gene program enriched in EBF1 targets. These data reveal a previously unreported non-cell-autonomous pathway controlling cardiac growth and differentiation.

Genome-wide identification of the Q-type C2H2 zinc finger protein gene family and expression analysis under abiotic stress in lotus (Nelumbo nucifera G.).

BACKGROUND: Lotus (Nelumbo nucifera G.) is an important aquatic plant with high ornamental, economic, cultural and ecological values, but abiotic stresses seriously affect its growth and distribution. Q-type C2H2 zinc finger proteins (ZFPs) play an important role in plant growth development and environmental stress responses. Although the Q-type C2H2 gene family has been identified in some plants, limited reports has been carried out it in lotus. RESULTS: In this study, we identified 45 Q-type NnZFP members in lotus. Based on the phylogenetic tree, these Q-type NnZFP gene family members were divided into 4 groups, including C1-1i, C1-2i, C1-3i and C1-4i. Promoter cis-acting elements analysis indicated that most Q-type NnZFP gene family members in lotus were associated with response to abiotic stresses. Through collinearity analyses, no tandem duplication gene pairs and 14 segmental duplication gene pairs were identified, which showed that duplication events might play a key role in the expansion of the Q-type NnZFP gene family. The synteny results suggested that 54 and 28 Q-type NnZFP genes were orthologous to Arabidopsis and rice, respectively. The expression patterns of these Q-type NnZFP genes revealed that 30 Q-type NnZFP genes were expressed in at least one lotus tissue. Nn5g30550 showed relatively higher expression levels in all tested tissues. 12 genes were randomly selected with at least one gene from each phylogenetic clade, and the expression of these selected genes were confirmed by qRT-PCR (quantitative real-time polymerase chain reaction). The results indicated that Q-type NnZFP genes were extensively involved in cadmium, drought, salt and cold stresses responses. Among them, 11 genes responded to at least three different stress treatments, especially Nn2g12894, which induced by all four treatments. CONCLUSIONS: These results could increase our understanding of the characterization of the Q-type NnZFP gene family and provide relevant information for further functional analysis of Q-type NnZFP genes in plant development, and abiotic stress tolerance in lotus.

Involvement of CD44 and MAPK14-mediated ferroptosis in hemorrhagic shock.

To elucidate the induction of ferroptotic pathways and the transcriptional modulation of pivotal genes in the context of hemorrhagic shock. The R software was used to analyze the GSE64711 dataset, isolating genes relevant to ferroptosis. Enrichment analyses and protein interaction networks were assembled. Using WGCNA hub genes were identified and intersected with ferroptosis-related genes, highlighting hub genes CD44 and MAPK14. In a rat hemorrhagic shock model, cardiac ROS, Fe2+, MDA, and GSH levels were assessed. Key ferroptotic proteins (SLC7A11/GPX4) in myocardial tissues were examined via western blot. Hub genes, CD44 and MAPK14, expressions were confirmed through immunohistochemistry. Analyzing the GSE64711 dataset revealed 337 differentially expressed genes, including 12 linked to ferroptosis. Enrichment analysis highlighted pathways closely related to ferroptosis. Using Genemania, we found these genes mainly affect ROS metabolism and oxidative stress response. WGCNA identified CD44 and MAPK14 as hub genes. Rat myocardial tissue validation showed significant cardiac damage and elevated ROS and MDA levels, and decreased GSH levels in the hemorrhagic shock model. The ferroptotic pathway SLC7A11/GPX4 was activated, and immunohistochemistry showed a significant increase in the expression levels of CD44 and MAPK14 in the hemorrhagic shock rat model. We demonstrated the presence of tissue ferroptosis in hemorrhagic shock by combining bioinformatics analysis with in vivo experimentation. Specifically, we observed the activation of the SLC7A11/GPX4 ferroptotic pathway. Further, CD44 and MAPK14 were identified as hub genes in hemorrhagic shock.

Neural cell adhesion molecule is required for ventricular conduction system development.

The most distal portion of the ventricular conduction system (VCS) contains cardiac Purkinje cells (PCs), which are essential for synchronous activation of the ventricular myocardium. Contactin-2 (CNTN2), a member of the immunoglobulin superfamily of cell adhesion molecules (IgSF-CAMs), was previously identified as a marker of the VCS. Through differential transcriptional profiling, we discovered two additional highly enriched IgSF-CAMs in the VCS: NCAM-1 and ALCAM. Immunofluorescence staining showed dynamic expression patterns for each IgSF-CAM during embryonic and early postnatal stages, but ultimately all three proteins became highly enriched in mature PCs. Mice deficient in NCAM-1, but not CNTN2 or ALCAM, exhibited defects in PC gene expression and VCS patterning, as well as cardiac conduction disease. Moreover, using ST8sia2 and ST8sia4 knockout mice, we show that inhibition of post-translational modification of NCAM-1 by polysialic acid leads to disrupted trafficking of sarcolemmal intercalated disc proteins to junctional membranes and abnormal expansion of the extracellular space between apposing PCs. Taken together, our data provide insights into the complex developmental biology of the ventricular conduction system.

Prognostic value and immunological role of PD-L1 gene in pan-cancer.

OBJECTIVE: PD-L1, a target of immune checkpoint blockade, has been proven to take the role of an oncogene in most human tumors. However, the role of PD-L1 in human pan-cancers has not yet been fully investigated. MATERIALS AND METHODS: Pan-cancer analysis was conducted to analyze expression, genetic alterations, prognosis analysis, and immunological characteristics of PD-L1. Estimating the correlation between PD-L1 expression and survival involved using pooled odds ratios and hazard ratios with 95% CI. The Kaplan-Meier (K-M) technique, COX analysis, and receiver operating characteristic (ROC) curves were applied to the survival analysis. Additionally, we investigated the relationships between PD-L1 and microsatellite instability (MSI), tumor mutational burden (TMB), DNA methyltransferases (DNMTs), the associated genes of mismatch repair (MMR), and immune checkpoint biomarkers using Spearman's correlation analysis. Also, immunohistochemical analysis and qRT-PCR were employed in evaluating PD-L1's protein and mRNA expression in pan-caner. RESULTS: PD-L1 showed abnormal mRNA and protein expression in a variety of cancers and predicted prognosis in cancer patients. Furthermore, across a variety of cancer types, the aberrant PD-L1 expression was connected to the MSI, MMR, TMB, drug sensitivity, and tumor immune microenvironment (TIME). Moreover, PD-L1 was significantly correlated with infiltrating levels of immune cells (T cell CD8 + , neutrophil, and so on). CONCLUSION: Our study provides a better theoretical basis and guidance for the clinical treatment of PD-L1.

Quantitative imaging for predicting hematoma expansion in intracerebral hemorrhage: A multimodel comparison.

BACKGROUND: Several studies report that radiomics provides additional information for predicting hematoma expansion in intracerebral hemorrhage (ICH). However, the comparison of diagnostic performance of radiomics for predicting revised hematoma expansion (RHE) remains unclear. METHODS: The cohort comprised 312 consecutive patients with ICH. A total of 1106 radiomics features from seven categories were extracted using Python software. Support vector machines achieved the best performance in both the training and validation datasets. Clinical factors models were constructed to predict RHE. Receiver operating characteristic curve analysis was used to assess the abilities of non-contrast computed tomography (NCCT) signs, radiomics features, and combined models to predict RHE. RESULTS: We finally selected the top 21 features for predicting RHE. After univariate analysis, 4 clinical factors and 5 NCCT signs were selected for inclusion in the prediction models. In the training and validation dataset, radiomics features had a higher predictive value for RHE (AUC = 0.83) than a single NCCT sign and expansion-prone hematoma. The combined prediction model including radiomics features, clinical factors, and NCCT signs achieved higher predictive performances for RHE (AUC = 0.88) than other combined models. CONCLUSIONS: NCCT radiomics features have a good degree of discrimination for predicting RHE in ICH patients. Combined prediction models that include quantitative imaging significantly improve the prediction of RHE, which may assist in the risk stratification of ICH patients for anti-expansion treatments.

Closed-Loop Recyclable Poly(ester-disulfide)s for Potential Alternatives to Engineering Plastic.

Facile fabrication, low material complexity and closed-loop recycling are essential for polymer plastics to alter their linear product economy towards a cradle-to-cradle one. Covalent adaptable networks (CANs) are one way to achieve that, which intrinsically exhibit decent mechanical properties like the thermosets but could also be easily recycled like the thermoplastics. In this work, we introduce rigid ester structural motifs into dynamic poly(disulfide)s to form a series of dual polymer networks. Owning to the coherence of soft/rigid segments and the reversible sacrificial crosslinking, they exhibit tailorable mechanical properties and good resistance towards different chemicals. Their closed-loop recycling is achieved via mild solvolysis, maintaining materials' mechanical integrities. It offers a solution as a sustainable replacement for engineering plastics which are massively under production but hard to be recycled.

Autofocusing of Fresnel zone aperture lensless imaging for QR code recognition.

Fresnel zone aperture (FZA) lensless imaging encodes the incident light into a hologram-like pattern, so that the scene image can be numerically focused at a long imaging range by the back propagation method. However, the target distance is uncertain. The inaccurate distance causes blurs and artifacts in the reconstructed images. This brings difficulties for the target recognition applications, such as quick response code scanning. We propose an autofocusing method for FZA lensless imaging. By incorporating the image sharpness metrics into the back propagation reconstruction process, the method can acquire the desired focusing distance and reconstruct noise-free high-contrast images. By combining the Tamura of the gradient metrics and nuclear norm of gradient, the relative error of estimated object distance is only 0.95% in the experiment. The proposed reconstruction method significantly improves the mean recognition rate of QR code from 4.06% to 90.00%. It paves the way for designing intelligent integrated sensors.

Moderate Ischemic Mitral Regurgitation with Ejection Fraction <40% Undergoing Concomitant Mitral Valve Repair during Revascularization: A Single-Center Observational Study.

Background: Numerous studies have examined the therapeutic effects of mitral valve repair during revascularization on moderate ischemic mitral regurgitation (IMR), as well as the incremental benefit of subvalvular repair alongside an annuloplasty ring. However, the impact of depressed left ventricular (LV) function on the surgical outcome of patients with moderate IMR has been rarely investigated. The aims of this single-center, retrospective, observational study were firstly to evaluate short- and medium-term outcomes in this patient group after undergoing mitral valve repair during revascularization, and secondly to assess the impact of depressed LV function on surgical outcomes. Methods: A total of 272 eligible patients who had moderate IMR and underwent concomitant mitral valve repair and revascularization from January 2010 to December 2017 were included in the study. These patients were categorized into different groups based on their ejection fraction (EF) levels: an EF < 40% group (n = 90) and an EF ≥ 40% group (n = 182). The median time course of follow-up was 42 months and the shortest follow-up time was 30 months. This study compared in-hospital outcomes (major postoperative morbidity and surgical mortality) as well as midterm outcomes (moderate or more mitral regurgitation, all-cause mortality, and reoperation) of the two groups before and after propensity score (PS) matching (1:1). Results: No significant difference was observed in surgical mortality between groups (8.9% vs. 3.3%, p = 0.076). More patients in the EF < 40% group developed low cardiac output (8.9% vs. 2.7%, p = 0.034) and prolonged ventilation (13.3% vs. 5.5%, p = 0.026) compared to the EF ≥ 40% group. Propensity score (PS) matching successfully established 82 patient pairs in a 1:1 ratio. No significance was discovered between the matched cohorts in terms of major postoperative morbidity and surgical mortality, except for prolonged ventilation. Conditional mixed-effects logistic regression analysis revealed that EF < 40% had an independent impact on prolonged ventilation (odds ratio (OR) = 2.814, 95% CI 1.321-6.151, p = 0.031), but was not an independent risk factor for surgical mortality (OR = 2.967, 95% CI 0.712-7.245, p = 0.138) or other major postoperative morbidity. Furthermore, the two groups showed similar cumulative survival before (log-rank p = 0.278) and after (stratified log-rank p = 0.832) PS matching. Cox regression analysis suggested that EF < 40% was not related to mortality compared with EF ≥ 40% (PS-adjusted hazard ratio (HR) = 1.151, 95% CI 0.763-1.952, p = 0.281). Conclusions: Patients with moderate IMR and EF < 40% shared similar midterm outcomes and surgical mortality to patients with moderate IMR and EF ≥ 40%, but received prolonged ventilation more often. Depressed LV function may be not associated with surgical or midterm mortality.

Portal vein thrombosis after cetuximab and 5-fluorouracil therapy in a patient with advanced colon cancer and decompensated cirrhosis: a case report and review of the literature.

BACKGROUND: Treatment options for advanced colon cancer are mainly combinations of chemotherapy and targeted drugs. However, poor physical health and medication intolerance limit the choice of anticancer drugs. Colon cancer with cirrhosis is a particular patient group that poses a challenge to clinical treatment. CASE PRESENTATION: This article presents a case of a patient in the decompensated stage of cirrhosis who was diagnosed with advanced colon cancer. The initial presentation was a nodule on his navel named the Sister Mary Joseph's nodule, which was later confirmed by biopsy and PET-CT as one of the metastases of colon cancer. The patient was treated with cetuximab and 5-fluorouracil at a below-guideline dose; however, portal vein thrombosis developed and led to death. This entire process, from diagnosis to death, occurred within a span of three months. CONCLUSION: Cancers with cirrhosis are a special group that deserves more attention. There is no unified treatment guideline for these patients, especially those with extrahepatic primary tumors. We should be more cautious when choosing treatment for such patients in the future. Both chemotherapy and targeted treatment may potentially induce portal vein thrombosis, which appears to have a higher incidence and worse prognosis than cancers without cirrhosis.

Wrist-Worn Accelerometry, Aging, and Gait Speed in the Baltimore Longitudinal Study of Aging.

Wrist-worn accelerometry metrics are not well defined in older adults. Accelerometry data from 720 participants (mean age 70 years, 55% women) were summarized into (a) total activity counts per day, (b) active minutes per day, (c) active bouts per day, and (d) activity fragmentation (the reciprocal of the mean active bout length). Linear regression and mixed-effects models were utilized to estimate associations between age and gait speed with wrist accelerometry. Activity counts per day, daily active minutes per day, and active bouts per day were negatively associated with age among all participants, while positive associations with activity fragmentation were only observed among those ≥65 years. More activity counts, more daily active minutes, and lower activity fragmentation were associated with faster gait speed. There were baseline age interactions with annual changes in total activity counts per day, active minutes per day, and activity fragmentation (Baseline age × Time, p < .01 for all). These results help define and characterize changes in wrist-based physical activity patterns among older adults.

[Exploration and example interpretation of real-world herbal prescription classification based on similarity matching algorithm].

In observational studies, herbal prescriptions are usually studied in the form of "similar prescriptions". At present, the classification of prescriptions is mainly based on clinical experience judgment, but there are some problems in manual judgment, such as lack of unified criteria, labor consumption, and difficulty in verification. In the construction of a database of integrated traditional Chinese and western medicine for the treatment of coronavirus disease 2019(COVID-19), our research group tried to classify real-world herbal prescriptions using a similarity matching algorithm. The main steps include 78 target prescriptions are determined in advance; four levels of importance labeling shall be carried out for the drugs of each target prescription; the combination, format conversion, and standardization of drug names of the prescriptions to be identified in the herbal medicine database; calculate the similarity between the prescriptions to be identified and each target prescription one by one; prescription discrimination is performed based on the preset criteria; remove the name of the prescriptions with "large prescriptions cover the small". Through the similarity matching algorithm, 87.49% of the real prescriptions in the herbal medicine database of this study can be identified, which preliminarily proves that this method can complete the classification of herbal prescriptions. However, this method does not consider the influence of herbal dosage on the results, and there is no recognized standard for the weight of drug importance and criteria, so there are some limitations, which need to be further explored and improved in future research.

Assessment of Physical Activity in Adults Using Wrist Accelerometers.

The health benefits of physical activity (PA) have been widely recognized, yet traditional measures of PA, including questionnaires and category-based assessments of volume and intensity, provide only broad estimates of daily activities. Accelerometers have advanced epidemiologic research on PA by providing objective and continuous measurement of PA in free-living conditions. Wrist-worn accelerometers have become especially popular because of low participant burden. However, the validity and reliability of wrist-worn devices for adults have yet to be summarized. Moreover, accelerometer data provide rich information on how PA is accumulated throughout the day, but only a small portion of these rich data have been used by researchers. Last, new methodological developments are emerging that aim to overcome some of the limitations of accelerometers. In this review, we provide an overview of accelerometry research, with a special focus on wrist-worn accelerometers. We describe briefly how accelerometers work; summarize the validity and reliability of wrist-worn accelerometers; discuss the benefits of accelerometers, including measuring light-intensity PA; and discuss pattern metrics of daily PA recently introduced in the literature. A summary of large-scale cohort studies and randomized trials that implemented wrist-worn accelerometry is provided. We conclude the review by discussing new developments and directions of research using accelerometers, with a focus on wrist-worn accelerometers.

Immediate Recognition and Surgical Treatment of Iatrogenic Acute Type A Aortic Dissection Is Associated with Low Hospital Mortality and High Intermediate-Term Survival.

Background: To investigate short- and intermediate-term outcomes of immediate (on table) recognition and surgical treatment of iatrogenic acute type A aortic dissection (ATAD) that occurred during the course of the cardiac surgical procedures. Methods: Of 23,143 adult patients undergoing cardiac surgical procedures at our institution from January 2016 to December 2020, 21 (0.09%) suffered from intraoperative iatrogenic ATAD and underwent immediate aortic repair. Their clinical characteristics, in-hospital outcomes and follow-up results were analyzed. Results: Among the 21 patients, 13 (61.9%) suffered from hypertension, and 14 (66.7%) had a dilated ascending aorta. In-hospital mortality was 9.5%, and new onset of permanent neurologic deficit was recorded in one patient. During a median follow-up of 36.0 months, all 18 follow-up patients survived without repeated surgeries. A follow-up computed tomography (CT) examination revealed a residual false lumen in the aortic arch in 3 patients and in the descending aorta in 8, with residual false lumen perfusion in one. Conclusions: Immediate recognition and surgical repair of ATAD that developed as a complication during cardiac surgical procedures are associated with low mortality and high intermediate-term survival.

Myectomy with and without Mitral Subvalvular Repair in Patients with Hypertrophic Obstructive Cardiomyopathy with Grade 3 to 4+ Mitral Regurgitation without Intrinsic Mitral Valve Disease: A Retrospective Observational Study.

Background: Hypertrophic obstructive cardiomyopathy (HOCM) with severe mitral regurgitation (MR) remains controversial for the choice of the concomitant mitral valve (MV) management versus septal myectomy alone. The impacts of different surgical strategies (concomitant mitral subvalvular procedures versus myectomy alone) on one-year results of surgical treatment of HOCM with grade 3 to 4+ MR without intrinsic MV disease were evaluated in this single-center, retrospective observational study. Methods: A total of 146 eligible patients were retrospectively screened into a combined group (n = 40) and an alone group (n = 106), depending on whether they underwent transaortic mitral subvalvular procedures. Perioperative outcomes were collected, and results at 1-year following surgery were compared. Results: Surgical mortality did not differ (0 for combined group vs. 0.9% for alone group, p = 0.538). Six patients (5.0% vs. 3.8%, p = 0.666) developed postoperative complete atrioventricular node block with permanent pacemaker implantation. No death or reoperation was recorded during a median follow-up of 18 months. At 1-year following surgery, (1) the provoked MR severity decreased from baseline in both groups with a significant difference between groups [1.0 (0-1.0) vs. 1.0 (1.0-1.3), p < 0.001]; (2) systolic anterior motion (SAM) was observed in 10 patients (0 vs. 10 in the alone group, p = 0.043); (3) the provoked gradient was also significantly lower than baseline value for each group, with a significant difference between the two groups (8.8 ± 4.3 mmHg vs. 12.1 ± 6.7 mmHg, p = 0.006); and (4) New York Heart Association class decreased from baseline value for each group (p < 0.001). Conclusions: In HOCM patients with grade 3 to 4+ MR without intrinsic MV disease, mitral subvalvular management during septal myectomy may be associated with a low incidence of SAM, improved MR, and a lower outflow tract gradient in comparison with septal myectomy alone.

Impact of Time in Motion on Blood Pressure Regulation Among Patients with Metabolic Syndrome.

PURPOSE OF REVIEW: This review assessed recent evidence on the association between objectively measured physical activity from wearable accelerometers and blood pressure (BP) in participants with metabolic syndrome (MetS). RECENT FINDINGS: Results directly related to BP were mixed, with some studies showing positive associations and others showing null results. Importantly, several studies noted that participants with MetS demonstrated greater improvements in components of MetS after engaging in higher amounts of daily physical activity. Although this suggests greater volume of physical activity may be a means to partially mitigate hypertension in those with MetS, it remains unclear whether physical activity or inactivity (i.e., sedentary behavior) is more strongly associated with MetS. Although there may be benefit to greater volumes of daily PA among hypertensive patients with MetS, more research is needed to quantify and define the amount of daily activity needed to improve health and refine clinical recommendations. Moreover, although the evidence for improving components of MetS through engaging in physical activity is high, the amount and type(s) of physical activity needed to achieve these benefits is unclear.

Multi-residue analysis of 206 pesticides in grass forage by the one-step quick, easy, cheap, effective, rugged, and safe method combined with ultrahigh-performance liquid chromatography quadrupole orbitrap mass spectrometry.

A novel method for detecting pesticide multi-residue in grass forage (alfalfa and oat) was established based on the one-step automatic extraction and purification technology of quick, easy, cheap, effective, rugged, and safe combined with ultrahigh-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry. The crushed sample was extracted with acetonitrile with 1% acetate, followed by a cleanup step with a primary-secondary amine, octadecylsilane, and graphitized carbon black. The extraction and purification were carried out using the one-step automatic pretreatment equipment. The target pesticides were acquired in positive ion electrospray ionization mode and full scan/data dependent secondary scan mode. The calibration curve shows good linearity over the corresponding concentration range, with the coefficient of determination greater than 0.99. The screening detection limits were 0.5-50 µg/kg, and the limit of quantification for the 206 pesticides was set at 1-50 µg/kg. At the spiking levels of one, two, and 10 times of limit of quantification, more than 95% of pesticides had recovery between 70-120%, with a relative standard deviation ≤20%. The method was proved to be simple, rapid, high-sensitivity, and could be routinely used for the high throughput screening and quantitative analysis of pesticide residues in alfalfa and oat.