Best Evidence Summary for the Prevention of Pressure Injuries in Orthopaedic Patients.

AIM: To systematically search, evaluate and synthesise the most robust evidence regarding pressure injury prevention in orthopaedic patients admitted to general wards. DESIGN: The present study provides an evidence-based summary of the most robust findings, adhering to the evidence guidelines established by the Center for Evidence-Based Nursing of Fudan University. METHOD: According to the "6S" model, a systematic search was conducted for literature on pressure injury prevention among orthopaedic patients in general wards. The types of literature included guidelines, clinical decisions, expert Consensus, evidence summaries, etc. The search period covered the time from the beginning of the database up to December 2023. DATA SOURCES: The following databases and resources were systematically searched: Up To Date, JBI, NICE, WOCN, NZWCS, etc. RESULTS: Fifteen literature sources were included, comprising one clinical decision, eight guidelines, one systematic review, and one expert Consensus. In these sources, a comprehensive collection of 34 pieces of best evidence was formed across six key topics: risk assessment, position management, skin care, device used for device-related pressure injury, nutritional assessment, and support, as well as health education and training. Among the evidence gathered, a strong recommendation was made for 18 pieces, while the remaining 16 received a weak recommendation. CONCLUSION: This study provides a comprehensive synthesis of the most robust evidence on pressure injury prevention in orthopaedic patients, encompassing 34 pieces of evidence that can serve as valuable references for clinical practice. Before implementing this evidence, it is crucial to evaluate the specific contextual factors within different countries and medical institutions, as well as the facilitators and barriers influencing its application by healthcare professionals and patient's preferences. Furthermore, targeted evidence selection should be conducted through careful screening and subsequent adjustments in implementation, thereby offering a more scientifically grounded basis for clinical nursing practice. Future research endeavours should prioritise investigating strategies for effective evidence utilisation. IMPLICATIONS FOR THE PROFESSION AND PATIENT CARE: The prevention of pressure injuries poses a significant challenge for orthopaedic patients. This study presents a synthesis of 34 pieces of best evidence to provide guidance on preventive measures for pressure injuries in orthopaedic patients. Adhering to and implementing these 34 pieces of evidence can effectively aid in preventing pressure injuries in clinical practice. This evidence encompasses risk assessment, position management, skin care, device usage for device-related pressure injuries, nutritional support and evaluation, and health education and training, establishing a comprehensive and systematic implementation process. Assessing the risk of pressure injuries during interventions serves as an essential prerequisite for developing effective strategies to prevent such injuries among orthopaedic patients. Ultimately, this study will offer valuable guidance to healthcare professionals worldwide regarding preventing pressure injuries in orthopaedic patients. IMPACT: Upon admission to the hospital, it is essential to conduct a risk assessment and implement evidence-based, individualised prevention measures for pressure ulcers in patients to prevent their occurrence. This study will provide valuable insights into preventing pressure injuries in orthopaedic patients admitted to orthopaedic wards for healthcare workers worldwide. STATE: The PRIMA manifest is utilised during the text preparation process. TRAIL REGISTRATION: ES20245365.

Selection and Validation of qRT-PCR Internal Reference Genes to Study Flower Color Formation in Camellia impressinervis.

Real-time quantitative PCR (qRT-PCR) is a pivotal technique for gene expression analysis. To ensure reliable and accurate results, the internal reference genes must exhibit stable expression across varied experimental conditions. Currently, no internal reference genes for Camellia impressinervis have been established. This study aimed to identify stable internal reference genes from eight candidates derived from different developmental stages of C. impressinervis flowers. We employed geNorm, NormFinder, and BestKeeper to evaluate the expression stability of these candidates, which was followed by a comprehensive stability analysis. The results indicated that CiTUB, a tubulin gene, exhibited the most stable expression among the eight reference gene candidates in the petals. Subsequently, CiTUB was utilized as an internal reference for the qRT-PCR analysis of six genes implicated in the petal pigment synthesis pathway of C. impressinervis. The qRT-PCR results were corroborated by transcriptome sequencing data, affirming the stability and suitability of CiTUB as a reference gene. This study marks the first identification of stable internal reference genes within the entire genome of C. impressinervis, establishing a foundation for future gene expression and functional studies. Identifying such stable reference genes is crucial for advancing molecular research on C. impressinervis.

The Euscaphis japonica genome and the evolution of malvids.

Malvids is one of the largest clades of rosids, includes 58 families and exhibits remarkable morphological and ecological diversity. Here, we report a high-quality chromosome-level genome assembly for Euscaphis japonica, an early-diverging species within malvids. Genome-based phylogenetic analysis suggests that the unstable phylogenetic position of E. japonica may result from incomplete lineage sorting and hybridization event during the diversification of the ancestral population of malvids. Euscaphis japonica experienced two polyploidization events: the ancient whole genome triplication event shared with most eudicots (commonly known as the γ event) and a more recent whole genome duplication event, unique to E. japonica. By resequencing 101 samples from 11 populations, we speculate that the temperature has led to the differentiation of the evergreen and deciduous of E. japonica and the completely different population histories of these two groups. In total, 1012 candidate positively selected genes in the evergreen were detected, some of which are involved in flower and fruit development. We found that reddening and dehiscence of the E. japonica pericarp and long fruit-hanging time promoted the reproduction of E. japonica populations, and revealed the expression patterns of genes related to fruit reddening, dehiscence and abscission. The key genes involved in pentacyclic triterpene synthesis in E. japonica were identified, and different expression patterns of these genes may contribute to pentacyclic triterpene diversification. Our work sheds light on the evolution of E. japonica and malvids, particularly on the diversification of E. japonica and the genetic basis for their fruit dehiscence and abscission.

Fungal Epithiodiketopiperazines Carrying α,ß-Polysulfide Bridges from Penicillium steckii YE, and Their Chemical Interconversion.

Some fungal epithiodiketopiperazine alkaloids display α,ß-polysulfide bridges alongside diverse structural variations. However, the logic of their chemical diversity has rarely been explored. Here, we report the identification of three new (2, 3, 8) and five known (1, 4-7) epithiodiketopiperazines of this subtype from a marine-derived Penicillium sp. The structure elucidation was supported by multiple spectroscopic analyses. Importantly, we observed multiple nonenzymatic interconversions of these analogues in aqueous solutions and organic solvents. Furthermore, the same biosynthetic origin of these compounds was supported by one mined gene cluster. The dominant analogue (1) demonstrated selective cytotoxicity to androgen-sensitive prostate cancer cells and HIF-depleted colorectal cells and mild antiaging activities, linking the bioactivity to oxidative stress. These results provide crucial insight into the formation of fungal epithiodiketopiperazines through chemical interconversions.

Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity.

Glycans possess significant chemical diversity; glycan binding proteins (GBPs) recognize specific glycans to translate their structures to functions in various physiological and pathological processes. Therefore, the discovery and characterization of novel GBPs and characterization of glycan-GBP interactions are significant to provide potential targets for therapeutic intervention of many diseases. Here, we report the biochemical, functional, and structural characterization of a 130-amino-acid protein, Y3, from the mushroom Coprinus comatus Biochemical studies of recombinant Y3 from a yeast expression system demonstrated the protein is a unique GBP. Additionally, we show that Y3 exhibits selective and potent cytotoxicity toward human T-cell leukemia Jurkat cells compared with a panel of cancer cell lines via inducing caspase-dependent apoptosis. Screening of a glycan array demonstrated GalNAcß1-4(Fucα1-3)GlcNAc (LDNF) as a specific Y3-binding ligand. To provide a structural basis for function, the crystal structure was solved to a resolution of 1.2 Å, revealing a single-domain αßα-sandwich motif. Two monomers were dimerized to form a large 10-stranded, antiparallel ß-sheet flanked by α-helices on each side, representing a unique oligomerization mode among GBPs. A large glycan binding pocket extends into the dimeric interface, and docking of LDNF identified key residues for glycan interactions. Disruption of residues predicted to be involved in LDNF/Y3 interactions resulted in the significant loss of binding to Jurkat T-cells and severely impaired their cytotoxicity. Collectively, these results demonstrate Y3 to be a GBP with selective cytotoxicity toward human T-cell leukemia cells and indicate its potential use in cancer diagnosis and treatment.

Genetics of Parkinson's disease and related disorders.

Parkinson's disease (PD) is a complex and heterogeneous neurological condition characterised mainly by bradykinesia, resting tremor, rigidity and postural instability, symptoms that together comprise the parkinsonian syndrome. Non-motor symptoms preceding and following clinical onset are also helpful diagnostic markers revealing a widespread and progressive pathology. Many other neurological conditions also include parkinsonism as primary or secondary symptom, confounding their diagnosis and treatment. Although overall disease course and end-stage pathological examination single out these conditions, the significant overlaps suggest that they are part of a continuous disease spectrum. Recent genetic discoveries support this idea because mutations in a few genes (α-synuclein, LRRK2, tau) can cause partially overlapping pathologies. Additionally, mutations in causative genes and environmental toxins identify protein homeostasis and the mitochondria as key mediators of degeneration of dopaminergic circuits in the basal ganglia. The evolving mechanistic insight into the pathophysiology of PD and related conditions will contribute to the development of targeted and effective symptomatic treatments into disease-modifying therapies that will reduce the burden of these dreadful conditions.

High-Yield Production of Herbicidal Thaxtomins and Thaxtomin Analogs in a Nonpathogenic Streptomyces Strain.

Thaxtomins are virulence factors of most plant-pathogenic Streptomyces strains. Due to their potent herbicidal activity, attractive environmental compatibility, and inherent biodegradability, thaxtomins are key active ingredients of bioherbicides approved by the U.S. Environmental Protection Agency. However, the low yield of thaxtomins in native Streptomyces producers limits their wide agricultural applications. Here, we describe the high-yield production of thaxtomins in a heterologous host. The thaxtomin gene cluster from S. scabiei 87.22 was cloned and expressed in S. albus J1074 after chromosomal integration. The production of thaxtomins and nitrotryptophan analogs was observed using liquid chromatography-mass spectrometry (LC-MS) analysis. When the engineered S. albus J1074 was cultured in the minimal medium Thx defined medium supplemented with 1% cellobiose (TDMc), the yield of the most abundant and herbicidal analog, thaxtomin A, was 10 times higher than that in S. scabiei 87.22, and optimization of the medium resulted in the highest yield of thaxtomin analogs at about 222 mg/liter. Further engineering of the thaxtomin biosynthetic gene cluster through gene deletion led to the production of multiple biosynthetic intermediates important to the chemical synthesis of new analogs. Additionally, the versatility of the thaxtomin biosynthetic system in S. albus J1074 was capitalized on to produce one unnatural fluorinated analog, 5-fluoro-thaxtomin A (5-F-thaxtomin A), whose structure was elucidated by a combination of MS and one-dimensional (1D) and 2D nuclear magnetic resonance (NMR) analyses. Natural and unnatural thaxtomins demonstrated potent herbicidal activity in radish seedling assays. These results indicated that S. albus J1074 has the potential to produce thaxtomins and analogs thereof with high yield, fostering their agricultural applications.IMPORTANCE Thaxtomins are agriculturally valuable herbicidal natural products, but the productivity of native producers is limiting. Heterologous expression of the thaxtomin gene cluster in S. albus J1074 resulted in the highest yield of thaxtomins ever reported, representing a significant leap forward in its wide agricultural use. Furthermore, current synthetic routes to thaxtomins and analogs are lengthy, and two thaxtomin biosynthetic intermediates produced at high yields in this work can provide precursors and building blocks to advanced synthetic routes. Importantly, the production of 5-F-thaxtomin A in engineered S. albus J1074 demonstrated a viable alternative to chemical methods in the synthesis of new thaxtomin analogs. Moreover, our work presents an attractive synthetic biology strategy to improve the supply of herbicidal thaxtomins, likely finding general applications in the discovery and production of many other bioactive natural products.

The peripheral blood of Aß binding RBC as a biomarker for diagnosis of Alzheimer's disease.

BACKGROUND: in vitro, it has been reported that amyloid ß (Aß) is bound to red blood cells (RBCs) and this process damages the red cell. Also, a possible relationship between RBCs and Alzheimer's disease (AD) is supported by the findings of RBC impairment in AD. Therefore, Aß fibrils bounding RBC are of great interest as potential biomarkers. METHODS: in this study, we focused on Aß amyloid fibrils and/or aggregation on the peripheral RBC from 50 subjects with AD and 50 healthy controls (HCs) through thioflavin T (ThT) staining followed by immunofluorescence assay to confirm the presence of Aß amyloid fibrils and/or aggregation on the RBC. Then we optimised fluorescence staining and imaging conditions and analysed the images obtained by image processing software. RESULTS: we have analysed RBC morphology in blood from 50 subjects with AD and 50 HCs found that 16.8% of the RBCs are elongated as compared with 6.7% in normal controls (P < 0.01), and there is a negative correlation between the two parameters (P < 0.05). Our study showed that 98% of AD peripheral RBCs were amyloid binding-positive (ranging from 2 to 30%), while only 38% that of RBCs (ranging from 2 to 3.4%) were in HCs. We also found four modified morphologies of RBCs triggered by Aß binding, which may serve as an ancillary investigation and indicate the progression of AD. CONCLUSION: we first directly prove the existence of Aß binding RBCs in peripheral blood. In addition, we observed new modified morphologies of RBC triggered by Aß binding, all of those can serve as a biomarker for the diagnosis and progression of AD.

Research on the Method of Detecting TPN-Labeled Tumor Cells in Pleural Effusion Based on the Microfluidic Chip.

The clinical diagnosis of a malignant pleural effusion (MPE) is still based on the detection of tumor cells in the pleural effusion. The question of how to improve the efficiency and accuracy of detecting an MPE still remains. This study explores the use of microfluidic technology to concentrate cells in an MPE and achieved the detection of the cell marker TPN in the microarray capture area. TPN is a mitochondria-specific bio-probe that can identify tumor cells on the basis of differences in the mitochondrial potential. First, we designed a microfluidic chip to analyze its performance. The results show that when the total flow rate of the injected chip was 12 mL/h and the volume ratio of cell separation liquid to cell suspension was 1:1, the target cells (A549, MCF-7, and Hela) were enriched and the purity was improved to 98.7-99.3%. Finally, an MPE from cancer patients was used to detect the chip's ability to isolate and enrich tumor cells. Furthermore, the fluorescent identification of the TPN within the tumor cells was simultaneously achieved on the microfluidic chip. In conclusion, the potential to improve the efficiency of the clinical diagnosis of MPEs is provided by the chip structure and analysis conditions explored in this study.

Efficacy and safety of short-term high dosage dual antiplatelet therapy after 0.6 mg/kg rt-PA intravenous thrombolysis for acute ischemic stroke.

OBJECTIVE: To evaluate the efficacy and safety of short-term high-dose of dual antiplatelet therapy after 0.6 mg/kg rt-PA intravenous thrombolysis for acute ischemic stroke (AIS). METHODS: All 208 patients with AIS were randomized into group 1 (103 cases, after 0.6 mg/kg rt-PA, 300 mg of oral aspirin(ASP) q.d. and 225 mg of oral clopidogrel (CLO) q.d. for for 5 days, then 100 mg of oral ASP q.d. for the next 85 days and 75 mg of oral CLO q.d. for the next 16 days) and group 2 (105 cases, after 0.9 mg/kg rt-PA, 100 mg of oral ASP q.d. for 90 days and 75 mg of oral CLO q.d. for 21 days).The efficacy index was the mRS score, NIHSS score and recurrence risk of stroke, while the safety index was the incidence of bleeding events and mortality. All parameters were evaluated at 30 and 90 days after thrombolysis. Patients whose characteristics may provide the best treatment benefit were further analyzed using the logistic regression model in group 1. RESULTS: The proportion of mRS scores between 0 and 1 in group 1 was higher than that in group 2 at both 30 days (44.7% vs 32.4%, P < .05) and 90 days (50.5% vs 35.2%, P < .05). Compared to group 2, the proportion of NIHSS scores less than 4 was significantly higher in group 1 at both 30 days (37.9% vs 25.7%, P < .05) and 90 days (46.6% vs 30.5%, P < .05). At 90 days, Group 1 had a lower stroke recurrence risk than Group 2 (3.9% vs 10.5%, P < .05). The incidence of SICH was significantly different between the 2 groups at both 30 days (2.9% vs 9.5%, P < .05) and 90 days (2.9% vs 10.5%, P < .05). However, other bleeding events and mortality rates were not significantly different between the 2 groups. The lower the baseline NIHSS score and the shorter the OTT, the more favorable the outcomes obtained at 90 days. CONCLUSIONS: Compared to standard doses, short term high-dose dual antiplatelet therapy after 0.6 mg/kg rt-PA intravenous thrombolysis may be a good choice for AIS patients.

Correlation Between Blood Glucose Variability and Early Therapeutic Effects After Intravenous Thrombolysis With Alteplase and Levels of Serum Inflammatory Factors in Patients With Acute Ischemic Stroke.

Objective: To investigate the effects of blood glucose variability on early therapeutic effects after intravenous thrombolysis with alteplase and the levels of serum inflammatory factors in patients with acute ischemic stroke (AIS). Methods: We enrolled AIS patients who received intravenous thrombolysis within 4.5 h of the onset of symptoms. Clinical data, including the National Institutes of Health Stroke Scale (NIHSS), glycosylated hemoglobin, mean blood glucose, standard deviation of blood glucose, mean amplitude of glycemic excursion, mean variation coefficient of blood glucose, interleukin-6 (IL-6), active matrix metalloproteinase-9 (MMP-9), tumor necrosis factor α (TNF-α), and hypersensitive C-reactive protein (hs-CRP) levels, were compared between a group who showed improvement (the improvement group) and a group who did not show improvement (the non-improvement group). Relevant factors for early neurological improvement after thrombolysis with alteplase were analyzed by using multivariate logistic regression models. A Pearson linear correlation analysis was also performed on blood glucose variation and inflammatory factor levels within the two groups. Results: A total of 146 patients were included, 63 of which had early symptom improvement (43.15%). The diabetes ratio, atrial fibrillation ratio, baseline NHISS score, random blood glucose at admission, and glycosylated hemoglobin of patients in the improvement group were significantly lower than those in the non-improvement group (P < 0.05 in all cases). The mean blood glucose, standard deviation of blood glucose, mean amplitude of glycemic excursion, and mean blood glucose variation coefficients of patients in the improvement group were significantly lower than those in the non-improvement group (P < 0.05). Serum inflammatory factor levels, including IL-6, MMP-9, TNF-α and hs-CRP, were significantly lower in patients in the improvement group compared to patients in the non-improvement group (P < 0.05). Multivariate logistic regression analysis showed that baseline NIHSS scores (OR = 1.28, 95% CI = 1.05-1.62, P = 0.02), glycosylated hemoglobin scores (OR = 2.57, 95% CI = 1.78-3.98, P = 0.0005), diabetes (OR = 13.10, 95% CI = 1.63~131.45, P = 0.021), the mean amplitude of glycemic excursion (OR = 2.98, 95% CI = 1.92-5.00, P < 0.0001), and the mean variation coefficient of blood glucose (OR = 1.40, 95% CI = 1.26-1.60, P = 0.0078) were significantly correlated with early symptom improvement after thrombolysis. Pearson linear correlation analysis showed that the standard deviation of blood glucose, mean amplitude of glycemic excursion, and the mean variation coefficient of blood glucose were significantly positively correlated with IL-6, MMP-9, TNF-α and hs-CRP levels (P < 0.01). Conclusions: Blood glucose variability is correlated with early neurological improvement after intravenous thrombolysis with alteplase in AIS patients. With the increase of blood glucose fluctuation range, the inflammatory response is enhanced, which affects the prognosis of patients.

Rates of intravenous thrombolysis and endovascular therapy for acute ischaemic stroke in China between 2019 and 2020.

BACKGROUND: In recent years, a series of initiatives have been launched to promote intravenous thrombolysis (IVT) and endovascular therapy (EVT) for acute ischaemic stroke (AIS) in China. We aimed to update the rates of IVT and EVT in China between 2019 and 2020 and to evaluate the current IVT and EVT according to hospital grades. METHODS: Cross-sectional data on patients receiving IVT/EVT were derived from the Bigdata Observatory platform for Stroke of China (BOSC). The monthly number of discharged patients with a principal diagnosis of AIS was derived from the first pages of medical records of each hospital. The rates and information of IVT and EVT were analysed according to hospital grades. FINDINGS: During this period, 938 tertiary hospitals and 786 secondary hospitals from 31 provinces continuously reported data to the BOSC. The overall IVT rate for AIS was 5·64%, and the EVT rate was 1·45%. The IVT rate in secondary hospitals was higher than that in tertiary hospitals (6·39% vs. 5·39%, P < 0·001), whereas the EVT rate in secondary hospitals was much lower than that in tertiary hospitals (0·29% vs. 1·84%, P < 0·001). Significant differences in demographic and clinical characteristics of patients receiving IVT/EVT were also shown between tertiary and secondary hospitals. INTERPRETATION: The rates of IVT and EVT for AIS have greatly increased in China, but there is still a large gap compared with developed countries. Hospital inhomogeneity in IVT and EVT suggests the importance of developing a region-specific network for stroke treatment. FUNDING: None.

miR-378a-5p regulates CAMKK2/AMPK pathway to contribute to cerebral ischemia/reperfusion injury-induced neuronal apoptosis.

INTRODUCTION: The pathological mechanism of cerebral ischemia/reperfusion (CIR) injury is complicated and unclear. Apart from the involvement of many low-molecular factors it was found that several miRNAs were dysregulated during and after CIR injury in cell models. This study aimed to explore the effects of miR-378a-5p on in vitro model of (CIR) injury-induced neuronal apoptosis and provide a new mechanism of CIR injury. MATERIAL AND METHODS: Primary hippocampal neurons were isolated from newborn Sprague-Dawley rats. Oxygen- glucose deprivation/reoxygenation (OGDR) for 24 h and 48 h was used as an in vitro model of CIR. Cell viability was measured using MTT assay and apoptosis was determined by flow cytometry. Quantitative real time PCR (qRT-PCR) assay and Western blotting were used to examine mRNA and protein expressions, respectively. TargetScan was used to predict the direct target of miR-378a-5p and luciferase assay was used to validate that calmodulin-dependent protein kinase kinase-2 (CAMKK2) was the direct target of miR-378a-5p. RESULTS: miR-378a-5p expression was significantly increased after OGDR at 24 h and 48 h. After OGDR, cell viability was reduced, which was reversed by miR-378a-5p and enhanced by shCAMKK2 plasmid. Cell apoptosis was increased after OGDR, which was prevented by miR-378a-5p and enhanced by shCAMKK2 plasmid. Results of TargetScan and luciferase assay demonstrated that miR-378a-5p could directly bind to 3'-untranslated region (3'-UTR) of CAMKK2. Both mRNA and protein expression of CAMKK2 were downregulated by miR-378a-5p mimics and upregulated by miR-378a-5p inhibitors. Phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) was positively associated with expression of CAMKK2. CONCLUSIONS: Data of this study indicated that miR-378a-5p was significantly overexpressed after OGDR. miR-378a-5p could bind to 3'-UTR of CAMKK2 to inhibit cell proliferation through regulation of CAMKK2/AMPK pathway providing a new mechanism and biomarker for the diagnosis and potential treatment of CIR injury.

Genkwanin suppresses MPP+-induced cytotoxicity by inhibiting TLR4/MyD88/NLRP3 inflammasome pathway in a cellular model of Parkinson's disease.

Parkinson's disease (PD) is a complicated multifactorial neurodegenerative disorder. Oxidative stress, neuroinflammatory response, and activation of apoptosis have been proposed to be tightly involved in the pathogenesis of PD. Genkwanin is a typical bioactive non-glycosylated flavonoid with anti-inflammatory and anti-oxidant activities. However, the effect of genkwanin on PD remains unclear. Cell viability, lactate dehydrogenase (LDH) release, caspase-3/7 activity, and apoptosis was evaluated by MTT, LDH release assay, caspase-3/7 activity assay, and TUNEL assay, respectively. The secretion of prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6 were measured by respective commercial ELISA kits. The mRNA expression of TNF-α, IL-1ß, and IL-6 was detected by qRT-PCR. The protein levels of cycloxygenase-2 (COX-2), toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and NOD-like receptor (NLR) protein: 3 (NLRP3) were determined by western blot analysis. Genkwanin at concentrations less than 40 µM had no impact on cell viability and LDH release. Genkwanin suppressed MPP+-induced neuroinflammation in SH-SY5Y cells. MPP+ treatment inhibited cell viability, increased LDH release, apoptosis, and ROS generation, and reduced superoxide dismutase (SOD) activity in SH-SY5Y cells, which were abolished by genkwanin treatment. Genkwanin suppressed MPP+-induced activation of TLR4/MyD88/NLRP3 inflammasome pathway in SH-SY5Y cells. TLR4 overexpression weakened the anti-inflammatory and anti-neurotoxicity of genkwanin in SH-SY5Y cells. In conclusion, genkwanin attenuated neuroinflammation and neurotoxicity by inhibiting TLR4/MyD88/NLRP3 inflammasome pathway in MPP+-induced cellular model of PD.

GLP-1 Induces the Expression of FNDC5 Derivatives That Execute Lipolytic Actions.

Multiple GLP-1-derived therapeutics are clinically used to treat type 2 diabetes and obesity. However, the underlying mechanism of how these drugs regulate the body weight of obese patients remains incompletely understood. Here, we report that the lipolysis effects of GLP-1 on ß cells can depend on its induced expression of fibronectin type III domain containing 5 (FNDC5). The transmembrane FNDC5 is a precursor of the recently identified hormone irisin that possesses a range of bioactivities, including anti-obesity and anti-diabetes. We revealed that GLP-1 upregulates the expression and secretion of FNDC5 in ß cells, while GLP-1 itself fails to activate the lipolysis genes in FNDC5-knockout ß cells. In addition, liraglutide, a clinically used GLP-1 receptor agonist, induced the expression of FNDC5 in mouse pancreas and brain tissues and increased the serum level of secreted FNDC5. Furthermore, we observed the expression of the well-known membrane-associated FNDC5 and a novel, secretable FNDC5 (sFNDC5) isoform in ß cells and multiple rat tissues. Recombinant sFNDC5 stimulated lipolysis of wild type and FNDC5-knockout ß cells. This new isoform further induced lipolysis and browning of adipocytes, and similar to irisin, executed potent anti-obesity activities in an obese mouse model. Overall, our studies provided new mechanistic insights into GLP-1's anti-obesity actions in which GLP-1 induces the secretion of FNDC5 derivatives from its responsive organs that then mediate its anti-obesity activities.

A Modular Synthetic Route Involving N-Aryl-2-nitrosoaniline Intermediates Leads to a New Series of 3-Substituted Halogenated Phenazine Antibacterial Agents.

Pathogenic bacteria demonstrate incredible abilities to evade conventional antibiotics through the development of resistance and formation of dormant, surface-attached biofilms. Therefore, agents that target and eradicate planktonic and biofilm bacteria are of significant interest. We explored a new series of halogenated phenazines (HP) through the use of N-aryl-2-nitrosoaniline synthetic intermediates that enabled functionalization of the 3-position of this scaffold. Several HPs demonstrated potent antibacterial and biofilm-killing activities (e.g., HP 29, against methicillin-resistant Staphylococcus aureus: MIC = 0.075 µM; MBEC = 2.35 µM), and transcriptional analysis revealed that HPs 3, 28, and 29 induce rapid iron starvation in MRSA biofilms. Several HPs demonstrated excellent activities against Mycobacterium tuberculosis (HP 34, MIC = 0.80 µM against CDC1551). This work established new SAR insights, and HP 29 demonstrated efficacy in dorsal wound infection models in mice. Encouraged by these findings, we believe that HPs could lead to significant advances in the treatment of challenging infections.

Cyanobacterial Dihydroxyacid Dehydratases Are a Promising Growth Inhibition Target.

Microbes are essential to the global ecosystem, but undesirable microbial growth causes issues ranging from food spoilage and infectious diseases to harmful cyanobacterial blooms. The use of chemicals to control microbial growth has achieved significant success, while specific roles for a majority of essential genes in growth control remain unexplored. Here, we show the growth inhibition of cyanobacterial species by targeting an essential enzyme for the biosynthesis of branched-chain amino acids. Specifically, we report the biochemical, genetic, and structural characterization of dihydroxyacid dehydratase from the model cyanobacterium Synechocystis sp. PCC 6803 (SnDHAD). Our studies suggest that SnDHAD is an oxygen-stable enzyme containing a [2Fe-2S] cluster. Furthermore, we demonstrate that SnDHAD is selectively inhibited in vitro and in vivo by the natural product aspterric acid, which also inhibits the growth of representative bloom-forming Microcystis and Anabaena strains but has minimal effects on microbial pathogens with [4Fe-4S] containing DHADs. This study suggests DHADs as a promising target for the precise growth control of microbes and highlights the exploration of other untargeted essential genes for microbial management.

Rapid kill assessment of an N-arylated NH125 analogue against drug-resistant microorganisms.

While a number of disinfection techniques are employed in healthcare units, the eradication of drug-resistant microorganisms remains a challenge. We recently reported N-arylated NH125 analogue 1, which demonstrated potent biofilm eradication and antibacterial activities against a panel of drug-resistant pathogens. The broad-spectrum activities observed for 1 along with its rapid eradication of MRSA persister cells suggested that this agent, and related analogues, can serve as disinfectants for antibiotic resistant pathogens in healthcare settings. Here, we report the rapid bactericidal activities of 1 against a panel of exponentially-growing, drug-resistant pathogens. Against MRSA, MRSE, VRE and MDR A. baumannii, 1 eradicated bacterial cells after five minutes when tested at 50 µM (3- to 6-log reduction of CFU per mL). We highlighted the rapid killing activities by demonstrating that 1 eradicates 99.99% of viable MRSA 1707 cells in one minute (50 µM, 4-log reduction of CFU per mL). In addition, 1 rapidly eradicated fungal pathogen C. neoformans in kill kinetic experiments. A solution of 1 demonstrated similar shelf stability to known disinfectant BAC-16 when tested up to 111 days after being stored. Collectively, our data highlights the potential of 1 to be used as a disinfecting agent to prevent healthcare-associated, drug-resistant infections.

The efficacy of homemade tolcapone in the treatment of patients with Parkinsons disease.

The aim of the study was to investigate the efficacy of homemade tolcapone in treatment of patients with Parkinson's disease (PD). Eighty patients with PD were subjected to randomized, double-blind, placebo-controlled and parallel-group study. PD patients were randomly divided into the tolcapone treatment group (41 cases) and placebo group (39 cases). Each patient received 1 pill of tolcapone or placebo, 3 times per day for 26 weeks. Efficacy was evaluated using the internationally used unified Parkinson's disease rating scale (UPDRS) scoring system for PD symptoms. After the treatment for 26 weeks, the cognitive function, tremor, muscle stiffness, voluntary movement and autonomic nerve symptoms were compared between the two groups using UPDRS scores. Compared with the placebo group, cognitive function, muscle stiffness and voluntary movement reduction were significantly improved in patients of the tolcapone group (P<0.05). However, no significant differences in UPDRS scores of autonomic nerve symptoms and tremor were found between two groups after treatment (P>0.05). Tolcapone, a catechol-O-methyl transferase (COMT) inhibitor, can improve the motor function of patients with PD, especially exercise and muscle stiffness. Tolcapone can also improve the cognitive function of patients.