A facile fluorescence-coupling approach to visualizing leonurine uptake and distribution in living cells and Caenorhabditis elegans.

BACKGROUND: Caenorhabditis elegans (C. elegans) has been recognized for being a useful model organism in small-molecule drug screens and drug efficacy investigation. However, there remain bottlenecks in evaluating such processes as drug uptake and distribution due to a lack of appropriate chemical tools. PURPOSE: This study aims to prepare fluorescence-labeled leonurine as an example to monitor drug uptake and distribution of small molecule in C. elegans and living cells. METHODS: FITC-conjugated leonurine (leonurine-P) was synthesized and characterized by LC/MS, NMR, UV absorption and fluorescence intensity. Leonurine-P was used to stain C. elegans and various mammalian cell lines. Different concentrations of leonurine were tested in conjunction with a competing parent molecule to determine whether leonurine-P and leonurine shared the same biological targets. Drug distribution was analyzed by imaging. Fluorometry in microplates and flow cytometry were performed for quantitative measurements of drug uptake. RESULTS: The UV absorption peak of leonurine-P was 490∼495 nm and emission peak was 520 nm. Leonurine-P specifically bound to endogenous protein targets in C. elegans and mammalian cells, which was competitively blocked by leonurine. The highest enrichment levels of leonurine-P were observed around 72 h following exposure in C. elegans. Leonurine-P can be used in a variety of cells to observe drug distribution dynamics. Flow cytometry of stained cells can be facilely carried out to quantitatively detect probe signals. CONCLUSIONS: The strategy of fluorescein-labeled drugs reported herein allows quantification of drug enrichment and visualization of drug distribution, thus illustrates a convenient approach to study phytodrugs in pharmacological contexts.

First identification of multidrug-resistant Acinetobacter bereziniae isolates harboring bla NDM-1 from hospitals in South China.

This study is a first report on the identification of multidrug-resistant (MDR) Acinetobacter bereziniae among non-baumannii acinetobacters that had previously escaped automated laboratory detection, and characterize their clinical courses of infection at two tertiary-care hospitals in Shenzhen city, China (2015-2017). Herein, definitive identification by PCR was performed with universal and species-specific primers targeting 16S rDNA and rpoB genes, respectively, followed by Sanger sequencing and blast analysis. Antimicrobial susceptibility of A. bereziniae isolates was assessed accordingly. Three of the five identified A. bereziniae isolates exhibited carbapenem-resistance and were subjected to a multiplex PCR assay to detect drug-resistance genes. Sequences of the rpoB amplicon were aligned with curated sequences from global databases for phylogenetic analysis on evolutionary relations. Five clinical isolates of A. bereziniae were thereby re-identified, whose infections were primarily nosocomial. Automated identification and susceptibility testing systems (Phoenix-100 and VITEK 2) proved insufficient for discriminating A. bereziniae from other acinetobacters such as Acinetobacter baumannii and Acinetobacter guillouiae. Among these isolates, three exhibited carbapenem-resistant phenotypes indistinguishable from that of carbapenem-resistant A. baumannii. The carbapenem-resistant A. bereziniae isolates were subsequently confirmed to carry a bla NDM-1 (New Delhi metallo-ß-lactamase-1) gene downstream of ISAba125. Phylogenetic analysis revealed that A. bereziniae isolates evolved slowly but independently in local habitats. A. bereziniae isolates are difficult to distinguish by traditional automated detection systems. PCR-based identification via amplification and sequencing of selected house-keeping genes provides sufficient resolution for discriminating the isolates.

Association between screen time and hyperactive behaviors in children under 3 years in China.

Background: Screen time during early life has increased dramatically among Chinese children. Excessive screen time has raised growing concerns about the neuropsychological development of children. The effects of screen exposure on early life and the boundary between screen time and hyperactive behaviors are well worth investigating. We examined associations between screen time and hyperactive behaviors in children under the age of 3 years using data from the Longhua Children Cohort Study (LCCS). Methods: A cross-sectional study was conducted among 42,841 3-year-old children from Longhua District, Shenzhen. Information on socio-demographic characteristics, children's annual screen time since birth, and hyperactive behaviors (measured by the Conners Parental Symptom Questionnaire) was collected through self-administered structured questionnaires completed by the primary caregiver. A series of logistic regression models assessed the association between screen time and hyperactive behaviors. Results: The average daily screen time of children under the age of 3 years was 55.83 ± 58.54 min, and screen time increased with age. Binomial logistic regression analysis found that the earlier the screen exposure, the greater the risk of hyperactive behaviors. Using binary logistic regression model, after controlling for confounding factors, the study found that more screen time was more associated with hyperactive behaviors. For children aged 0-3 years with daily screen time exceeding 90, 120, 150, and 180 min, the risk values for hyperactive behaviors were 1.98 [95% confidence interval (CI): 1.05, 3.78), 2.71 (95%CI:1.38, 5.30), 3.17 (95% CI: 1.50, 6.65), and 4.62 (95% CI: 2.45, 8.71)], respectively. Conclusion: Early screen exposure may be associated with hyperactive behaviors in children under the age of 3 years. More than 90 min of screen time per day in children under 3 years was associated with hyperactive behaviors. The findings support the importance of screen time interventions for children under 3 years.

Daptomycin and linezolid for severe methicillin-resistant Staphylococcus aureus psoas abscess and bacteremia: A case report and review of the literature.

BACKGROUND: Vancomycin remains a first-line treatment drug as per the treatment guidelines for methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. However, a number of gram-positive cocci have developed resistance to several drugs, including glycopeptides. Therefore, there is an urgent need for effective and innovative antibacterial drugs to treat patients with infections caused by drug-resistant bacteria. CASE SUMMARY: A 24-year-old male was admitted to hospital owing to lumbago, fever, and hematuria. Computed tomography (CT) results showed an abscess in the psoas major muscle of the patient. Repeated abscess drainage and blood culture suggested MRSA, and vancomycin was initiated. However, after day 10, CT scans showed abscesses in the lungs and legs of the patient. Therefore, treatment was switched to daptomycin. Linezolid was also added considering inflammation in the lungs. After 10 d of the dual-drug anti-MRSA treatment, culture of the abscess drainage turned negative for MRSA. On day 28, the patient was discharged without any complications. CONCLUSION: This case indicates that daptomycin combined with linezolid is an effective remedy for bacteremia caused by MRSA with pulmonary complications.

High molecular weight fibroblast growth factor 2 induces apoptosis by interacting with complement component 1 Q subcomponent-binding protein in vitro.

Fibroblast growth factor 2 (FGF2) is a multifunctional cell growth factor that regulates cell proliferation, differentiation, adhesion, migration, and apoptosis. FGF2 has multiple isoforms, including an 18-kDa low molecular weight isoform (lo-FGF2) and 22-, 23-, 24-, and 34-kDa high molecular weight isoforms (hi-FGF2). Hi-FGF2 overexpression induces chromatin compaction, which requires the mitochondria and leads to apoptosis. Complement component 1 Q subcomponent-binding protein (C1QBP) plays an important role in mitochondria-dependent apoptosis by regulating the opening of the mitochondrial permeability transition pore. However, the interaction between C1QBP and hi-FGF2 and its role in hi-FGF2-mediated apoptosis remain unclear. Here, we found that hi-FGF2 overexpression induced depolarization of the mitochondrial membrane, cytochrome c release into the cytosol, and a considerable increase in C1QBP messenger RNA and protein expression. Furthermore, coimmunoprecipitation results showed that the mitochondrial protein, C1QBP, interacts with hi-FGF2. C1QBP knockdown using small interfering RNA significantly decreased the localization of hi-FGF2 to the mitochondria and increased the rate of apoptosis. Our results highlight a novel mechanism underlying hi-FGF2-induced, mitochondria-driven cell death involving the direct interaction between hi-FGF2 and C1QBP and the upregulation of C1QBP expression.