Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Regenerative Braking Algorithm for Parallel Hydraulic Hybrid Vehicles Based on Fuzzy Q-Learning

The use of regenerative braking systems is an important approach for improving the travel mileage of electric vehicles, and the use of an auxiliary hydraulic braking energy recovery system can improve the efficiency of the braking energy recovery process. In this paper, we present an algorithm for optimizing the energy recovery efficiency of a hydraulic regenerative braking system (HRBS) based on fuzzy Q-Learning (FQL). First, we built a test bench, which was used to verify the accuracy of the hydraulic regenerative braking simulation model. Second, we combined the HRBS with the electric vehicle in ADVISOR. Third, we modified the regenerative braking control strategy by introducing the FQL algorithm and comparing it with a fuzzy-control-based energy recovery strategy. The simulation results showed that the power savings of the vehicle optimized by the FQL algorithm were improved by about 9.62% and 8.91% after 1015 cycles and under urban dynamometer driving schedule (UDDS) cycle conditions compared with a vehicle based on fuzzy control and the dynamic programming (DP) algorithm. The regenerative braking control strategy optimized by the fuzzy reinforcement learning method is more efficient in terms of energy recovery than the fuzzy control strategy

Interleukin-1β Promotes Epithelial-Derived Alveolar Elastogenesis via αvβ6 Integrin-Dependent TGF-β Activation

Background/Aims: IL-1β creates persistent pulmonary inflammation accompanied by elevated transforming growth factor β (TGF-β levels and is associated with abnormal elastogenesis, which is observed in bronchopulmonary dysplasia (BPD). Although progress has been made in this field, the mechanisms underlying this process remain only partially understood. Methods: We assessed aberrant elastin localization-associated signaling in mouse pups exposed to 85% O2 treated with either IL-1Ra or 1D11, using morphometric analyses, quantitative RT-PCR, immunostaining, and ELISA. We also evaluated the derivation of elastin-producing cells using dual marker tracking. The regulatory mechanisms of IL-1β were investigated in vitro in lung epithelial and mesenchymal cells. Results: Elevated levels of IL-1β, αvβ6 and TGF-β1 were each associated with aberrant elastin production in O2-exposed lungs. IL-1Ra abolished TGF-β1 activation and αvβ6 upregulation, which occurred as a result of exposure to hyperoxia, whereas 1D11 had no discernible effect on the expression of either αvβ6 or IL-1β even following O2-exposure, suggesting that IL-1β was initially induced. Additionally, double staining revealed the presence of epithelium-derived elastin-producing cells, which was confirmed via in vitro IL-1β stress-induced epithelial-mesenchymal transformation (EMT) morphological and molecular marker changes, which may explain the altered lung elastin deposition and defective septation observed in BPD. Conclusions: These data support the hypothesis that IL-1β was initially induced by hyperoxia; αvβ6 subsequently interacted with and activated TGF-β1, acting as an epithelial/mesenchymal signaling molecule that contributed to excessive alveolar elastogenesis, the primary pathological feature of BPD

Diverse Synthesis of Triarylselenonium Salts and <i>o</i>‑(Alkoxy)aryl Aryl Selanes via Insertion of Benzyne into the SeO Bond

An insertion of benzyne into the SeO bond has been realized. In this reaction, two types of compounds containing selenium could be synthesized from different substrates. When diaryl selenium oxides were used, triarylselenonium salts were furnished, while o-(alkoxy)phenyl phenyl selanes could be produced from aryl alkyl selenium oxides

A long non-coding RNA HOTTIP expression is associated with disease progression and predicts outcome in small cell lung cancer patients

Abstract Background Despite progress in treatment of small cell lung cancer (SCLC), the biology of the tumor still remains poorly understood. Recently, we globally investigated the contributions of lncRNA in SCLC with a special focus on sponge regulatory network. Here we report lncRNA HOTTIP, which is specifically amplified in SCLC, is associated with SCLC proliferation and poor prognosis of patients. Methods RT-qPCR was used to investigate the expression of HOTTIP in SCLC tissues and cell lines. The role of HOTTIP in SCLC cell proliferation was demonstrated by CCK8 assay, colony formation assay, flow cytometry analysis and in vivo SCLC xenograft model in nude mice through HOTTIP loss- and gain-of-function effects. Western blot assay was used to evaluate gene expression in cell lines at protein level. RNA pull-down, Mass spectrometry and RNA binding protein immunoprecipitation (RIP) were performed to confirm the molecular mechanism of HOTTIP involved in SCLC progression. Results We found that HOTTIP was overexpressed in SCLC tissues, and its expression was correlated with the clinical stage and the shorter survival time of SCLC patients. Moreover, HOTTIP knockdown could impair cell proliferation, affect the cell cycle and inhibit tumor growth of mice, while HOTTIP overexpression might enhance cell proliferation and cell cycle in vitro and in vivo. Mechanistic investigations showed that HOTTIP functions as an oncogene in SCLC progression by sponging miR-574-5p and affecting the expression of polycomb group protein EZH1. Conclusions Overall, we identified that HOTTIP was involved in SCLC tumorigenesis through the ceRNA network “HOTTIP/miR-574-5p/EZH1”. Our findings not only illuminate how HOTTIP confers an oncogenic function in SCLC pathogenesis, but also underscore a novel gene expression governing hallmarks in the disease

Additional file 4: of A long non-coding RNA HOTTIP expression is associated with disease progression and predicts outcome in small cell lung cancer patients

Legends of supplementary figures, primers, RNAi sequences, miRNA and antibodies information. (DOC 633 kb

Additional file 1: of A long non-coding RNA HOTTIP expression is associated with disease progression and predicts outcome in small cell lung cancer patients

HOTTIP and miR-574-5p were screened out by microarray and RT-qPCR methods. (JPEG 275 kb

Additional file 2: of A long non-coding RNA HOTTIP expression is associated with disease progression and predicts outcome in small cell lung cancer patients

HOTTIP regulated the cell cycle of SCLC cells. (JPEG 353 kb

Acute traumatic coma awakening by right median nerve electrical stimulation: a randomised controlled trial.

PURPOSE: Severe traumatic brain injury (TBI) leads to acute coma and may result in prolonged disorder of consciousness (pDOC). We aimed to determine whether right median nerve electrical stimulation is a safe and effective treatment for accelerating emergence from coma after TBI. METHODS: This randomised controlled trial was performed in 22 centres in China. Participants with acute coma at 7-14 days after TBI were randomly assigned (1:1) to either routine therapy and right median nerve electrical stimulation (RMNS group) or routine treatment (control group). The RMNS group received 20 mA, 300 μs, 40 Hz stimulation pulses, lasting 20 s per minutes, 8 h per day, for 2 weeks. The primary outcome was the proportion of patients who regained consciousness 6 months post-injury. The secondary endpoints were Glasgow Coma Scale (GCS), Full Outline of Unresponsiveness scale (FOUR), Coma Recovery Scale-Revised (CRS-R), Disability Rating Scale (DRS) and Glasgow Outcome Scale Extended (GOSE) scores reported as medians on day 28, 3 months and 6 months after injury, and GCS and FOUR scores on day 1 and day 7 during stimulation. Primary analyses were based on the intention-to-treat set. RESULTS: Between March 26, 2016, and October 18, 2020, 329 participants were recruited, of whom 167 were randomised to the RMNS group and 162 to the control group. At 6 months post-injury, a higher proportion of patients in the RMNS group regained consciousness compared with the control group (72.5%, n = 121, 95% confidence interval (CI) 65.2-78.7% vs. 56.8%, n = 92, 95% CI 49.1-64.2%, p = 0.004). GOSE at 3 months and 6 months (5 [interquartile range (IQR) 3-7] vs. 4 [IQR 2-6], p = 0.002; 6 [IQR 3-7] vs. 4 [IQR 2-7], p = 0.0005) and FOUR at 28 days (15 [IQR 13-16] vs. 13 [interquartile range (IQR) 11-16], p = 0.002) were significantly increased in the RMNS group compared with the control group. Trajectory analysis showed that significantly more patients in the RMNS group had faster GCS, CRS-R and DRS improvement (p = 0.01, 0.004 and 0.04, respectively). Adverse events were similar in both groups. No serious adverse events were associated with the stimulation device. CONCLUSION: Right median nerve electrical stimulation is a possible effective treatment for patients with acute traumatic coma, that will require validation in a confirmatory trial