Counting Experiments Research Articles

How many syllables do you hear? Korean perception of English coda plosives

AbstractThis study investigates the direct perception of foreign structures in loanword phonology, concentrating on whether Korean listeners unnecessarily perceive an illusory vowel after a word-final plosive of English since a word-final voiceless plosive is permissible in their native language. In a syllable counting experiment, Korean participants listened to English nonce forms ending in a plosive and they indicated the number of syllables in each word. This study examined six different linguistic factors that may contribute to the perception of loanword borrowers including release, voicing and place of coda plosive, vowel tenseness, final stress and word size. The current results show that Korean listeners do perceive an extra syllable when the English pseudoword ends in a released plosive or in a syllable containing a tense vowel. This finding supports the hypothesis of adaptation in perception that unmotivated vowel insertion results from the misperception of English words rather than from a production grammar maintaining perceptual similarity between the English form and Korean pronunciation.

Theoretical analysis of argon 2p states' density ratios for nanosecond plasma optical emission spectroscopy

A theoretical analysis of excited argon state densities responsible for optical emission spectra of atmospheric pressure argon plasma is presented for its use in plasma diagnostics. Nanosecond pulsed barrier discharges are simulated using spatially one- and two-dimensional fluid-Poisson models using the reaction kinetics model presented by Stankov et al. [1], which considers all ten argon 2p states (Paschen notation) separately. The very first (single) discharge and repetitive discharges with frequencies from 5 kHz to 100 kHz are considered. A semi-automated procedure is utilized to find appropriate 2p states for electric field determination using an intensity ratio method, which is based on a time-dependent collisional-radiative model. The fluid simulations in combination with the semi-automated procedure are used to quantify the sensitivity of selected 2p-state ratios to given preionization of the gas. A highly sensitive time-correlated single photon counting experiment shows clearly that the selected ratio is sensitive to the electric field variation in the streamer head, yet additional calibration is needed for absolute values determination. Different approaches for effective lifetime determination are tested and applied also to measured data. The influence of radial and axial 2p state density integration on the intensity ratio method is discussed. The above mentioned models and procedures result in a flexible theory-based methodology applicable for development of new diagnostic techniques.

First-in-human study of 99mTc-labeled fucoidan, a SPECT tracer targeting P-selectin

BackgroundActivation of endothelial cells and platelets in atherothrombosis is characterized by upregulation of P-selectin. As a consequence, P-selectin represents a potential target for molecular imaging to identify thrombosis at an early stage. Fucoidan is a polysaccharide ligand extracted from brown algae with nanomolar affinity for P-selectin. This first-in-human study evaluated in healthy volunteers the safety, whole-body biodistribution, and dosimetry of 99mTc-fucoidan (Good Manufacturing Practices grade). We also investigated whether we could observe binding of 99mTc-fucoidan to human thrombi ex vivo and in vivo. In ten healthy volunteers, conjugate whole-body scans were performed up to 24 h following intravenous injection of 99mTc-fucoidan (370 MBq). Moreover, 99mTc-fucoidan uptake in ex vivo human thrombi (n = 11) was measured by gamma counting. Additionally, three patients with a newly diagnosed deep vein thrombosis (DVT) were subjected to 99mTc-fucoidan SPECT/CT imaging.Results99mTc-fucoidan was well tolerated in all participants without any drug-related adverse events. The total-body absorbed dose in males was comparable to females (0.012 ± 0.004 vs. 0.011 ± 0.005 mSv/MBq; p = 0.97). Gamma counting experiments demonstrated binding of tracer to ex vivo human thrombi that was 16% lower after blocking with a natural P-selectin ligand, Sialyl Lewis X. 99mTc-fucoidan demonstrated specific uptake at the thrombus site in one out of three scanned patients with DVT.Conclusions99mTc-Fucoidan has a favorable biodistribution and safety profile. 99mTc-fucoidan exhibited specific binding to human thrombi in both in vivo and ex vivo settings. Nonetheless, the in vivo results do not support further clinical investigation of 99mTc-fucoidan as an imaging modality for DVT. Other clinical implementations of a technetium− 99m-labeled P-selectin tracer should be considered.Trial registration: Clinicaltrials,NCT03422055.Registered 01/15/2018. URL: https://clinicaltrials.gov/study/NCT03422055Landelijk Trial Register, NL7739. Registered 4/2/2019 . https://onderzoekmetmensen.nl/en/trial/26785

NIM–TTL and TTL–NIM converters for single particle pulse counting experiments

Abstract Low-cost interface circuits are detailed that convert negative-going NIM signals to TTL and TTL to NIM, using high-speed digital and analogue components to convert between formats. By converting NIM pulses derived from single particle detectors to TTL logic, the large suite of TTL compatible devices available to manipulate, control and measure TTL signals can be exploited. Conversion of TTL pulses to NIM allows them to be used to trigger timing events, e.g. in time of flight measurements and coincidence studies. Examples using these techniques are presented.

Synthesis, Molecular Docking, and Anticancer Screening of Ester-Based Thiazole Derivatives.

This study investigates the potential of five compounds as novel anticancer agents. We examined their efficacy, mechanisms of action, and impact on various cancer cell lines, through a comprehensive set of experiments. Notably, compound 3e demonstrated superior activity compared to the positive control cisplatin, with a GI50 value of 6.3±0.7 μM against the breast cancer cell line (MCF-7). Compound 3b also displayed remarkable growth inhibition, yielding GI50 values of 8.7±0.2 μM (MCF-7) and 8.9±0.5 μM against the colon cancer cell line (HCT-116). Cell count experiments further confirmed the potent inhibitory effects of compounds 3e, 3b, and 3c on MCF-7 and HCT-116 cell growth. Compound 3e demonstrated a reduction of 55-60 % at GI50 and complete inhibition (100 %) at 2x GI50. Compound 3b exhibited 50-55 % reduction (GI50) and 90-95 % inhibition (2x GI50) in HCT-116 cells. Compound 3c displayed 75-80 % inhibition (2x GI50) and 35-40 % inhibition (GI50) in HCT-116 cells. In-depth mechanistic investigations unveiled valuable insights into the mode of action of compound 3e. The cell-cycle assay demonstrated G2/M phase arrest, DNA damage, and caspase-mediated apoptosis in both MCF-7 and HCT-116 cells. Caspase activation indicated a significant increase in apoptosis following exposure to compound 3e. Furthermore, compound 3e induced reactive oxygen species (ROS) production, influencing HCT-116 and MCF-7 cells differently. Elevated ROS production in HCT-116 cells and distinct effects in MCF-7 cells contribute to a deeper understanding of the cytotoxic mechanisms of compound 3e. Overall, these findings highlight the potential of the investigated compounds, particularly compound 3e, as effective inducers of apoptosis in cancer cells. Mechanistic insights into cell cycle arrest, caspase-mediated apoptosis, and ROS modulation provide a comprehensive understanding of their cytotoxic effects. This study offers significant contribution to the development of promising anticancer agents and their therapeutic applications.

Effects of 4-hydroxy-2(3H)-benzoxazolone on proliferation and apoptosis of pancreatic cancer L3.6 cells

This study explored the effects of 4-hydroxy-2(3H)-benzoxazolone(HBOA) on the proliferation and apoptosis of pancreatic cancer cells and its molecular mechanism. The L3.6 cells cultured in vitro were treated with HBOA of 0-1.0 mmol·L~(-1). The cell viability was detected by the cell counting kit-8(CCK-8) method, and the half inhibitory concentration(IC_(50)) was analyzed to determine the drug concentration and time. The cell morphology was observed under an inverted microscope and by acridine orange(AO) staining. The ability of proliferation and self-renewal were evaluated through live cell counting and colony formation experiments. The cell cycle progression and cell apoptosis rate were detected by flow cytometry. The morphology of cell apoptosis was observed by scanning electron microscopy. The mRNA expression of proliferating cell nuclear antigen(PCNA), cyclinA1, cyclinA2, cyclin dependent kinase 2(CDK2), and cyclin dependent kinase inhibitor 1A(P21) were determined by qPCR. The level of reactive oxygen species(ROS), lipid peroxide, and mitochondrial membrane potential were measured by flow cytometry. The activity of protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway was detected by Western blot. Compared with the control group, the cells treated with HBOA exhibited a significant decrease in viability. Then the optimal concentration and intervention time of HBOA were determined to be 0.4 mmol·L~(-1), 0.6 mmol·L~(-1), and 48 h. Compared with the control group, groups with HBOA of 0.4 mmol·L~(-1 )and 0.6 mmol·L~(-1) showed a significant suppression in cell proliferation and colony formation ability, down-regulated mRNA of PCNA, cyclinA1, cyclinA2, and CDK2, up-regulated P21 mRNA, S-phase cell cycle arrest, and increased cell apoptosis rate. There was an appearance of apoptotic bodies, increased ROS and lipid peroxide, decreased mitochondrial membrane potential(with a significant decrease in 0.6 mmol·L~(-1) group), and down-regulated p-Akt and p-mTOR proteins. The results show that HBOA inhibits the proliferation of pancreatic cancer L3.6 cells and induces cell apoptosis, which may be related to the increase in reactive oxygen species and the inhibition of the Akt/mTOR pathway.

MapReduce Algorithm on a Serverless Platform

Using the MapReduce (MR) programming technique, large-scale data processing tasks can be divided into tasks that are easier to manage and independent. The serverless implementation's performance is better than the non-serverless MR model when the model parameters are altered in the experiment. Optimizing the use of the MR model on the serverless platform can be achieved by taking into account the relationship between implementation efficiency and platform settings. Additionally, it can act as a source of inspiration for future serverless hardware support configurations. Also, the serverless platform demonstrates how it improves the effectiveness of resource utilization in machine learning training. The intended result can then be achieved by combining the results of these concurrently operating jobs on server clusters. This work adapts MR, a popular big data processing framework, to the serverless platform, emphasizing realization simulation principles and services, and then uses the results in a word count experiment. The experiment uses a word count of about eleven thousand words to evaluate how well MR is implemented on Alibaba Cloud. It is validated for execution time on the platform with varying Central Processing Unit (CPU) core counts, memory configurations, and worker counts. By testing several platform configurations, it is found that the memory configuration has very little impact on the model's execution time, while the size of the CPU core has a considerable impact on reaction time relative to the number of workers.

Research on Dynamic Pig Counting Method Based on Improved YOLOv7 Combined with DeepSORT.

A pig inventory is a crucial component of achieving precise and large-scale farming. In complex pigsty environments, due to pigs' stress reactions and frequent obstructions, it is challenging to count them accurately and automatically. This difficulty contrasts with most current deep learning studies, which rely on overhead views or static images for counting. This research proposes a video-based dynamic counting method, combining YOLOv7 with DeepSORT. By utilizing the YOLOv7 network structure and optimizing the second and third 3 × 3 convolution operations in the head network ELAN-W with PConv, the model reduces the computational demand and improves the inference speed without sacrificing accuracy. To ensure that the network acquires accurate position perception information at oblique angles and extracts rich semantic information, we introduce the coordinate attention (CA) mechanism before the three re-referentialization paths (REPConv) in the head network, enhancing robustness in complex scenarios. Experimental results show that, compared to the original model, the improved model increases the mAP by 3.24, 0.05, and 1.00 percentage points for oblique, overhead, and all pig counting datasets, respectively, while reducing the computational cost by 3.6 GFLOPS. The enhanced YOLOv7 outperforms YOLOv5, YOLOv4, YOLOv3, Faster RCNN, and SSD in target detection with mAP improvements of 2.07, 5.20, 2.16, 7.05, and 19.73 percentage points, respectively. In dynamic counting experiments, the improved YOLOv7 combined with DeepSORT was tested on videos with total pig counts of 144, 201, 285, and 295, yielding errors of -3, -3, -4, and -26, respectively, with an average accuracy of 96.58% and an FPS of 22. This demonstrates the model's capability of performing the real-time counting of pigs in various scenes, providing valuable data and references for automated pig counting research.

Deep learning-based recognition system for pashto handwritten text: benchmark on PHTI.

This article introduces a recognition system for handwritten text in the Pashto language, representing the first attempt to establish a baseline system using the Pashto Handwritten Text Imagebase (PHTI) dataset. Initially, the PHTI dataset underwent pre-processed to eliminate unwanted characters, subsequently, the dataset was divided into training 70%, validation 15%, and test sets 15%. The proposed recognition system is based on multi-dimensional long short-term memory (MD-LSTM) networks. A comprehensive empirical analysis was conducted to determine the optimal parameters for the proposed MD-LSTM architecture; Counter experiments were used to evaluate the performance of the proposed system comparing with the state-of-the-art models on the PHTI dataset. The novelty of our proposed model, compared to other state of the art models, lies in its hidden layer size (i.e., 10, 20, 80) and its Tanh layer size (i.e., 20, 40). The system achieves a Character Error Rate (CER) of 20.77% as a baseline on the test set. The top 20 confusions are reported to check the performance and limitations of the proposed model. The results highlight complications and future perspective of the Pashto language towards the digital transition.

Peptidoglycan synthesis drives a single population of septal cell wall synthases during division in Bacillus subtilis

Bacterial cell division requires septal peptidoglycan (sPG) synthesis by the divisome complex. Treadmilling of the essential tubulin homologue FtsZ has been implicated in septal constriction, though its precise role remains unclear. Here we used live-cell single-molecule imaging of the divisome transpeptidase PBP2B to investigate sPG synthesis dynamics in Bacillus subtilis. In contrast to previous models, we observed a single population of processively moving PBP2B molecules whose motion is driven by peptidoglycan synthesis and is not associated with FtsZ treadmilling. However, despite the asynchronous motions of PBP2B and FtsZ, a partial dependence of PBP2B processivity on FtsZ treadmilling was observed. Additionally, through single-molecule counting experiments we provide evidence that the divisome synthesis complex is multimeric. Our results support a model for B. subtilis division where a multimeric synthesis complex follows a single track dependent on sPG synthesis whose activity and dynamics are asynchronous with FtsZ treadmilling.

High expression of TBRG4 in relation to unfavorable outcome and cell ferroptosis in hepatocellular carcinoma

BackgroundHepatocellular carcinoma (HCC) is the most common type of malignant liver tumor with poor prognosis. In this study, we investigated the expression of transforming growth factor beta regulator 4 (TBRG4) in HCC and its effects on the proliferation, invasion, and metastasis of HCC cells, and analyzed the possible molecular mechanisms.MethodDownloading the expression and clinical information of HCC samples in the TCGA database, analyzing the expression differences of TBRG4 by bioinformatics methods, analyzing the clinical relevance and prognostic significance. Performing GO, KEGG and GSEA enrichment analysis on the TBRG4-related gene set in patient HCC tissues. Applying cell counting, scratch test and Transwell experiment to study the biological function of TBRG4 in HCC. Mitochondrial membrane potential, apoptosis and ROS levels were evaluated to assess cell iron death. Western blot, RT-PCR, laser confocal microscopy and co-immunoprecipitation were used to detect and analyze the downstream signaling pathways and interacting molecules of TBRG4.ResultsBioinformatics analysis revealed that TBRG4 was abnormally highly expressed in HCC tumor tissues and was associated with poor prognosis and metastasis in HCC patients. GO and KEGG functional enrichment analysis showed that TBRG4 was related to oxidative stress and NADH dehydrogenase (ubiquinone) activity. GSEA enrichment analysis showed that TBRG4 was associated with Beta catenin independent wnt signaling and B cell receptor. Functional experiments confirmed that knocking down TBRG4 could inhibit the proliferation, migration, and invasion of HCC cells. Mechanistically, TBRG4 inhibited the function of HCC cells through the DDX56/p-AKT/GSK3β signaling pathway. In addition, interference with TBRG4 expression could reduce the mitochondrial membrane potential and accumulate ROS in HCC cells, leading to increased ferroptosis. Co-IP analysis showed that TBRG4 specifically bound to Beclin1.ConclusionTBRG4 is highly expressed in HCC tumor tissues and is associated with poor prognosis. It may regulate the proliferation, invasion, and metastasis of HCC cells through the DDX56/p-AKT/GSK3β signaling pathway. TBRG4 may interact with Beclin1 to regulate the ferroptosis of HCC cells.

Abstract B072: Loss of the tumor and metastasis suppressor RasGAP DAB2IP mediates therapeutic resistance in HER2+ breast cancer

Abstract Resistance to HER2 inhibitors remains a clinical challenge in HER2+ breast cancer. Therefore, there is an urgent need to 1) understand the mechanisms that underlie resistance to these current treatments and 2) develop improved, and more importantly, curative combination therapies. We previously showed that the RasGAP DAB2IP is a tumor and metastasis suppressor in breast cancer. Interestingly, we have now generated robust data demonstrating that the loss of DAB2IP also mediates therapeutic resistance in HER2+ breast cancer. First, we genetically ablated DAB2IP in multiple HER2+ breast cancer cell lines and performed manual counting experiments after 6 days of HER2 TKI treatment. In all cell lines, DAB2IP knockdown conferred resistance to HER2 inhibitors. Since HER2 inhibitors are known to induce both apoptosis and senescence, we next investigated how DAB2IP loss specifically affects these phenotypes. Interestingly, DAB2IP knockdown prevented TKI-induced caspase-3/7 activation, measured by Incucyte live cell imaging, and enabled the regrowth of cells in long-term 10-day treatment and drug washout experiments, monitored by Incucyte or crystal violet staining. Mechanistically, stable knockdown of DAB2IP reduced the suppression of the AKT and ERK pathways and the induction of the pro-apoptotic protein Bim upon HER2i treatment. In addition, cell cycle progression pathways were enriched in DAB2IP-deficient cells on lapatinib treatment and immunoblots revealed higher residual levels of pRb and low levels of p27 in these cells. Interestingly, the levels of these proteins were restored and regrowth in low density assays was inhibited with co-treatment with an NF-kB inhibitor. We next evaluated the relevance of DAB2IP loss in mediating HER2 inhibitor resistance in a SUM190 orthotopic xenograft model. Importantly, while control tumors regressed upon lapatinib treatment, DAB2IP-deficient tumors did not regress and grew with kinetics comparable to vehicle-treated tumors after few days on treatment. Further understanding the pathways deregulated by DAB2IP loss upon HER2i treatment will be essential for developing effective therapeutic strategies to combat resistance induced by DAB2IP loss. Citation Format: Naiara Perurena, Amy Schade, Natalie Pilla, Patrick Loi, Carrie Rodriguez, Alycia Gardner, Karen Cichowski. Loss of the tumor and metastasis suppressor RasGAP DAB2IP mediates therapeutic resistance in HER2+ breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr B072.

Enhanced photocatalytic and antimicrobial activities of carbon-dots nanozymes modulated with P-doping

Bacterial contamination poses a serious and global threats to public health. Due to broad-band antimicrobial capacity and unspecific tissue interaction, antibacterial nanomaterials offer promising alternative and green strategies to resist bacterial contamination. Herein, an enhanced metal-free carbon-dots nanozymes, i.e., m-aminophenol based phosphoric acid doping carbon dots (P-CDs), with excellent photoresponse enzyme-like properties are designed through a simple one-pot hydrothermal method for highly effective antimicrobial. Developed P-CDs exhibited superior photoresponse oxidase-like activity, which may attribute to the higher crystallinity conferred by P-doping. Interestingly, the generation of superoxide radicals (O2•−) under photocatalytic for such P-CDs exhibited excellent antimicrobical effects with a high maximum reaction rate (33.3 × 10-8 M/s). Notably, P-CDs effectively inhibited the growth of both Gram-positive and Gram-negative bacteria under a 1 W white LED irradiation, especially against Gram-positive Staphylococcus aureus (S. aureus). Plate counting experiments confirmed that 75 μg/mL P-CDs could kill 98 % of E. coli when illuminated for 60 min, however, 50 μg/mL P-CDs could inhibit 100 % of S. aureus for the same illumination time. Surely, our metal-free carbon dots nanozyme provides a strong and viable alternative for the management of bacterial contamination in food security and environmental preservation, as well as a theoretical basis for P-doping to promote photocatalytic activities of carbon-dots.

From Cultured Chats to the Chirrups of Choo-Choo-Da-Choos, or How We Found a Key to the Gate of Eden

Reinvention of the form of expression is a conceptual approach characteristic for the evolution of all arts. This research study provides one such step forward in the advancement of scientific paper, a standard form of expression in natural sciences, toward more progressive terrains. The paper adopts the form of a theatrical play where a scientific family of four attempts to find the way around a writer’s block (Act I). Their idealess sense of confinement is overcome through arts or, more specifically, through scientific research implementing the aesthetics of music as its key element. The characters record compositions differing in the degrees of consonance, rhythmicity and tonality, and play them to birds to observe their vocal and behavioral responses (Act II). They note that the birdsong volume drops significantly when the piano tunes, especially the compositionally complex ones, are played to the birds and increases signifcantly when the white noise of ocean waves is played. The bird count experiment proves indirectly that the birdsong volume reduction is an auditory response of the birds to music rather than the effect of their migrating to or away from the sound source. This science with the flavor of art elevates the characters beyond the limits of their confinement and they, exalted and uplifted, gain ideas for creative research in their own disciplines of biomedical science (Act III). They relinquish the newly gained freedom in favor of embracing love for humanity that underlies medical research and return to their confinement, where they conduct, in the coda of the paper, a series of experiments that venture beyond musicology and ornithology and fall in the domains of tissue engineering and drug delivery, but also psychology (Act III). First, sparrow nests are used as templates for tissue engineering constructs, which facilitate the growth of human fibroblastic cells better than their artificial, gridded replicas. Next, the shell of an Easter egg, predominantly calcite in composition, is used with success as a drug delivery carrier. Lastly, the bird nest drawing experiment demonstrates a greater mental health of children than that of the adults, correlating with one of the overarching points of the paper, which is to present children as a model for grownups to follow rather than the other way around. Over the course of the play, the characters discuss the value of the various fusions between art and science, question the limits of human language and repeatedly go off-topic to celebrate the freeness of the creative thinking process. The play reports on real-life research methods and scientific results through a narrative, without diminishing their empirical rigor and analytical accuracy.

From Cultured Chats to the Chirrups of Choo-Choo-Da-Choos, or How We Found a Key to the Gate of Eden

Reinvention of the form of expression is a conceptual approach characteristic for the evolution of all arts. This research study provides one such step forward in the advancement of scientific paper, a standard form of expression in natural sciences, toward more progressive terrains. The paper adopts the form of a theatrical play where a scientific family of four attempts to find the way around a writer’s block (Act I). Their idealess sense of confinement is overcome through arts or, more specifically, through scientific research implementing the aesthetics of music as its key element. The characters record compositions differing in the degrees of consonance, rhythmicity and tonality, and play them to birds to observe their vocal and behavioral responses (Act II). They note that the birdsong volume drops significantly when the piano tunes, especially the compositionally complex ones, are played to the birds and increases signifcantly when the white noise of ocean waves is played. The bird count experiment proves indirectly that the birdsong volume reduction is an auditory response of the birds to music rather than the effect of their migrating to or away from the sound source. This science with the flavor of art elevates the characters beyond the limits of their confinement and they, exalted and uplifted, gain ideas for creative research in their own disciplines of biomedical science (Act III). They relinquish the newly gained freedom in favor of embracing love for humanity that underlies medical research and return to their confinement, where they conduct, in the coda of the paper, a series of experiments that venture beyond musicology and ornithology and fall in the domains of tissue engineering and drug delivery, but also psychology (Act III). First, sparrow nests are used as templates for tissue engineering constructs, which facilitate the growth of human fibroblastic cells better than their artificial, gridded replicas. Next, the shell of an Easter egg, predominantly calcite in composition, is used with success as a drug delivery carrier. Lastly, the bird nest drawing experiment demonstrates a greater mental health of children than that of the adults, correlating with one of the overarching points of the paper, which is to present children as a model for grownups to follow rather than the other way around. Over the course of the play, the characters discuss the value of the various fusions between art and science, question the limits of human language and repeatedly go off-topic to celebrate the freeness of the creative thinking process. The play reports on real-life research methods and scientific results through a narrative, without diminishing their empirical rigor and analytical accuracy.

Hsa_circ_0007460 affects the survival of intracellular Mycobacterium tuberculosis by regulating autophagy and apoptosis of macrophages.

Circular RNA (circRNA) is a category of non-coding RNAs characterized by the absence of a 5'-cap and 3'-poly(A) tail, and participates in the physiological processes of various human diseases. Nonetheless, the diagnostic and functional significance of circRNAs in active pulmonary tuberculosis (ATB) remains uncertain. Consequently, the purpose of this study is to investigate whether hsa_circ_0007460 can be employed as a potential diagnostic biomarker in ATB patients and explore its function. The result of real-time quantitative fluorescent PCR (RT-qPCR) validated a notable increase in the expression of hsa_circ_0007460 in the peripheral blood of 32 ATB patients, as well as in THP-1 human macrophages infected with Bacillus Calmette Guerin (BCG) which is an attenuated strain of Mycobacterium bovis. Additionally, the receiver operating curve (ROC) illustrated that the area under the ROC curve (AUC), sensitivity and specificity were 0.7474, 76.67%, and 78.13% respectively. RNase R, Actinomycin D and other experiments confirmed that hsa_circ_0007460 was stabler than its linear mRNA, indicating that hsa_circ_0007460 has potential as a diagnostic biomarker of ATB. Furthermore, Western blot (WB), Cell Counting Kit-8 (CCK-8), plate counting, and immunofluorescence experiments revealed that hsa_circ_0007460 could regulate apoptosis and autophagy of macrophages. The downstream miRNAs and mRNAs were subsequently predicted using bioinformatics, and the hsa circ 0007460/hsa-miR-3127-5p/PATZ1 axis was built. These above results suggest that hsa_circ_0007460 is substantially up-regulated in the peripheral blood of patients with ATB and can be utilized as a potential diagnostic biomarker. In addition, hsa_circ_0007460 can promote apoptosis of macrophages and inhibit autophagy of macrophages, thereby promoting the survival of BCG.

Preparation, characterization, and Staphylococcus aureus biofilm elimination effect of baicalein-loaded tyrosine/hyaluronic acid/β-cyclodextrin-grafted chitosan nano-delivery system

Staphylococcus aureus (S. aureus) is an important cause of infections associated with implanted medical devices due to the formation of bacterial biofilm, which can prevent the penetration of drugs, thus posing a serious multi-drug resistance. Methicillin-resistant Staphylococcus aureus (MRSA) is one of them. In order to enhance the biofilm elimination effect of Baicalein (BA), a BA-loaded Tyr/HA/CD-CS nano-delivery system was successfully prepared using β-cyclodextrin grafted with chitosan (CD-CS), Hyaluronic Acid (HA), and D-Tyrosine (D-Tyr). The Tyr/HA/CD-CS-BA-NPs have a uniform particle size distribution with a particle size of 238.1 ± 3.06 nm and a PDI of 0.130 ± 0.02. The NPs showed an obvious inhibitory effect on planktonic bacteria with a MIC of 12.5 μg/mL. In vivo and in vitro tests showed that the NPs could enhance the elimination effect of BA on the MRSA biofilm. The results of Confocal Laser Scanning Microscopy (CLSM), Live & Dead Kit, and colony count experiments illustrated that Tyr/HA/CD-CS-BA-NPs could enhance the permeability of drugs to the biofilm and improve the ability to kill the biofilm bacteria, which may be an important mechanism to enhance the elimination of the MRSA biofilm. These findings will help develop new, effective medicaments for treating bacterial biofilm infections.

Managing photoinduced electronic interactions on a molybdenum disulfide/diblock copolymer/anionic porphyrin nanoensemble

Exploring the modular nature of MoS2 and its ability to be easily functionalized, both in covalent and non-covalent ways and with a variety of chemical species, a MoS2/diblock copolymer system was conceptualized. As the constituents of this diblock copolymer, the poly(benzyl methacrylate), abbreviated as pBzMA, block was designed to serve as the immobilization point of the polymeric chains onto the MoS2 lattice, taking advantage of the S-π non-covalent interactions between phenyl rings and ample S atoms, while the quaternized poly(dimethylamino-ethyl methacrylate), abbreviated as QpDMAEMA, was incorporated as the second block to capture the tetrasulfo-tetraphenyl anionic porphyrin, abbreviated as TPPS4 via electrostatic interactions with the ammonium groups of the quaternized polymeric chain. The so-formed MoS2/QpDMAEMA-b-pBzMA/TPPS4 ensemble was characterized and assessed via spectroscopic (UV-Vis), electron microscopy imaging (TEM) and dynamic light scattering (DLS) techniques. Markedly, efficient quenching of the characteristic fluorescence of the porphyrin species by the MoS2 nanosheets was observed, suggesting the development of photoinduced charge-transfer phenomena within the MoS2/QpDMAEMA-b-pBzMA/TPPS4 ensemble. This was further corroborated by time-correlated single photon counting (TCSPC) and blank experiments, signifying the role of Coulombic interactions for bringing into electronic communication the porphyrin moieties with MoS2. Overall, this rationally designed nanoensemble based on non-covalently modified MoS2 with polymers and photoactive species sets the foundations for inexpensive yet potent systems to be used in energy-related applications.

Time Intervals Produced by Silent Chronometric Counting are Involuntarily Affected by Number Word Magnitudes

Abstract Chronometric counting is a method to approximate the duration of a time interval by keeping track of the accumulation of its one-second subintervals. The ordinality of the number words is instrumental to this method, but whether also the magnitudes of these words affect the approximations remains unclear. The participants performed self-initiated and silent chronometric counting in different directions to produce target intervals prospectively. Two experiments were conducted. In Experiment 1, counting from 1- to 6-s target intervals started or stopped at zero. In Experiment 2, 1- or 2-s target intervals were counted with low-magnitude (1–3) or high-magnitude (4–6) number words. The participants were found to overproduce target intervals towards their shorter durations (Experiments 1 and 2) and, at a trend level, with downward rather than upward counting (Experiment 1 but not Experiment 2). They also produced target intervals as longer in duration with high- than low-magnitude number words (Experiment 2). The main findings suggest an involuntary magnitude effect of endogenously activated number words on subjective time.

Class I HDAC inhibition reduces DNA damage repair capacity of MYC-amplified medulloblastoma cells

PurposeMYC-driven Group 3 medulloblastoma (MB) (subtype II) is a highly aggressive childhood brain tumor. Sensitivity of MYC-driven MB to class I histone deacetylase inhibitors (HDACi) has been previously demonstrated in vitro and in vivo. In this study we characterize the transcriptional effects of class I HDACi in MYC-driven MB and explore beneficial drug combinations.MethodsMYC-amplified Group 3 MB cells (HD-MB03) were treated with class I HDACi entinostat. Changes in the gene expression profile were quantified on a microarray. Bioinformatic assessment led to the identification of pathways affected by entinostat treatment. Five drugs interfering with these pathways (olaparib, idasanutlin, ribociclib, selinexor, vinblastine) were tested for synergy with entinostat in WST-8 metabolic activity assays in a 5 × 5 combination matrix design. Synergy was validated in cell count and flow cytometry experiments. The effect of entinostat and olaparib on DNA damage was evaluated by γH2A.X quantification in immunoblotting, fluorescence microscopy and flow cytometry.ResultsEntinostat treatment changed the expression of genes involved in 22 pathways, including downregulation of DNA damage response. The PARP1 inhibitors olaparib and pamiparib showed synergy with entinostat selectively in MYC-amplified MB cells, leading to increased cell death, decreased viability and increased formation of double strand breaks, as well as increased sensitivity to additional induction of DNA damage by doxorubicin. Non-MYC-amplified MB cells and normal human fibroblasts were not susceptible to this triple treatment.ConclusionOur study identifies the combination of entinostat with olaparib as a new potential therapeutic approach for MYC-driven Group 3 MB.