Naringenin Against Cadmium Toxicity in Fibroblast Cells: An Integrated Network Pharmacology and In Vitro Metabolomics Approach.

Cadmium, a heavy metal, disrupts cellular homeostasis and is highly toxic, with no effective treatments currently available against its toxicity. According to studies, phytochemicals provide a promising strategy for mitigating cadmium toxicity. Naringenin (NG), a potent antioxidant found primarily in citrus fruits, showed protective properties against cadmium toxicity in rats. Nonetheless, the precise mechanism of cadmium cytotoxicity in fibroblasts remains unknown. This study evaluated NG against cadmium (CdCl2) toxicity utilizing network pharmacology and in silico molecular docking, and was further validated experimentally in rat fibroblast F111 cells. Using network pharmacology, 25 possible targets, including the top 10 targets of NG against cadmium, were identified. Molecular docking of interleukin 6 (IL6), the top potential target with NG, showed robust binding with an inhibition constant (Ki) of 58.76 µM, supporting its potential therapeutic potential. Pathway enrichment analysis suggested that "response to reactive oxygen species" and "negative regulation of small molecules metabolic process" were the topmost pathways targeted by NG against cadmium. In vitro analysis showed that NG (10 µM) attenuated CdCl2-induced oxidative stress by reducing altered intracellular ROS, mitochondrial mass, and membrane potential. Also, NG reversed CdCl2-mediated nuclear damage, G2/M phase arrest, and apoptosis. GC/MS-based metabolomics of F111 cells revealed CdCl2 reduced cholesterol levels, which led to alterations in primary bile acid, steroid and steroid hormone biosynthesis pathways, whereas, NG restored these alterations. In summary, combined in silico and in vitro analysis suggested that NG protected cells from CdCl2 toxicity by mitigating oxidative stress and metabolic pathway alterations, providing a comprehensive understanding of its protective mechanisms against cadmium-induced toxicity.

Effect of Lipid Corona on Phenylalanine-Functionalized Gold Nanoparticles to Develop Stable and Corona-Free Systems.

Conventional gold nanoparticles (Au NPs) have many limitations, such as aggregation and subsequent precipitation in the medium of high ionic strength and protein molecules. Furthermore, when exposed to biological fluids, nanoparticles form a protein corona, which controls different biological processes such as the circulation lifetime, drug release profile, biodistribution, and in vivo cellular distribution. These limitations reduce the functionality of Au NPs in targeted delivery, bioimaging, gene delivery, drug delivery, and other biomedical applications. To circumvent these problems, there are numerous attempts to design corona-free and stable nanoparticles. Here, we report for the first time that lipid corona (coating of lipid) formation on phenylalanine-functionalized Au NPs (AuPhe NPs) imparts excellent stability against the high ionic strength of bivalent metal ions, amino acids, and proteins of different charges as compared to bare nanoparticles. Moreover, this work is focused on the ability of lipid corona formation on AuPhe NPs to prevent protein adsorption in the presence of cell culture medium (CCM), oppositely charged protein (e.g., histone 3), and human serum albumin (HSA). The results demonstrate that the lipid corona successfully protects the AuPhe NPs from protein adsorption, leading to the development of corona-free character. This unique achievement has profound implications for enhancing the biomedical utility and safety of these nanoparticles.

Machine Learning Classification of Body Part, Imaging Axis, and Intravenous Contrast Enhancement on CT Imaging.

Purpose: The development and evaluation of machine learning models that automatically identify the body part(s) imaged, axis of imaging, and the presence of intravenous contrast material of a CT series of images. Methods: This retrospective study included 6955 series from 1198 studies (501 female, 697 males, mean age 56.5 years) obtained between January 2010 and September 2021. Each series was annotated by a trained board-certified radiologist with labels consisting of 16 body parts, 3 imaging axes, and whether an intravenous contrast agent was used. The studies were randomly assigned to the training, validation and testing sets with a proportion of 70%, 20% and 10%, respectively, to develop a 3D deep neural network for each classification task. External validation was conducted with a total of 35,272 series from 7 publicly available datasets. The classification accuracy for each series was independently assessed for each task to evaluate model performance. Results: The accuracies for identifying the body parts, imaging axes, and the presence of intravenous contrast were 96.0% (95% CI: 94.6%, 97.2%), 99.2% (95% CI: 98.5%, 99.7%), and 97.5% (95% CI: 96.4%, 98.5%) respectively. The generalizability of the models was demonstrated through external validation with accuracies of 89.7 - 97.8%, 98.6 - 100%, and 87.8 - 98.6% for the same tasks. Conclusions: The developed models demonstrated high performance on both internal and external testing in identifying key aspects of a CT series.

Biogenic carbon dots: a novel mechanistic approach to combat multidrug-resistant critical pathogens on the global priority list.

This study delves into investigating alternative methodologies for anti-microbial therapy by focusing on the mechanistic assessment of carbon dots (CDs) synthesized from F. benghalensis L. extracts. These biogenic CDs have shown remarkable broad-spectrum anti-bacterial activity even against multi-drug resistant (MDR) bacterial strains, prompting a deeper examination of their potential as novel anti-microbial agents. The study highlights the significant detrimental impact of CDs on bacterial cells through oxidative damage, which disrupts the delicate balance of ROS control within the cells. Notably, even at low doses, the anti-bacterial activity of CDs against MDR strains of P. aeruginosa and E. cloacae is highly effective, demonstrating their promise as potent antimicrobial agents. The research sheds light on the capacity of CDs to generate ROS, leading to membrane lipid peroxidation, loss of membrane potential, and rupture of bacterial cell membranes, resulting in cytoplasmic leakage. SEM and TEM analysis revealed time-dependent cell surface, morphological, and ultrastructural changes such as elongation of the cells, irregular surface protrusion, cell wall and cell membrane disintegration, internalization, and aggregations of CDs. These mechanisms offer a comprehensive explanation of how CDs exert their anti-bacterial effects. We also determined the status of plasma membrane integrity and evaluated live (viable) and dead cells upon CD exposure by flow cytometry. Furthermore, comet assay, biochemical assays, and SDS PAGE identify DNA damage, carbohydrate and protein leakage, and distinct differences in protein expression, adding another layer of understanding to the mechanisms behind CDs' anti-bacterial activity. These findings pave the way for future research on managing ROS levels and developing CDs with enhanced anti-bacterial properties, presenting a breakthrough in anti-microbial therapy.

Prevalence and predictors of shared decision-making in goals-of-care clinician-family meetings for critically ill neurologic patients: a multi-center mixed-methods study.

BACKGROUND: Shared decision-making is a joint process where patients, or their surrogates, and clinicians make health choices based on evidence and preferences. We aimed to determine the extent and predictors of shared decision-making for goals-of-care discussions for critically ill neurological patients, which is crucial for patient-goal-concordant care but currently unknown. METHODS: We analyzed 72 audio-recorded routine clinician-family meetings during which goals-of-care were discussed from seven US hospitals. These occurred for 67 patients with 72 surrogates and 29 clinicians; one hospital provided 49/72 (68%) of the recordings. Using a previously validated 10-element shared decision-making instrument, we quantified the extent of shared decision-making in each meeting. We measured clinicians' and surrogates' characteristics and prognostic estimates for the patient's hospital survival and 6-month independent function using post-meeting questionnaires. We calculated clinician-family prognostic discordance, defined as ≥ 20% absolute difference between the clinician's and surrogate's estimates. We applied mixed-effects regression to identify independent associations with greater shared decision-making. RESULTS: The median shared decision-making score was 7 (IQR 5-8). Only 6% of meetings contained all 10 shared decision-making elements. The most common elements were "discussing uncertainty"(89%) and "assessing family understanding"(86%); least frequent elements were "assessing the need for input from others"(36%) and "eliciting the context of the decision"(33%). Clinician-family prognostic discordance was present in 60% for hospital survival and 45% for 6-month independent function. Univariate analyses indicated associations between greater shared decision-making and younger clinician age, fewer years in practice, specialty (medical-surgical critical care > internal medicine > neurocritical care > other > trauma surgery), and higher clinician-family prognostic discordance for hospital survival. After adjustment, only higher clinician-family prognostic discordance for hospital survival remained independently associated with greater shared decision-making (p = 0.029). CONCLUSION: Fewer than 1 in 10 goals-of-care clinician-family meetings for critically ill neurological patients contained all shared decision-making elements. Our findings highlight gaps in shared decision-making. Interventions promoting shared decision-making for high-stakes decisions in these patients may increase patient-value congruent care; future studies should also examine whether they will affect decision quality and surrogates' health outcomes.

Effect of differential deprivation of nutrients on cellular proliferation, oxidative stress, mitochondrial function, and cell migration in MDA-MB-231, HepG2, and HeLa cells.

Cancerous cells display metabolic engineering through enhanced utilization of nutrients to support their increased requirements for proliferation, bioenergetics, biosynthesis, redox homeostasis, and cell signaling. To investigate the extent to which malignant cells rely on glycolysis and glutaminolysis, the effects of differential deprivation of nutrients such as d-glucose, l-glutamine, and pyruvate on proliferation, morphology, cell cycle, oxidative stress, mitochondrial function, autophagic vacuole formation, and migration in MDA-MB-231, HepG2, and HeLa cells were investigated in this study. Cell viability assay,  cell morphology, and ATP assay showed higher dependence of MDA-MB-231 and HepG2 cells on glucose and glutamine, respectively, for cell survival, growth, ATP production, and proliferation, while HeLa cells were equally dependent on both. However, the combination of all three nutrients displayed maximum proliferation. Differential deprivation of glucose in the absence of glutamine resulted in G0/G1 plus G2/M arrest in MDA-MB-231, whereas G0/G1 arrest in HepG2 and S-phase arrest in HeLa cells occurred at 48 h. Although the differential withdrawal of nutrients revealed a varying degree of effect dependent on cell type, nutrient type, nutrient concentrations, and deprivation time, a general trend of increased oxidative stress, loss of mitochondrial membrane potential, and ATP and antioxidant (GSH) depletion led to mitochondrial dysfunction in all three cell lines and inhibition of cell migration in MDA-MB-231 and HeLa cells at 48 h. Extreme deprivation of nutrients formed autophagic vacuoles. Importantly, normal cells (HEK293) remained unaffected under most of the nutrient-deprived conditions examined. This study enhances our understanding of the impact of differential nutrient deprivation on critical characteristics of cancer cells, contributing to the development of metabolism-based effective anticancer strategies. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03759-w.

Nanobiotechnological approaches in anticoagulant therapy: The role of bioengineered silver and gold nanomaterials.

Nanotechnology is a novel area that has exhibited various remarkable applications, mostly in medicine and industry, due to the unique properties coming with the nanoscale size. One of the notable medical uses of nanomaterials (NMs) that attracted enormous attention recently is their significant anticoagulant activity, preventing or reducing coagulation of blood, decreasing the risk of strokes, heart attacks, and other serious conditions. Despite successful in vitro experiments, in vivo analyses are yet to be confirmed and further research is required to fully prove the safety and efficacy of nanoparticles (NPs) and to introduce them as valid alternatives to conventional ineffective anticoagulants with various shortcomings and side-effects. NMs can be synthesized through two main routes, i.e., the bottom-up route as a more preferable method, and the top-down route. In numerous studies, biological fabrication of NPs, especially metal NPs, is highly suggested given its eco-friendly approach, in which different resources can be employed such as plants, fungi, bacteria, and algae. This review discusses the green synthesis and characterization of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) as two of the most useful metal NPs, and also their alloys in different studies focussing on their anticoagulant potential. Challenges and alternative approaches to the use of these NPs as anticoagulants have also been highlighted.

Phytochemical Profiling of Tupistra nutans Wall. ex Lindl. Inflorescence Extract and Evaluation of Its Antioxidant Activity and Toxicity in Hepatocarcinoma (HepG2) and Fibroblast (F111) Cells.

Tupistra nutans Wall. ex Lindl. is a medicinal plant found in the Eastern Himalayan region. Besides being used as a folk medicine for pain and high blood sugar, its inflorescence is consumed as a vegetable. However, its medicinal properties have not been proven in vitro and in vivo till now. Therefore, in this study, we reported the phytochemicals present in the methanolic extract of Tupistra nutans Wall. ex Lindl. inflorescence (METNI) and its comparative effect in liver carcinoma HepG2 cells against non-cancerous murine fibroblast F111 cells. Phytochemical profiling by gas chromatography-mass spectrometry (GC-MS) analysis showed that METNI was rich in unsaturated fatty acids, vitamin E, and anticancer compounds like diosgenin, linoleic acid, and palmitoleic acid. METNI was found to have in vitro antioxidant property as determined by DPPH and pyrogallol methods, and UV protection property as investigated by fluorescence-based and spectrophotometric methods. MTT assay revealed METNI caused significantly more cell proliferation inhibition in HepG2 (IC50 = 138 µg/ml) compared to F111 (IC50 = 347 µg/ml) cells. Although in both HepG2 and F111 cells METNI showed significant antioxidant activity, it led to intracellular ROS generation and cell cycle alteration at higher exposure. The obtained results suggest that Tupistra nutans can be a promising application for anticancer drug and skin care product development, but can be harmful if overconsumed.

Investigation of temperature sensitivity of a MEMS gravimeter based on geometric anti-spring.

This paper describes a technique for temperature sensitivity or thermal sag measurements of a geometric anti-spring based microelectromechanical system (MEMS) gravimeter (Wee-g). The Wee-g MEMS gravimeter is currently fabricated on a (100) silicon wafer using standard micro-nano fabrication techniques. The thermal behavior of silicon indicates that the Young's modulus of silicon decreases with increase in temperature (∼64 ppm/K). This leads to a softening of the silicon material, resulting in the proof mass displacing (or sagging) under the influence of increasing temperature. It results in a change in the measured gravity, which is expressed as temperature sensitivity in terms of change in gravity per degree temperature. The temperature sensitivity for the silicon based MEMS gravimeter is found to be 60.14-64.87, 61.76, and 62.76 µGal/mK for experimental, finite element analysis (FEA) simulation, and analytical calculations, respectively. It suggests that the gravimeter's temperature sensitivity is dependent on the material properties used to fabricate the MEMS devices. In this paper, the experimental measurements of thermal sag are presented along with analytical calculations and simulations of the effect using FEA. The bespoke optical measurement system to quantify the thermal sag is also described. The results presented are an essential step toward the development of temperature insensitive MEMS gravimeters.

Evaluation of antimicrobial, anticancer potential and Flippase induced leakage in model membrane of Centella asiatica fabricated MgONPs.

The use of chemically synthesized nanoparticles and crude plant extracts as antimicrobial -anticancer agents have many limitations. In this study, we have used Centella asiatica extract (CaE) having relatively less explored but tremendous medicinal properties, as reducing and stabilizing agents to green synthesize magnesium oxide nanoparticles (MgONPs) using magnesium nitrate. In comparison to the bulk material, capabilities of Ca-MgONPs as an improved antibacterial, antifungal, and anticancer agent in human prostatic carcinoma cells (PC3), as well as membranolytic capability in model cell membrane, were studied. The phyto-functionalized Ca-MgONPs were characterized using UV-Visible spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Energy Dispersive X-Ray Spectroscopy (EDX), X-ray Diffraction (XRD), Fourier Transform Infra-Red Spectroscopy (FT-IR) and Atomic Force Microscopy (AFM). Observation of characteristic peaks by spectroscopic and microscopic analysis confirmed the synthesis of Ca-MgONPs. The Ca-MgONPs showed broad spectrum of bactericidal activity against both gram-positive and gram-negative bacteria and fungicidal activity against two species of the Candida fungus. The Ca-MgONPs also exhibited dose-dependent and selective inhibition of proliferating PC3 cells with IC50 of 123.65 ± 4.82 µg/mL at 24 h, however, without having any cytotoxicity toward non-cancerous HEK293 cells. Further studies aimed at understanding the probable mechanism of toxicity of Ca-MgONPs in PC3 cells, the results indicated a significant reduction in cell migration capacities, increment in cytosolic ROS, loss of mitochondrial transmembrane potential, DNA damage and S-phase cell cycle arrest. Ca-MgONPs also induced pore formation in a synthetic large unilamellar vesicle. Thus, Ca-MgONPs might be useful in the effective management of several human pathogens of concern and some more cancer types.

Low-dose exposure to phytosynthesized gold nanoparticles combined with glutamine deprivation enhances cell death in the cancer cell line HeLa via oxidative stress-mediated mitochondrial dysfunction and G0/G1 cell cycle arrest.

Cancer cells use nutrients like D-glucose (Glc) and L-glutamine (Q) more efficiently for their development. This increased nutritional dependency of malignant cells has been commonly employed in various in vitro and in vivo models of anticancer therapies. This study utilized a combination of a low dose (25 µg mL-1) of S2, a phytosynthesized gold nanoparticle (AuNP) that was previously proven to be non-toxic, and deprivation of extracellular glutamine as an anticancer strategy in the human cervical cancer cell line HeLa. We discovered that 24 h Q deprivation led to a less significant decrease in the viability of HeLa cells while a low dose of S2 caused a non-significant reduction in the viability of HeLa cells. However, combining these two treatments resulted in highly significant inhibition of cell growth, as measured by the MTT test and morphological examination. Glutamine starvation in HeLa cells was found to induce cellular uptake of S2 via clathrin-mediated endocytosis, thus facilitating the improved antitumor effects of the combined treatment. Flow cytometry-based assays using fluorescent probes H2DCFDA and MitoSOX Red confirmed that this combination therapy involved the development of oxidative stress conditions owing to a surplus of cytosolic reactive oxygen species (cytoROS) and mitochondrial superoxide (mtSOX) generation. Furthermore, the investigated combinatorial treatment also indicated mitochondrial inactivity and disintegration, as evidenced by the drop in the mitochondrial membrane potential (Δψm) and the decrease in the mitochondrial mass (mtMass) in a flow-cytometric assay utilizing the probes. Tetramethylrhodamine ethyl ester and MitoTracker Green FM, respectively. Cell cycle arrest in the G0/G1 phase, induction of cell death via apoptosis/necrosis, and inhibition of migration capacities of HeLa cells were also seen after the combined treatment. Thus, this research provides insight into a new combinatorial approach for reducing the dose of nanoparticles and increasing their efficacy to better inhibit the growth of human cervical cancer cells by leveraging their extracellular glutamine dependence.

A 19 day earth tide measurement with a MEMS gravimeter.

The measurement of tiny variations in local gravity enables the observation of subterranean features. Gravimeters have historically been extremely expensive instruments, but usable gravity measurements have recently been conducted using MEMS (microelectromechanical systems) sensors. Such sensors are cheap to produce, since they rely on the same fabrication techniques used to produce mobile phone accelerometers. A significant challenge in the development of MEMS gravimeters is maintaining stability over long time periods, which is essential for long term monitoring applications. A standard way to demonstrate gravimeter stability and sensitivity is to measure the periodic elastic distortion of the Earth due to tidal forces-the Earth tides. Here, a 19 day measurement of the Earth tides, with a correlation coefficient to the theoretical signal of 0.975, has been presented. This result demonstrates that this MEMS gravimeter is capable of conducting long-term time-lapse gravimetry, a functionality essential for applications such as volcanology.

Characterization of mechanical properties of materials using ultrasound broadband spectroscopy.

This article explores the characterization of homogenous materials (metals, alloys, glass and polymers) by a simple broadband ultrasonic interrogation method. The novelty lies in the use of ultrasound in a continuous way with very low input power (0 dBm or less) and analysis of the transmitted acoustic wave spectrum for material property characterization like speed of sound, density and dimensions of a material. Measurements were conducted on various thicknesses of samples immersed in liquid where continuous-wave, frequency swept ultrasonic energy was incident normal to the sample surface. The electro-acoustic transmission response is analyzed in the frequency domain with respect to a specifically constructed multi-layered analytical model. From the acoustic signature of the sample materials, material properties such as speed of sound and acoustic impedance can be calculated with experimentally derived values found to be in general agreement with the literature and with pulse-echo technique establishing the basis for a non-contact and non-destructive technique for material characterization. Further, by looking at the frequency spacing of the peaks of water when the sample is immersed, the thickness of the sample can be calculated independently from the acoustic response. This technique can prove to be an effective non-contact, non-destructive and fast material characterization technique for a wide variety of materials.

Investigating biomechanical noise in neuroblastoma cells using the quartz crystal microbalance.

Quantifying cellular behaviour by motility and morphology changes is increasingly important in formulating an understanding of fundamental physiological phenomena and cellular mechanisms of disease. However, cells are complex biological units, which often respond to external environmental factors by manifesting subtle responses that may be difficult to interpret using conventional biophysical measurements. This paper describes the adaptation of the quartz crystal microbalance (QCM) to monitor neuroblastoma cells undergoing environmental stress wherein the frequency stability of the device can be correlated to changes in cellular state. By employing time domain analysis of the resulting frequency fluctuations, it is possible to study the variations in cellular motility and distinguish between different cell states induced by applied external heat stress. The changes in the frequency fluctuation data are correlated to phenotypical physical response recorded using optical microscopy under identical conditions of environmental stress. This technique, by probing the associated biomechanical noise, paves the way for its use in monitoring cell activity, and intrinsic motility and morphology changes, as well as the modulation resulting from the action of drugs, toxins and environmental stress.