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Abstract: In this study, the electric cell-substrate impedance sensing (ECIS) system was used to study the cellular activities of oral squamous cell carcinoma (OSCC) cells in a real-time and label-free manner. Various cellular activities, including cell adhesion, spreading, proliferation, and drug-induced apoptosis and inhibition of apoptosis, were monitored. A linear relationship was found between the impedance-based cell index and the cell number in the range of 3500 to 35,000 cells/well. Anti-cancer drug-cisplatin-induced OSCC cell apoptosis at the minimal concentration of 5μM after 20h of treatment and followed a linear dose-dependent manner in the concentration range from 10μM to 25μM. The inhibition of cisplatin-induced apoptosis by the carcinogen, nicotine, at concentrations from 0.1μM to 10μM was monitored. The most significant inhibitory effect of nicotine on cisplatin-induced apoptosis was observed at concentrations of 0.5–1μM. The results obtained with impedance method correlated well with microscopic imaging analysis of cellular morphology and cell viability analysis. This study demonstrated that the impedance-based method can provide real-time information about the cellular activity of viable cells and detect drug-induced cellular activities much earlier than commonly used cell-based image analysis. This impedance-based method has the potential to provide a useful analytical approach for cancer research. [Copyright &y& Elsevier]

Abstract: The performance of a low conductive growth medium (LCGM) (conductivity of <1300 μS) was evaluated for its ability to support growth of food borne bacterial pathogens including Listeria monocytogenes and to determine the expression of the two key virulence proteins in L. monocytogenes for possible applications in an impedance-based microfluidic biochip detection platform. Growth of Listeria was monitored spectrophotometrically and the lag phase, generation time, growth rate and maximum population density were determined using the Gompertz equation. LCGM had a lag phase of 2.3 h and showed a higher cell density compared to Luria Bertini (LB) broth. Length of lag phase was highly dependent on initial inoculum concentrations. The changes in conductivity with respect to growth in the low conductive medium were monitored using a conductivity probe. L. monocytogenes growth could be detected within 2 h (0.1 mS) in LCGM and within 6 h in LB. The performance of the media was also evaluated for the recovery of Listeria cells exposed to various stresses as 42 °C for 1, 2 or 6 h, an osmotic stress in 10.5% NaCl, an acidic stress at pH 2, 3 or 5 and a combined stress of 10.5% NaCl, pH 5 and 1 h exposure at 42 °C. The recovery rate was comparable with that of Tryptic soy broth containing yeast extract (TSBYE). L. monocytogenes in LCGM supported the expression of two key virulence markers, actin polymerization protein (ActA) and internalin B (InlB), which could be detected using specific antibodies. In general LCGM also supported the growth of several other bacterial species suggesting its implication in microbial quality monitoring of products. In conclusion, LCGM is a sensitive low conductive medium that supports the growth as well as the expression of virulence markers for potential applications in sensitive detection of L. monocytogenes or other food borne pathogens in impedance-based sensor platform. [Copyright &y& Elsevier]

Carbon dots (CDots) are strongly absorptive over the visible spectrum, with the effective photon-harvesting driving rich excited state processes and properties. In this work, spectroscopic probing of these processes and properties is coupled with the evaluation of the photoinduced bactericidal function of CDots, aimed toward making correlations among the findings from the former and those from the latter on the inactivation of bacterial pathogens. Within the general mechanistic framework for CDots, the observed effective and efficient antibacterial activities of the CDots under visible light are attributed to major contributions by the initially photo-generated electrons and holes that are trapped at passivated surface defect sites of the dots, in addition to the traditional reactive oxygen species (ROS) produced in the emissive excited states from the recombination of the redox pairs. Such major contributions to the inactivation of the bacteria by the separated redox species in CDots can not be quenched by ROS scavengers commonly used in the study of photodynamic effects with classical molecular photosensitizers, thus making light-activated CDots uniquely potent antimicrobial agents. The characteristic features of photoexcited CDots and their related mechanistic implications are discussed in reference to the similar behaviors and mechanistic model of conventional semiconductor quantum dots. Image 1 [ABSTRACT FROM AUTHOR]

[Display omitted] • Carbon dots (CDots) exploit unique properties of small carbon nanoparticles; • CDots with N -ethylcarbazole (NEC) functionalization are very stable; • NEC-CDots are excellently compatible with poly(N -vinylcarbazole) (PVK); • Optical properties of NEC-CDots in PVK matrix are no different from those in solution; • PVK/NEC-CDots nanocomposites are valuable to electronic/optoelectronic applications. The adducts of N -ethylcarbazole (NEC) to small carbon nanoparticles prepared by microwave-assisted thermal processing are carbon dots (CDots) with NEC for the dot surface functionalization, thus NEC-CDots, which were shown previously for their high stability in the dot structure and properties. In this study, the optical absorptions and photoexcited state properties of NEC-CDots in solution and in poly(N -vinylcarbazole) (PVK) film matrix were evaluated by using optical spectroscopy methods, including those for fluorescence decays and lifetimes. The results suggest that the properties are largely unchanged from solution phase to the polymer matrix environment, very positive to their expected optoelectronic applications. [ABSTRACT FROM AUTHOR]

Background: Cell-based analysis may have the potential for using patient-derived tissues/cells as a basis for a more direct prediction of therapeutic outcomes.Methods: This study focused on the application of the Real Time Cell Electronic Sensing (RT-CES) system for screening effective anticancer drugs to different types of prostate cancers. The goal was to demonstrate the potential application of such technology toward the suitability in selection of personalized medicine. Three prostate cancer cell lines, DU145, LNCaP, and PC-3, and a normal prostate cell line were used. Docetaxel, Carboplatin, Abiraterone Acetate, Mitoxantrone, Sunitinib Malate were used as testing drugs.Results: Cellular adhesion, proliferation and drug induced cellular responses were monitored by the RTCES system, and the results were correlated to the well-established cell viability tests. Identification of an effective drug from a panel of available anticancer drugs to a specific cancer cell line, or testing the effectiveness of a certain drug to a panel of prostate cancer cell lines was demonstrated. Cellular resistance to a drug at single dose and multiple dose challenges was monitored by RT-CES measurement, while such resistance was not detectable by endpoint cell viability assays.Conclusions: The results of this study highlighted the advantages of such real-time impedance-based sensing system for applications in chemotherapy reagent selection. [ABSTRACT FROM AUTHOR]

A simple yet consequential modification was made to the popular carbonization processing of citric acid - polyethylenimine precursor mixtures to produce carbon dots (CDots). The modification was primarily on pushing the carbonization processing a little harder at a higher temperature, such as the hydrothermal processing condition of around 330 °C for 6 h. The CDots thus produced are comparable in spectroscopic and other properties to those obtained in other more controlled syntheses including the deliberate chemical functionalization of pre-processed and selected small carbon nanoparticles, demonstrating the consistency in CDots and reaffirming their general definition as carbon nanoparticles with surface passivation by organic or other species. Equally significant is the finding that the modified processing of citric acid - polyethylenimine precursor mixtures could yield CDots of record-setting fluorescence performance, approaching the upper limit of being quantitatively fluorescent. Thus, the reported work serves as a demonstration on not only the need in selecting the right processing conditions and its associated opportunities in one-pot syntheses of CDots, but also the feasibility in pursuing the preparation of quantitatively fluorescent CDots, which represents an important milestone in the development and understanding of these fluorescent carbon nanomaterials. [ABSTRACT FROM AUTHOR]

Background: After parents raised concerns about potential lead (Pb) contamination of calcium carbonate for treatment of hyperphosphatemia in chronic kidney disease (CKD), we measured blood Pb using high-resolution sector field inductively coupled mass spectrometry in a quality-assurance investigation of ten pediatric dialysis patients (nine on hemodialysis) and six patients before dialysis. Methods: We assessed the kidney function as cystatin C estimated glomerular filtration rate (eGFR), blood Pb levels, calcium carbonate dose, and standard laboratory parameters, as well as Pb levels in the dialysis feed water. Results: Mean blood Pb concentration in the 16 pediatric CKD patients was 21.1 ± 15.8 µg/l with a maximum of 58 µg/l, which was significantly higher than that of 467 apparently healthy controls (median 6.35 µg/l, interquartile range 4.47, 8.71) and comparable to that of ten adult peritoneal dialysis (PD) patients. Lead levels correlated with red blood cell distribution width, eGFR, and calcium carbonate dose. Pb in dialysate feed water was always <0.00018 mg/l, which is below the accepted limit for water for dialysis of 0.005 mg/l. Conclusions: We found a high prevalence of elevated Pb levels in pediatric CKD patients that correlated with the calcium carbonate dose and GFR. Lead levels should be monitored in these patients. [ABSTRACT FROM AUTHOR]

• Carbon dots (CDots) exploit unique properties of small carbon nanoparticles; • Carbon/semiconductor hybrid dots substantially expand the CDots platform; • Carbon/TiO 2 dots in different compositions/configurations can be designed and prepared; • The hybrid dots can be explored for different photoexcited state properties; • The hybrid dots can serve as effective photodynamic antibacterial agents. Carbon/TiO 2 hybrid dots with the general configuration of nanoscale carbon domains incorporated with TiO 2 nanoparticles of different sizes (8 and 25 nm in diameter) were prepared and characterized, and their optical and photodynamic properties were studied. The properties were found to be affected dramatically by the relative sizes and configurations of the nanoscale carbon domains and TiO 2 nanoparticles in the hybrid dots, as reflected by the observed major difference in their photodynamic inactivation of bacteria under visible light. Mechanistic implications of the results are discussed. [ABSTRACT FROM AUTHOR]

Introduction/aims: Line blot (LB) is in widespread use for myositis antibody detection. Yet, studies of its positive predictive value (PPV) in patients with suspected idiopathic inflammatory myopathy (IIM), which would be of particular relevance to neuromuscular clinicians, are lacking. We aimed to determine the PPV of myositis antibody LB testing in patients with suspected IIM, and examine whether PPV was significantly impacted by intensity of antibody positivity., Methods: This was a retrospective study of patients who underwent myositis antibody LB testing for suspected IIM between March 2019 and August 2022., Results: Of 70 patients who underwent testing for suspected IIM and had positive myositis antibody LB results, 43 (61%) were female and the median age was 61 years (range: 10-83 years). Forty-four were classified as true-positives, yielding a PPV of 63%. The PPV of patients with weak-positive myositis antibody results (14/30, 47%) was significantly lower than the PPV of patients with moderate-positive or strong-positive myositis antibody results (30/40, 75%) (p = .02)., Discussion: Our study found that myositis antibody LB testing in patients with suspected IIM had a modest PPV, underscoring the need for antibody interpretation in the context of all available clinical and ancillary test data to avoid misdiagnosis. The significantly lower PPV in patients with weak-positive results emphasizes the particular importance of clinical correlation in such patients. Further study into the diagnostic performance of various LBs for myositis antibody detection is needed to inform their interpretation in clinical practice., (© 2024 The Authors. Muscle & Nerve published by Wiley Periodicals LLC.)

Introduction: In January 2023, our laboratory began performing serum myelin oligodendrocyte glycoprotein antibody (anti-MOG) titers by fixed cell-based assay (CBA). As a quality assurance (QA) assessment, we evaluated titer positive predictive value (PPV) as well as impact of sample collection timing on titers., Methods: Among patients who underwent antibody titers to distinguish between low-positive (<1:100) and clear-positive (≥1:100) anti-MOG, records were reviewed to classify results as true-positive (TP) or false-positive (FP) and facilitate PPV calculation. Timing of sample collection relative to administration of immunotherapy and symptom onset was determined for TP results., Results: Overall PPV of anti-MOG was 70/85 (82%). The PPV of low-positive anti-MOG was significantly lower than clear-positive anti-MOG (72% vs. 95%, p  = 0.009). The difference in PPV between low-positive and clear-positive anti-MOG was significant among adults tested, but not children. Among patients with TP anti-MOG, the proportion who received immunotherapy prior to sample collection was significantly higher and median time from symptom onset to sample collection was significantly longer for low-positive compared to clear-positive results., Conclusion: Overall PPV of anti-MOG testing by fixed CBA was reasonably high. The PPV of low-positive anti-MOG was significantly lower than clear-positive anti-MOG. This was driven by the significantly lower PPV of low-positive anti-MOG in adults, possibly reflecting the lower prevalence of MOG antibody-associated disease among adults tested. Timing of sample collection relative to administration of immunotherapy and symptom onset may substantially impact titers, indicating that testing should ideally be performed prior to immunotherapy and close to time of attack., Competing Interests: DLR has received research support from MS Canada, the National MS Society, Consortium of MS Centers, and Roche Canada. She has received speaking or consulting fees from Amgen, Alexion, Biogen, EMD Serono, Novartis, Roche, Sanofi Aventis, and Touch IME. EAY has received research funding from the National MS Society, Consortium of MS Centers, CIHR, NIH, OIRM, SCN, CBMH Chase an Idea, SickKids Foundation, Rare Diseases Foundation, MS Scientific Foundation, McLaughlin Centre, Leong Center, Peterson Foundation; Investigator initiated research funding from Biogen; Scientific advisory: Hoffman-LaRoche; Speaker honoraria: Biogen, JHU, Saudi Epilepsy Society, NYU, MS-ATL, ACRS, PRIME, CNPS. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Budhram, Rotstein, Yang and Yeh.)

Background and Objectives: Ganglioside antibodies can help diagnose distinct acute and chronic inflammatory neuropathies including axonal variants of Guillain-Barre syndrome, Miller-Fisher syndrome (MFS), multifocal motor neuropathy, and chronic sensory ataxic neuropathies. Because ganglioside antibody testing may be routinely ordered in patients with suspected inflammatory neuropathy, we sought to evaluate its yield and utilization in clinical practice., Methods: We performed a retrospective chart review of all patients at London Health Sciences Centre who underwent ganglioside antibody testing between April 2019 and August 2023. The disease phenotype was determined for each patient, and the proportion of all tests that yielded a true-positive result was calculated. Ganglioside antibody positivity was classified as a true-positive result if the disease phenotype was robustly associated with the detected ganglioside antibody and there was no other more likely diagnosis., Results: We identified 92 patients who underwent ganglioside antibody testing. One patient (1%) was classified as having a true-positive result; this patient had GQ1b-IgG positivity with MFS. Among 92 patients tested, 20 patients (22%) had a disease phenotype that was considered to be robustly associated with ganglioside antibody positivity., Conclusions: The yield of ganglioside antibody testing in clinical practice is low. We found that this testing is frequently ordered in patients with disease phenotypes that are not robustly associated with ganglioside antibody positivity, indicating that suboptimal test utilization is a primary contributor to its low yield. Restricting ganglioside antibody testing to patients with characteristic disease phenotypes would be valuable to improving yield and utilization of this testing., Competing Interests: Declaration of competing interest Dr. Cathy Meng Fei Li has no disclosures to report. Dr. Yiu-Chia Chang has no disclosures to report. Dr. Liju Yang has no disclosures to report. Dr. Adrian Budhram reports that he holds the London Health Sciences Centre and London Health Sciences Foundation Chair in Neural Antibody Testing for Neuro-Inflammatory Diseases, and receives support from the Opportunities Fund of the Academic Health Sciences Centre Alternative Funding Plan of the Academic Medical Organization of Southwestern Ontario (AMOSO)., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Carbon-semiconductor hybrid quantum dots are classical carbon dots with core carbon nanoparticles doped with a selected nanoscale semiconductor. Specifically, on those with the nanoscale TiO 2 doping, denoted as C TiO2 -Dots, their synthesis and thorough characterization were reported previously. In this work, the C TiO2 -Dots were evaluated for their visible light-activated antibacterial function, with the results showing the effective killing of not only Gram-positive but also the generally more resistant Gram-negative bacteria. The hybrid dots are clearly more potent antibacterial agents than their neat carbon dot counterparts. Mechanistically, the higher antibacterial performance of the C TiO2 -Dots is attributed to their superior photoexcited state properties, which are reflected by the observed much brighter fluorescence emissions. Also considered and discussed is the possibility of additional contributions to the antibacterial activities due to the photosensitization of the nanoscale TiO 2 by its doped core carbon nanoparticles.

Extracellular adenosine triphosphate (ATP) is one of the most abundant biochemical constitutes within the stem cell microenvironment and is postulated to play critical roles in cell migration. However, it is unclear whether ATP regulates the cell migration of CD34 + vascular wall-resident stem/progenitor cells (VW-SCs) and participates in angiogenesis. Therefore, the biological mechanisms of cell migration mediated by ATP was determined by in vivo subcutaneous matrigel plug assay, ex vivo aortic ring assay, in vitro transwell migration assay, and other molecular methods. In the present study, ATP dose-dependently promoted CD34 + VW-SCs migration, which was more obviously attenuated by inhibiting or knocking down P2Y2 than P2Y6. Furthermore, it was confirmed that ATP potently promoted the migration of resident CD34 + cells from cultured aortic artery rings and differentiation into endothelial cells in matrigel plugs by using inducible lineage tracing Cd34 -CreER T2 ; R26-tdTomato mice, whereas P2Y2 and P2Y6 blocker greatly inhibited the effect of ATP. In addition, ATP enhanced the protein expression of stromal interaction molecule 1 (STIM1) on cell membrane, blocking the calcium release-activated calcium (CRAC) channel with shSTIM1 or BTP2 apparently inhibited ATP-evoked intracellular Ca 2+ elevation and channel opening, thereby suppressing ATP-driven cell migration. Moreover, extracellular signal-regulated protein kinase (ERK) inhibitor PD98059 and p38 inhibitor SB203580 remarkably inhibited ERK and p38 phosphorylation, cytoskeleton rearrangement, and subsequent cell migration. Unexpectedly, it was found that knocking down STIM1 greatly inhibited ATP-triggered ERK/p38 activation. Taken together, it was suggested that P2Y2 signaled through the CRAC channel mediated Ca 2+ influx and ERK/p38 pathway to reorganize the cytoskeleton and promoted the migration of CD34 + VW-SCs. NEW & NOTEWORTHY In this study, we observed that the purinergic receptor P2Y2 is critical in the regulation of vascular wall-resident CD34 + cells' migration. ATP could activate STIM1-mediated extracellular Ca 2+ entry by triggering STIM1 translocation to the plasma membrane, and knockdown of STIM1 prevented ERK/p38 activation-mediated cytoskeleton rearrangement and cell migration.

High anti-GAD65 levels associate with core manifestations of GAD65 neurological autoimmunity. ELISA cut-offs for high anti-GAD65 levels (>10,000 IU/ml in serum, >100 IU/ml in CSF) have been proposed that merit further evaluation. We reviewed patients who underwent anti-GAD65 ELISA for suspected autoimmune encephalitis and found values above these cut-offs to have a positive predictive value (PPV) for neurological autoimmunity of 88%. Anti-GAD65 values above proposed ELISA cut-offs have a reasonably high PPV for neurological autoimmunity in patients with suspected autoimmune encephalitis. Consideration of alternative diagnoses and corroboration with CSF can help flag potentially clinically irrelevant results and avoid patient misdiagnosis.

Carbon dots are defined as small carbon nanoparticles with effective surface passivation via organic functionalization. The definition is literally a description of what carbon dots are originally found for the functionalized carbon nanoparticles displaying bright and colorful fluorescence emissions, mirroring those from similarly functionalized defects in carbon nanotubes. In literature more popular than classical carbon dots are the diverse variety of dot samples from "one-pot" carbonization of organic precursors. On the two different kinds of samples from the different synthetic approaches, namely, the classical carbon dots versus those from the carbonization method, highlighted in this article are their shared properties and apparent divergences, including also explorations of the relevant sample structural and mechanistic origins for the shared properties and divergences. Echoing the growing evidence and concerns in the carbon dots research community on the major presence of organic molecular dyes/chromophores in carbonization produced dot samples, demonstrated and discussed in this article are some representative cases of dominating spectroscopic interferences due to the organic dye contamination that have led to unfound claims and erroneous conclusions. Mitigation strategies to address the contamination issues, including especially the use of more vigorous processing conditions in the carbonization synthesis, are proposed and justified., (© 2023 The Authors. Small published by Wiley-VCH GmbH.)

Background: Assessment of trace and toxic element status is important for the diagnosis and monitoring of several pediatric conditions. Elemental deficiency and toxicity have serious implications, particularly in pediatrics wherein risk is higher. Pediatric reference intervals (RIs) for trace elements and normal exposure limits for toxic elements are lacking on modern analytical systems. Herein, reference values were established for 13 plasma and 22 whole blood trace elements in the Canadian Laboratory Initiative on Pediatric Reference Intervals (CALIPER) cohort of healthy children and adolescents., Methods: Approximately 320 healthy children and adolescents were recruited with informed consent. Trace elements were measured in whole blood and plasma samples using 2 technologies: (a) triple quadrupole inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) (n = 172) and (b) high-resolution sector field ICPMS (HR-SF-ICPMS) (n =161). RIs and normal exposure limits were then established according to Clinical and Laboratory Standards Institute guidelines., Results: Of all elements assessed, none required sex partitioning and 8 required age partitioning (e.g., copper, manganese, and cadmium). Reference value distributions determined via ICP-MS/MS and HR-SF-ICPMS demonstrated excellent concordance, with few exceptions (e.g., molybdenum, cobalt, and nickel)., Conclusions: These data represent the first study wherein pediatric RIs and normal exposure limits were derived simultaneously on 2 different clinically validated MS platforms which provide urgently needed data to inform clinical decision-making for trace elements in pediatrics. Study findings suggest some trace elements require age-specific consideration for appropriate interpretation. Highly concordant observations across the 2 analytical methods also demonstrate the comparability and reliability of results obtained on both platforms., (© American Association for Clinical Chemistry 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: Cervical cancer (CC) is one of the main causes of death among gynecological malignancies. Patients with CC with lymph node metastasis (LNM) have poor prognoses. We investigated the risk factors and prognosis of LNM in patients with CC patients using data from the SEER database., Methods: We collected the information of cervical cancer patients registered in SEER database from 2010 to 2015. The dataset was divided into a training set and a validation set at a 7:3 ratio. LASSO regression analysis was used to evaluate risk factors for LNM in patients with CC. Using the results, we established a nomogram prediction model. C-index, ROC curves, calibration curves, decision curve analysis, and clinical impact curves were used to evaluate the prediction performance of the model., Results: We included 14,356 patients with CC in the analysis. Among these, 3997 patients were diagnosed with LNM. A training set (10,050 cases) and a validation set (4306 cases) were used for the following analysis. We established nomogram LNM prediction models for the patients with T 1-2 -stage CC. The C-indices for the internal and external validations of the prediction models were 0.758 and 0.744, respectively. In addition, we established a prognostic nomogram for all CC patients with LNM, and the internal and external validation C-indices were 0.763 and 0.737., Conclusion: We constructed a quantitative and visual predictive nomogram that predicted prognosis of patients with LNM in CC to provide clinicians with a reference for diagnosis and treatment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Ma, Yang and Liang.)

The prevention of viral transmission is an important step to address the spread of viral infections. Using the enveloped vesicular stomatitis virus (VSV) as a model, this study explored the antiviral functions of the specifically designed and prepared carbon dots (CDots). The CDots were prepared using small carbon nanoparticles with surface functionalization-passivation by oligomeric polyethylenimine (PEI). The results indicated that the PEI-CDots were readily activated by visible light to effectively and efficiently inactivate VSVs under various combinations of experimental conditions (viral titer, dot concentration, and treatment time). The photodynamically induced viral structural protein degradation and genomic RNA degradation were observed, suggesting the mechanistic origins, leading to the inactivation of virus. The results suggested CDots as a class of promising broad-spectrum antiviral agents for disinfection of viruses.

Carbon "quantum" dots or carbon dots (CDots) exploit and enhance the intrinsic photoexcited state properties and processes of small carbon nanoparticles via effective nanoparticle surface passivation by chemical functionalization with organic species. The optical properties and photoinduced redox characteristics of CDots are competitive to those of established conventional semiconductor quantum dots and also fullerenes and other carbon nanomaterials. Highlighted here are major advances in the exploration of CDots for their serving as high-performance yet nontoxic fluorescence probes for one- and multi-photon bioimaging in vitro and in vivo , and for their uniquely potent antimicrobial function to inactivate effectively and efficiently some of the toughest bacterial pathogens and viruses under visible/natural or ambient light conditions. Opportunities and challenges in the further development of the CDots platform and related technologies are discussed.

This study aimed to address the significant problems of bacterial biofilms found in medical fields and many industries. It explores the potential of classic photoactive carbon dots (CDots), with 2,2'-(ethylenedioxy)bis (ethylamine) (EDA) for dot surface functionalization (thus, EDA-CDots) for their inhibitory effect on B. subtilis biofilm formation and the inactivation of B. subtilis cells within established biofilm. The EDA-CDots were synthesized by chemical functionalization of selected small carbon nanoparticles with EDA molecules in amidation reactions. The inhibitory efficacy of CDots with visible light against biofilm formation was dependent significantly on the time point when CDots were added; the earlier the CDots were added, the better the inhibitory effect on the biofilm formation. The evaluation of antibacterial action of light-activated EDA-CDots against planktonic B. subtilis cells versus the cells in biofilm indicate that CDots are highly effective for inactivating planktonic cells but barely inactivate cells in established biofilms. However, when coupling with chelating agents (e.g., EDTA) to target the biofilm architecture by breaking or weakening the EPS protection, much enhanced photoinactivation of biofilm-associated cells by CDots was achieved. The study demonstrates the potential of CDots to prevent the initiation of biofilm formation and to inhibit biofilm growth at an early stage. Strategic combination treatment could enhance the effectiveness of photoinactivation by CDots to biofilm-associated cells., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Dong, Overton, Tang, Darby, Sun and Yang.)

Foodborne pathogens have long been recognized as major challenges for the food industry and repeatedly implicated in food product recalls and outbreaks of foodborne diseases. This study demonstrated the application of a recently discovered class of visible-light-activated carbon-based nanoparticles, namely, carbon dots (CDots), for photodynamic inactivation of foodborne pathogens. The results demonstrated that CDots were highly effective in the photoinactivation of Listeria monocytogenes in suspensions and on stainless steel surfaces. However, it was much less effective for Salmonella cells, but treatments with higher CDot concentrations and longer times were still able to inactivate Salmonella cells. The mechanistic implications of the observed different antibacterial effects on the two types of cells were assessed, and the associated generation of intracellular reactive oxygen species (ROS), the resulting lipid peroxidation, and the leakage of nucleic acid and proteins from the treated cells were analyzed, with the results collectively suggesting CDots as a class of promising photodynamic inactivation agents for foodborne pathogens. IMPORTANCE Foodborne infectious diseases have long been recognized as major challenges in public health. Contaminations of food processing facilities and equipment with foodborne pathogens occur often. There is a critical need for new tools/approaches to control the pathogens and prevent such contaminations in food processing facilities and other settings. This study reports a newly established antimicrobial nanomaterials platform, CDots coupled with visible/natural light, for effective and efficient inactivation of representative foodborne bacterial pathogens. The study will contribute to promoting the practical application of CDots as a new class of promising nanomaterial-based photodynamic inactivation agents for foodborne pathogens.

Neural antibodies have emerged as useful biomarkers in suspected autoimmune encephalitis. We reviewed results of neural antibody testing (anti-N-methyl D-aspartate receptor (NMDAR), leucine-rich glioma-inactivated protein (LGI1), contactin-associated protein-like 2 (CASPR2), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR), γ-aminobutyric acid type B receptor (GABA(B)R), dipeptidyl-peptidase-like protein-6 (DPPX), IgLON family member 5 (IgLON5) and glutamic acid decarboxylase-65 (GAD65)) using cell-based assays (CBAs) and tissue indirect immunofluorescence (TIIF) at our centre. Our findings suggest increased clinical sensitivity of CBA compared to TIIF. However, this may come at some expense to clinical specificity, as evidenced by possible false-positive results when weak serum positivity by CBA was observed for certain antibodies (i.e. anti-NMDAR, CASPR2). In such cases, correlation with serum TIIF, as well as CSF CBA and TIIF, aids in identifying true-positive results.

Protein electrophoresis is commonly used as an aid in the diagnosis of monoclonal gammopathies and is performed in many laboratories in Canada and throughout the world. However, unlike many other diagnostic tests, there is limited guidance for standardization and neither guidance nor specific recommendations for clinical reporting of serum (SPE) or urine (UPE) protein electrophoresis and immunotyping available in the literature. Therefore, a Canadian effort was undertaken to recommend standards that cover all aspects of clinical reporting with an ultimate goal towards reporting standardization. The Canadian Society of Clinical Chemists (CSCC) Monoclonal Gammopathy Interest Group (MGIG), which is composed of CSCC members with an interest in protein electrophoresis, has formed a Monoclonal Gammopathy Working Group (MGWG) to take initial steps towards standardization of SPE, UPE and immunotyping. Candidate standardization recommendations were developed, discussed and voted upon by the MGWG. Candidate recommendations that achieved 90% agreement are presented as consensus recommendations. Recommendations that did not achieve 90% consensus remain candidate recommendations and are presented with accompanying MGWG discussion. Eleven consensus recommendations along with candidate recommendations for nomenclature, protein fraction reporting, test utilization, interference handling and interpretive reporting options are presented., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Aims: Many of the secondary effects of high levels of molybdenum (Mo) overlap with symptoms commonly seen in pediatric patients with chronic kidney disease (CKD). We measured plasma Mo levels and examined the relationship between Mo levels and kidney function., Materials and Methods: The study was carried out at the London Health Sciences Centre in London, Ontario, Canada with 36 children and adolescents 4 - 18 years of age with CKD. There were 1 - 6 trace element measurements (Mo and copper (Cu)) per patient. We studied the proportion of patients with abnormal trace element levels and the relationship between trace element levels and estimated glomerular filtration rate (eGFR), calculated using the Filler formula. Plasma Mo and Cu levels were measured using High Resolution Sector Field Inductively Coupled Mass Spectrometry. Anthropomorphic data and blood parameters were collected from our electronic chart program., Results: Median eGFR was 51 mL/min/1.73m 2 (35, 75). Median Mo level was 2.00 µg/L (1.40, 2.88). 20 patients had at least one set of Mo levels above the published reference interval in either unit, and the results of 46% of the tests were above the interval. There was a strong negative correlation between the Mo levels and the eGFR (Spearman's r = -0.627, p < 0.0001)., Conclusions: Our study suggests that pediatric patients with CKD have elevated plasma levels of Mo, which may cause secondary effects commonly associated with CKD. The elevated Mo levels in our center's catchment area may cause an accumulation of this trace element in patients with impaired renal function.
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Multi-walled carbon nanotube (MWCNT) sheet was fabricated from a drawable MWCNT forest and then deposited on poly(methyl methacrylate) film. The film was further coated with a natural antimicrobial peptide nisin. We studied the effects of nisin coating on the attachment of Bacillus anthracis spores, the germination of attached spores, and the subsequent biofilm formation from attached spores. It was found that the strong adsorptivity and the super hydrophobicity of MWCNTs provided an ideal platform for nisin coating. Nisin coating on MWCNT sheets decreased surface hydrophobicity, reduced spore attachment, and reduced the germination of attached spores by 3.5 fold, and further inhibited the subsequent biofilm formation by 94.6% compared to that on uncoated MWCNT sheet. Nisin also changed the morphology of vegetative cells in the formed biofilm. The results of this study demonstrated that the anti-adhesion and antimicrobial effect of nisin in combination with the physical properties of carbon nanotubes had the potential in producing effective anti-biofilm formation surfaces., (Copyright © 2014. Published by Elsevier B.V.)

In this study, electric/electrochemical impedance spectroscopy and cyclic voltammetry were used to study the cellular activities of oral cancer cell line CAL 27, including the kinetics of cell adhesion, spreading, and cell proliferation on interdigitated microelectrodes (IMEs). Impedance spectra of CAL 27 cells on IMEs electrodes were obtained in cell growth medium and in 0.1 M PBS with 50 mM [Fe(CN)₆]³⁻/⁴⁻ as redox probe. Equivalent circuits were used to model both cases. In cell growth medium, impedance spectra allowed us to analyze the changes in capacitance and resistance due to cell attachment on the IMEs over the entire experiment period. It was found that cell spreading caused the most significant decrease in capacitance component and slight increase in resistance component. Impedance change at given frequencies (between 10 kHz to 100 kHz) was found to be linearly increased with increasing cell number of CAL 27 on the IMEs. In comparison with non-cancer oral epithelial cells (Het-1A), at equal cell number, cancer cells always generated impedance several folds higher than that of non-cancer cells. In the presence of [Fe(CN)₆]³⁻/⁴⁻, impedance spectra allowed us to analyze the change in electron transfer resistance of IMEs due to cell attachment, in which an increase trend was observed at 24 h with increasing cell number from 2500 cells to 10,000 cells on IMEs. Double layer capacitance was also affected by cell attachment, and a decrease in double layer capacitance was observed with increasing cell number on the electrodes. Cyclic voltammetric measurements correlated well with the impedance results. The results of this study demonstrated the use of electrochemical approaches to obtain and understand cellular behaviors/activities of oral cancer cells, potentially providing useful tools for cancer cell research.

Microlithographically fabricated interdigitated microsensor electrodes (IMEs) were cleaned, surface activated, chemically functionalized (amine) and derivatized with an Acrloyl-PEG-NHS to receive a spun-applied monomer cocktail of UV polymerizable monomer. IMEs were 2050.5, 1550.5, 1050.5 and 0550.5 possessing lines and spaces that were 20, 15, 10, and 5 μm respectively; 5 mm line lengths and were 50 lines on each opposing bus. Bioactive hydrogels were synthesized from spun-applied and UV-crosslinked tetraethyleneglycol diacrylate (TEGDA) (crosslinker), 2-hydroxyethylmethacrylate (HEMA), polyethyleneglycol(200) monomethacrylate (PEGMA), N-[tris(hydroxymethyl)methyl]-acrylamide (HMMA) and poly(HEMA) (MW 60,000) (viscosity modifier) and 2,2-dimethoxy-2-phenylacetophenone (DMPA) (photoinitiator) to produce a 5 μm thick p(HEMA-co-PEGMA-co-HMMA) hydrogel membrane on the IMEs. Unmodified and hydrogel coated IMEs where characterized by AC electrical impedance spectroscopy using 50 mV p-t-p over the frequency range from 10 Hz to 100 kHz in aqueous PBS 7.4 buffer and in buffer containing 50 mM [Fe(CN)(6)](3-/4- ) solution at RT. Impedimetric responses were found to scale with the device geometric parameters. Equivalent circuit modeling revealed deviations from ideality at lower device dimensions suggesting an implication of the substrate surface charge on the double layer capacitance of the electrodes. Diffusion coefficients derived from the Warburg component are in accord with literature values.

In this study, electrical impedance-based measurements were used to distinguish oral cancer cells and non-cancer oral epithelial cells based on their cellular activities on the microelectrodes in a real-time and label-free manner. CAL 27 and Het-1A cell lines were used as the models of oral cancer cells and non-cancer oral epithelial cells, respectively. Various cellular activities, including cell adhesion, spreading, and proliferation were monitored. We found that both the kinetics of cell spreading and the static impedance-based cell index were feasible to distinguish the two cell types. At each given cell number, CAL 27 cell spreading produced a smaller cell index change rate that was 60-70% of those of Het-1A cells. When cells were fully spread, CAL 27 cells generated a cell index more than four times greater than that of Het-1A cells. Since cell spreading and attachment occurs in the first few hours when they were cultured on the microelectrodes, this impedance-based method could be a rapid label-free and non-invasive approach to distinguish oral cancer cells from non-cancer oral epithelial cells. Cell viability analysis was performed along with the impedance-based analysis. Confocal microscopic imaging analysis showed the difference in cell morphology and the thickness of cell monolayers between the two cell types.

This study integrated dielectrophoresis (DEP) with non-flow through biochips to enhance the immuno-capture and detection of foodborne pathogenic bacteria. It demonstrated two major functions provided by DEP to improve the chip performance: (i) concentrating bacterial cells from the suspension to different locations on the chip surface by positive and negative DEP; (ii) making the cells in close contact with the immobilized antibodies on the chip surface so that immuno-capture efficiency can be dramatically enhanced. The microchip achieved the immuno-capture efficiencies of approximately 56.0% and approximately 64.0% to Salmonella cells with 15 and 30 min DEP, respectively, which were considerably higher than those of approximately 10.4% and approximately 17.6% for 15 and 30 min immuno-capture without DEP. The immuno-captured bacterial cells were detected by the sandwich format ELISA on the chips. The final absorbance signals were enhanced by DEP assisted immuno-capture by 64.7-105.2% for the samples containing 10(3)-10(6) cells/20 microl. The integration of DEP with the biochips has the potential to advance the chip-based immunoassay methods for microbial detection.

In this study, we present a new, simple and rapid impedance method to detect bacterial cells by making use of the impedance properties of bacterial cell suspensions using interdigitated microelectrodes. It was found that bacterial cell suspensions in deionized (DI) water with different cell concentrations could generate different electrical impedance spectral responses, whereas cell suspensions in phosphate buffered saline (PBS) solution could not produce any significant differences in impedance spectra in response to different cell concentrations. In DI water suspensions, impedance at 1 kHz decreased with the increasing cell concentrations in the suspensions. The impedance of cell suspensions in DI water was discussed and found that it was resulted from the cell wall charges and the release of ions or other osmolytes from the cells. A linear relationship between the impedance and the logarithmic value of the cell concentration was found in the cell concentration range from 10(6) to 10(10)cfu/ml, which can be expressed by a regression equation of Z(k Omega)=-2.06 logC (cells/20 microl)+5.23 with R(2)=0.98. The detection limit was calculated to be 3.45 x 10(6)cfu/ml, which is comparable with many label-free immunosensors for detection of pathogenic bacteria reported in the literature. To achieve the selectivity of this method, we also demonstrated the feasibility of integrating magnetic separation to this impedance method. This study has demonstrated that bacterial cell concentration can be inferred by measuring the impedance of cell suspensions in DI water. This new detection mechanism could be an alternative to current impedance methods that have been reported for the detection of bacterial cells, e.g. impedance microbiology and electrical/electrochemical impedance biosensors.

A quartz crystal microbalance (QCM) DNA sensor, based on the nanoparticle amplification method, was developed for detection of Escherichia coli O157:H7. A thiolated single-stranded DNA (ssDNA) probe specific to E. coli O157:H7 eaeA gene was immobilized onto the QCM sensor surface through self-assembly. The hybridization was induced by exposing the ssDNA probe to the complementary target DNA, and resulted in the mass change and therefore frequency change of the QCM. Streptavidin conjugated Fe(3)O(4) nanoparticles (average diameter=145 nm) were used as "mass enhancers" to amplify the frequency change. Synthesized biotinylated oligonucleotides as well as E. coli O157:H7 eaeA gene fragments (151 bases) amplified using asymmetric PCR with biotin labeled primers were tested. As low as 10(-12)M synthesized oligonucleotides and 2.67 x 10(2) colony forming unit (CFU)/ml E. coli O157:H7 cells can be detected by the sensor. Linear correlation between frequency change and logarithmic number of bacterial cell concentration was found for E. coli O157:H7 from 2.67 x 10(2) to 2.67 x 10(6)CFU/ml.

A label-free electrochemical impedance immunosensor for rapid detection of Escherichia coli O157:H7 was developed by immobilizing anti-E. coli antibodies onto an indium-tin oxide interdigitated array (IDA) microelectrode. Based on the general electronic equivalent model of an electrochemical cell and the behavior of the IDA microelectrode, an equivalent circuit, consisting of an ohmic resistor of the electrolyte between two electrodes and a double layer capacitor, an electron-transfer resistor, and a Warburg impedance around each electrode, was introduced for interpretation of the impedance components of the IDA microelectrode system. The results showed that the immobilization of antibodies and the binding of E. coli cells to the IDA microelectrode surface increased the electron-transfer resistance, which was directly measured with electrochemical impedance spectroscopy in the presence of [Fe(CN)(6)](3-/4-) as a redox probe. The electron-transfer resistance was correlated with the concentration of E. coli cells in a range from 4.36 x 10(5) to 4.36 x 10(8) cfu/mL with the detection limit of 10(6) cfu/mL.