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Extracellular electron transfer (EET) is key to the success of microbial fuel cells (MFCs). Clostridium sp. often occurs in MFC anode communities, but its ability to perform EET remains controversial. We have employed Clostridium pasteurianum DSM 525 as a biocatalyst in a glycerol-fed MFC, designated MFCDSM. We have also followed the EET of this biocatalyst in the presence of a mediator, namely soluble neutral red (NR), soluble methyl viologen (MV), neutral red film (FNR), or methyl viologen film (FMV). MFCDSM provided power and current densities (j) of 0.39 μW·cm−2 and 2.47 μA·cm−2, respectively, which evidenced that the biocatalyst performs direct electron transfer (DET). Introducing 150.0 µM NR or MV into the MFCDSM improved the current density by 7.0- and 3.7-fold (17.05 and 8.45 μA·cm−2), respectively. After 20 cyclic voltammetry (CV) cycles, the presence of FNR in the MFCDSM anodic chamber provided an almost twofold higher current density (30.76 µA·cm−2) compared to the presence of NR in the MFCDSM. Introducing MV or FMV into the MFCDSM anodic chamber gave practically the same current density after 10 CV cycles. The MFCDSM anodic electrode might interact with FMV weakly than with FNR, so FNR is more promising to enhance C. pasteurianum DSM 525 EET within MFCDSM.

Background and Aim: Feline immunodeficiency virus (FIV) is a retroviral pathogen globally responsible for immunodeficiency disease in cats. However, the current diagnosis based on antibody detection has limitations and can also produce false-positive results. This study aimed to develop a one-pot loop-mediated isothermal amplification (LAMP) process integrated with neutral red (NR-LAMP) assay for detection of FIV proviral DNA. Materials and Methods: We developed a one-pot, gag gene-based NR-LAMP for convenient, rapid, specific, and sensitive colorimetric inspection of FIV proviral DNA. Results: The developed NR-LAMP was capable of amplifying at an optimum temperature of 65°C for 40 min. No cross-amplification was detected between FIV and other feline viruses tested, indicating the high specificity (98.44%) of the novel FIV-LAMP primer. Our NR-LAMP assay has a detection limit of 4.2 × 101 copies/μL. A total of 80 clinical samples with a background of FIV infection were collected and tested using the proposed method. The NR-LAMP assay showed a high sensitivity of 100% compared to conventional polymerase chain reaction assay. Conclusion: These results support the suitability of NR-LAMP as a potential future alternative clinical molecular approach for further use in the diagnosis of FIV-infected cats.

Ruminant livestock industries can support the climate stabilization ambitions of the Paris Agreement through interventions that reduce GHG emissions (predominantly biogenic methane) and sequester carbon in landscapes. This study explored pathways for the Australian red meat industry (grazing, feedlot finishing, and domestic processing) to become climate neutral, whereby the radiative forcing (RF) footprint is plateaued and there is no additional forcing contribution. Emissions timeseries (CO2, N2O, CH4) were compiled for 1990 to 2020 and projected to 2030 under a business-as-usual scenario (including an 18% increase in sheep and 13% increase in beef cattle) and with a range of production system and vegetation management interventions. The RF footprint peaked in 2018 at 7.13 mW/m2 and decreased to 7.07 mW/m2 in 2020. With the future expansion of the herd/flock and under business-as-usual conditions, the RF footprint is projected to increase by 2.8% by 2030. However, with a combination of interventions, production has the potential to increase with a decreasing RF footprint, a condition that can be described as climate neutral. The Australian red meat industry has made an historical contribution to global RF increase. However, with ongoing RF management, it is possible to increase food production within climate-neutral limits.

Measuring pH with an optical sensor requires the immobilization of a chemical recognition phase on a solid surface. Neutral red (NR), an acid base indicator was used to develop two different optical probe configurations. The chemistry of aryl diazonium salts was chosen for the elaboration of this chemical phase, as it enables strong covalent bonds to be established on the surface of metallized glass or metallic surfaces. It also allows the formation of a thick film required to obtain an exploitable spectral response. The surfaces of interest (metallized optical fiber and 316 L stainless-steel mirror) are modelized by flat surfaces (metallized glass plates and 316 L stainless-steel plates). The analytical characterizations carried out (IR, XPS, UV-Visible, and profilometry) show that NR was covalently grafted onto the model surfaces as well as on the surfaces of interest. The supports grafted with NR to develop optical pH probes exhibit spectral changes, particularly the values of pKa, the pH range, and the isosbestic point wavelength. The experimental results show that the optical probe can be used for pH measurements between 4 and 8.

Measuring pH with an optical sensor requires the immobilization of a chemical recognition phase on a solid surface. Neutral red (NR), an acid base indicator was used to develop two different optical probe configurations. The chemistry of aryl diazonium salts was chosen for the elaboration of this chemical phase, as it enables strong covalent bonds to be established on the surface of metallized glass or metallic surfaces. It also allows the formation of a thick film required to obtain an exploitable spectral response. The surfaces of interest (metallized optical fiber and 316 L stainless-steel mirror) are modelized by flat surfaces (metallized glass plates and 316 L stainless-steel plates). The analytical characterizations carried out (IR, XPS, UV-Visible, and profilometry) show that NR was covalently grafted onto the model surfaces as well as on the surfaces of interest. The supports grafted with NR to develop optical pH probes exhibit spectral changes, particularly the values of pKa, the pH range, and the isosbestic point wavelength. The experimental results show that the optical probe can be used for pH measurements between 4 and 8. [ABSTRACT FROM AUTHOR]

Metal-free hydrogen-bonding conductive polymer catalysts (HCPCs) are emerging alternatives to platinum-group-metals (PGMs) for electrocatalytic hydrogen evolution reaction (HER). In this study, electrodes coated with poly-neutral red (PNR) films were fabricated by oxidative chemical vapor deposition (oCVD) and electropolymerization deposition (EPD) to study their activities as HER-electrocatalysts in acidic media. PNR (EPD) exhibited a superior performance, i.e., an overpotential of 194 mV (at 10 mA cm−2) owing to a high exchange current density of 0.711 mA cm−2. The activity is substantiated by spectroelectrochemical measurements and time-dependent density functional theory (TDDFT) calculation unveiling a doubly reduced and doubly protonated PNRH2 as the reaction intermediate for the evolution of H2. It resulted in a first-order kinetic constant of k = 9.98 × 10−4 s−1 to indicate a slow dissociation of PNRH2 to release H2. Large Tafel slope of 176 mV dec−1 also limits the electrocatalytic activity of PNR.

The neutral red uptake (NRU) assay is a cell viability assay that can be used for the assessment of compound-induced cytotoxicity. It is based on the ability of living cells to incorporate neutral red, a weak cationic dye, in lysosomes. The quantification of xenobiotic-induced cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red when compared to cells exposed to corresponding vehicle controls. The NRU assay is mainly used for hazard assessment in in vitro toxicology applications. Hence, this method has been incorporated in regulatory recommendations such as the OECD test guideline TG 432, in which an in vitro 3T3-NRU-phototoxicityassay is described to assess the cytotoxicity of compounds in the presence or absence of UV light.This book chapter describes a detailed protocol to carry out the NRU assay using the human hepatoma cell line HepG2, which is frequently employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetylsalicylic acid is assessed., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: Phototoxicity is an acute photoinduced reaction. The 3T3 neutral red uptake (NRU) phototoxicity test has high sensitivity for the determination of phototoxic substances. To further optimize the method, this study mainly focused on comparing the phototoxicity sensitivity by using the NRU method for BALB/c 3T3, HaCaT, and HDFa cells in vitro. Methods: The NRU method was used to evaluate the phototoxicity of chlorpromazine hydrochloride (CPZ), amiodarone hydrochloride (Amiodar), and L-histidine (L-His) on BALB/c 3T3 cells, HaCaT cells, and HDFa cells. The sensitivity of different cells to ultraviolet (UVA) irradiation in vitro was studied. Results: L-His showed no phototoxicity, but the phototoxicity of CPZ and Amiodar showed different sensitivities among the three kinds of cells. The in vitro phototoxicity evaluation of HaCaT cells is closer to that of primary human fibroblasts. Conclusion: This study provides a reference for cell line selection to optimize the existing in vitro evaluation method of 3T3 NRU phototoxicity.

Multiple sclerosis, neuromyelitis optica, Guillain-Barré syndrome and chronic inflammatory demyelinating polyradiculoneuropathy are representative demyelinating diseases of the central and peripheral nervous system. Remyelination by myelin forming cells is important for functional recovery from the neurological deficits caused in the demyelinating diseases. Lysophosphatidylcholine-induced demyelination in mice is commonly used to identify and study the molecular pathways of demyelination and remyelination. However, detection of focally demyelinated lesions is difficult and usually requires sectioning of demyelinated lesions in tissues for microscopic analysis. In this review, we describe the development and application of a novel vital staining method for labeling demyelinated lesions using intraperitoneal injection of neutral red (NR) dye. NR labeling reduces the time and effort required to search for demyelinated lesions in tissues, and facilitates electron microscopic analysis of myelin structures. NR labeling also has the potential to contribute to the elucidation of pathologies in the central and peripheral nervous system and assist with identification of drug candidates that promote remyelination., Competing Interests: VThe authors declare no conflicts of interest., (2024 The Japan Society of Histochemistry and Cytochemistry.)

Deep eutectic solvents (DES), which have low toxicity and are low cost, biodegradable, and easily synthesized, were used for the extraction of neutral red (NR) dye before its spectrophotometric analysis. DES, containing choline chloride as a hydrogen bond acceptor and phenol as a hydrogen bond donor with a molar ratio of 1:2, was used for the extraction of NR dye from aqueous media. The possible interaction of different DESs with NR was studied using density functional theory (DFT) calculations. Experimentally, a UV-visible spectrophotometer was used for the quantitative analysis. The most important parameters affecting method performance, such as pH, extraction temperature, DES type, its volume, THF volume, sonication time, and centrifugation time, were optimized. The developed method provides exceptional sensitivity in terms of LOD and LOQ, which were 2.2 and 7.3 µg/L respectively. The relative standard deviation was 1.35−1.5% (n = 10), and the pre-concentration factor was 40. The method was found to be linear in the range of 2−300 µg/L (R2 = 0.9967). The method was successfully used for the determination of NR in wastewater samples. Finally, the DES-based method presents operational simplicity, high sensitivity, and rapid determination (<5 min) compared with other analytical procedures.

The three-layer electrode consisting of the inner, middle, and outer layers of polythiophene (PTh), polyaniline (PANI), and poly(neutral red) (PNR), respectively, was developed, characterized, and tested as a potentiometric sensor for citrates. The spectroscopic and morphological findings based on Raman spectroscopy and scanning electron microscopy, respectively, demonstrated the consecutive formation of individual polymeric layers derived from PTh, PANI, and PNR in the multilayer system. The sharper and narrower peak profiles of PNR in the case of the three-layer system revealed a more organized structure than for the PNR layer alone. The PNR layer in such a novel arrangement shows the highest selectivity towards citrates among the tested carboxylates. Simultaneously, the unwanted influence of the underlying Pt surface is eliminated. The potentiometric characteristics of the proposed potentiometric sensor were examined at the detection of citrates in the real-world samples, compared with results for PNR simple electrode, and corresponded with the reference capillary electrophoresis and literature-based spectrophotometric method.