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Membrane potential (MP) changes can provide a simple readout of bacterial functional and metabolic state or stress levels. While several optical methods exist for measuring fast changes in MP in excitable cells, there is a dearth of such methods for absolute and precise measurements of steady-state membrane potentials (MPs) in bacterial cells. Conventional electrode-based methods for the measurement of MP are not suitable for calibrating optical methods in small bacterial cells. While optical measurement based on Nernstian indicators have been successfully used, they do not provide absolute or precise quantification of MP or its changes. We present a novel, calibrated MP recording approach to address this gap. In this study, we used a fluorescence lifetime-based approach to obtain a single-cell resolved distribution of the membrane potential and its changes upon extracellular chemical perturbation in a population of bacterial cells for the first time. Our method is based on (i) a unique VoltageFluor (VF) optical transducer, whose fluorescence lifetime varies as a function of MP via photoinduced electron transfer (PeT) and (ii) a quantitative phasor-FLIM analysis for high-throughput readout. This method allows MP changes to be easily visualized, recorded and quantified. By artificially modulating potassium concentration gradients across the membrane using an ionophore, we have obtained a Bacillus subtilis-specific MP versus VF lifetime calibration and estimated the MP for unperturbed B. subtilis cells to be -65 mV (in MSgg), -127 mV (in M9) and that for chemically depolarized cells as -14 mV (in MSgg). We observed a population level MP heterogeneity of ∼6-10 mV indicating a considerable degree of diversity of physiological and metabolic states among individual cells. Our work paves the way for deeper insights into bacterial electrophysiology and bioelectricity research.

HYPOTHESIS: In this communication, we test the hypothesis that sulfotransferase 1C2 (SULT1C2, UniProt accession no. Q9WUW8) can modulate mitochondrial respiration by increasing state-III respiration. METHODS AND RESULTS: Using freshly isolated mitochondria, the addition of SULT1C2 and 3-phosphoadenosine 5 phosphosulfate (PAPS) results in an increased maximal respiratory capacity in response to the addition of succinate, ADP, and rotenone. Lipidomics and thin-layer chromatography of mitochondria treated with SULT1C2 and PAPS showed an increase in the level of cholesterol sulfate. Notably, adding cholesterol sulfate at nanomolar concentration to freshly isolated mitochondria also increases maximal respiratory capacity. In vivo studies utilizing gene delivery of SULT1C2 expression plasmids to kidneys result in increased mitochondrial membrane potential and confer resistance to ischemia/reperfusion injury. Mitochondria isolated from gene-transduced kidneys have elevated state-III respiration as compared with controls, thereby recapitulating results obtained with mitochondrial fractions treated with SULT1C2 and PAPS. CONCLUSION: SULT1C2 increases mitochondrial respiratory capacity by modifying cholesterol, resulting in increased membrane potential and maximal respiratory capacity. This finding uncovers a unique role of SULT1C2 in cellular physiology and extends the role of sulfotransferases in modulating cellular metabolism.

Magnetogenetics was developed to remotely control genetically targeted neurons. A variant of magnetogenetics uses magnetic fields to activate transient receptor potential vanilloid (TRPV) channels when coupled with ferritin. Stimulation with static or RF magnetic fields of neurons expressing these channels induces Ca2+ transients and modulates behavior. However, the validity of ferritin-based magnetogenetics has been questioned due to controversies surrounding the underlying mechanisms and deficits in reproducibility. Here, we validated the magnetogenetic approach Ferritin-iron Redistribution to Ion Channels (FeRIC) using electrophysiological (Ephys) and imaging techniques. Previously, interference from RF stimulation rendered patch-clamp recordings inaccessible for magnetogenetics. We solved this limitation for FeRIC, and we studied the bioelectrical properties of neurons expressing TRPV4 (nonselective cation channel) and transmembrane member 16A (TMEM16A; chloride-permeable channel) coupled to ferritin (FeRIC channels) under RF stimulation. We used cultured neurons obtained from the rat hippocampus of either sex. We show that RF decreases the membrane resistance (Rm) and depolarizes the membrane potential in neurons expressing TRPV4FeRIC RF does not directly trigger action potential firing but increases the neuronal basal spiking frequency. In neurons expressing TMEM16AFeRIC, RF decreases the Rm, hyperpolarizes the membrane potential, and decreases the spiking frequency. Additionally, we corroborated the previously described biochemical mechanism responsible for RF-induced activation of ferritin-coupled ion channels. We solved an enduring problem for ferritin-based magnetogenetics, obtaining direct Ephys evidence of RF-induced activation of ferritin-coupled ion channels. We found that RF does not yield instantaneous changes in neuronal membrane potentials. Instead, RF produces responses that are long-lasting and moderate, but effective in controlling the bioelectrical properties of neurons.

The demand for multicore processors has significantly increased with the rapid advancement in technology. Effective thermal managements for multicore processors are crucial for extending their operational lifespan. This paper estimates the core temperature of multi-core processors based on the membrane potential of the spiking neural network (SNN). This research collects hardware information of multi-core processors on real machines and estimates the temperature of the processor based on spiking neural network. We use proxy gradients to implement network updates and propose an AMP method to represent temperature. Experimental results reveal that the MAPE between estimated and actual processor temperatures averages almost around 7%, demonstrating the method’s high generalizability across various continuous value regression problems. In addition, the code for collecting the dataset used in this paper will be publicly available. [ABSTRACT FROM AUTHOR]

Background: Ewing's sarcoma is a childhood bone and soft tissue cancer with poor prognosis. Treatment outcomes for Ewing's sarcoma patients have improved only modestly over the past decades, making the development of new treatment strategies paramount. In this study, the combined targeting of ribonucleotide reductase (RNR) and WEE1 was explored for its effectiveness against Ewing's sarcoma cells. Methods: The RNR inhibitor triapine and the WEE1 inhibitors adavosertib and ZN-c3 were tested in p53 wild-type and p53 mutant Ewing's sarcoma cells. The combination of adavosertib with the PARP inhibitors olaparib and veliparib was tested for comparison. Combinatorial effects were determined by flow cytometric analyses of cell death, loss of mitochondrial membrane potential and DNA fragmentation as well as by caspase 3/7 activity assay, immunoblotting and real-time RT-PCR. The drug interactions were assessed using combination index analysis. Results: RNR and WEE1 inhibitors were weakly to moderately effective on their own, but highly effective in combination. The combination treatments were similarly effective in p53 wild-type and p53 mutant cells. They synergistically induced cell death and cooperated to elicit mitochondrial membrane potential decay, to activate caspase 3/7 and to trigger DNA fragmentation, evidencing the induction of the apoptotic cell death cascade. They also cooperated to boost CHK1 phosphorylation, indicating augmented replication stress after combination treatment. In comparison, the combination of adavosertib with PARP inhibitors produced weaker synergistic effects. Conclusion: Our findings show that combined inhibition of RNR and WEE1 was effective against Ewing's sarcoma in vitro. They thus provide a rationale for the evaluation of the potential of combined targeting of RNR and WEE1 in Ewing's sarcoma in vivo. [ABSTRACT FROM AUTHOR]

• Upon critical examination of the arguments in these two Lee papers, we find key weaknesses and fallacies. • Upon close examination of the literature cited by Lee, we find. • strong support for the GHK equation; • published measurements that contradict TELC predictions; and. • no convincing support for the TELC hypothesis. In two recent papers (Curr Trends Neurol 17: 83–98, 2023; J Neurophysiol 124: 1029–1044, 2020), James Lee has argued that his T ransmembrane E lectrostatically- L ocalized C ations (TELC) hypothesis offers a model of neuron transmembrane potentials that is superior to Hodgkin-Huxley classic cable theory and the Goldman-Hodgkin-Katz (GHK) equation. Here we examine critically the arguments in these papers, finding key weaknesses and fallacies. We also examine closely the literature cited by Lee, and find (i) strong support for the GHK equation; (ii) published measurements that contradict TELC predictions; and (iii) no convincing support for the TELC hypothesis. [ABSTRACT FROM AUTHOR]

Ischaemia-reperfusion (IR) damage is an inevitable adverse effect of liver surgery. Recent research has found that IR damage is involved in severe mitochondrial dysfunction. Mitochondrial biosynthesis and dynamics control mitochondrial mass, distribution, and function. Sirtuin 3 (SIRT3) is widely known for preserving health and functionality of mitochondria. DEX has been proven to alleviate liver damage through antioxidant and anti-apoptotic pathways. But it's unclear how DEX protects mitochondria at this time. In this research, the mechanism behind the protective benefits of DEX was examined using the rat liver IR model and the rat liver cells (BRL-3 A) hypoxia reoxygenation (HR) model. We discovered that DEX treatment restored mitochondrial membrane potential, promoted ATP production, prevented oxidative stress, and decreased apoptosis in BRL-3 A cells. Furthermore, HR damage increased mitochondrial fission while decreasing mitochondrial fusion and biogenesis in BRL-3 A cells, which DEX partially corrected. The benefits of DEX on mitochondrial protection were reversed after addition of SR-18,292. Additionally, DEX showed the ability to enhance SIRT3 expression, and after cells were transfected with SIRT3 siRNA, DEX's effects on mitochondria were partially prevented. Similarly, in the rat model, DEX alleviating liver histopathological injury and oxidative stress. DEX inhibited IR-induced mitochondrial damage through improving ETC complex I- IV activities and ATP content, reducing apoptosis, controlling mitochondrial quality, and upregulating the expression of SIRT3. Additionally, our research shows that DEX's ability to protect the liver against IR damage is mediated by the modulation of mitochondrial quality control. Overall, the modification of SIRT3 activity could be responsible for this outcome. [ABSTRACT FROM AUTHOR]

An electrically activable ZnS nanochip for controlled delivery of H 2 S and Zn2+ was developed for gas-ionic-immunotherapy. This work presented a ZnS nanochip for the mobilization of CD8+ T lymphocytes infiltration, reversion of the immunosuppressive tumor microenvironment, which may shed light on the development of electrically activable nanoplatforms for enhanced immunotherapy. [Display omitted] Excess intracellular H 2 S induces destructive mitochondrial toxicity, while overload of Zn2+ results in cell pyroptosis and potentiates the tumor immunogenicity for immunotherapy. However, the precise delivery of both therapeutics remains a great challenge. Herein, an electrically activable ZnS nanochip for the controlled release of H 2 S and Zn2+ was developed for enhanced gas-ionic-immunotherapy (GIIT). Under an electric field, a locality with particularly high concentrations of H 2 S and Zn2+ was established by the voltage-controlled degradation of the ZnS nanoparticles (NPs). Consequently, the ZnS nanochip-mediated gas-ionic therapy (GIT) resulted in mitochondrial membrane potential depolarization, energy generation inhibition, and oxidative stress imbalance in tumor cells. Interestingly, the cyclic guanosine monophosphate-adenosine monophosphate synthase-stimulator of interferon genes (cGAS-STING) signaling pathway was activated due to the mitochondrial destruction. Moreover, the released Zn2+ resulted in the increase of the intracellular Zn levels and cell pyroptosis, which enhanced the immunogenicity via the release of damage-associated molecular patterns (DAMPs). In vitro and in vivo studies revealed that the ZnS nanochip-based GIT effectively eliminated the tumors under an electric field and mobilized the cytotoxic T lymphocytes for immunotherapy. The combination with αCTLA-4 further promoted the adaptive immune response and inhibited tumor metastasis and long-term tumor recurrence. This work presented an electrically activable ZnS nanochip for combined immunotherapy, which might inspire the development of electric stimulation therapy. [ABSTRACT FROM AUTHOR]

DYT1 dystonia is an inherited early-onset generalized dystonia characterized by sustained muscle contractions causing abnormal, repetitive movements or postures. Most DYT1 patients have a heterozygous trinucleotide GAG deletion (ΔGAG) in DYT1/TOR1A, coding for torsinA. Dyt1 heterozygous ΔGAG knock-in (KI) mice or global KI mice show motor deficits and abnormal Purkinje cell firing. However, Purkinje cell-specific heterozygous ΔGAG conditional KI mice (Pcp2-KI) show improved motor performance, reduced sensory-evoked brain activation in the striatum and midbrain, and reduced functional connectivity of the striatum with the anterior medulla. Whether Pcp2-KI mice show similar abnormal Purkinje cell firing as the global KI mice, suggesting a cell-autonomous effect causes the abnormal Purkinje cell firing in the global KI mice, is unknown. We used acute cerebellar slice recording in Pcp2-KI mice to address this issue. The Pcp2-KI mice exhibited no changes in spontaneous firing and intrinsic excitability compared to the control mice. While membrane properties were largely unchanged, the resting membrane potential was slightly hyperpolarized, which was associated with decreased baseline excitability. Our results suggest that the abnormal Purkinje cell firing in the global KI mice was not cell-autonomous and was caused by physiological changes elsewhere in the brain circuits. Our results also contribute to the ongoing research of how basal ganglia and cerebellum interact to influence motor control in normal states and movement disorders. [ABSTRACT FROM AUTHOR]

The Shaker/Kv1 subfamily of voltage-gated potassium (K+) channels is essential for modulating membrane excitability. Their loss results in prolonged depolarization and excessive calcium influx. These channels have also been implicated in a variety of other cellular processes, but the underlying mechanisms remain poorly understood. Through comprehensive screening of K+ channel mutants in C. elegans, we discovered that shk-1 mutants are highly susceptible to bacterial pathogen infection and oxidative stress. This vulnerability is associated with reduced glycogen levels and substantial mitochondrial dysfunction, including decreased ATP production and dysregulated mitochondrial membrane potential under stress conditions. SHK-1 is predominantly expressed and functions in body wall muscle to maintain glycogen storage and mitochondrial homeostasis. RNA-sequencing data reveal that shk-1 mutants have decreased expression of a set of cation-transporting ATPases (CATP), which are crucial for maintaining electrochemical gradients. Intriguingly, overexpressing catp-3, but not other catp genes, restores the depolarization of mitochondrial membrane potential under stress and enhances stress tolerance in shk-1 mutants. This finding suggests that increased catp-3 levels may help restore electrochemical gradients disrupted by shk-1 deficiency, thereby rescuing the phenotypes observed in shk-1 mutants. Overall, our findings highlight a critical role for SHK-1 in maintaining stress tolerance by regulating glycogen storage, mitochondrial homeostasis, and gene expression. They also provide insights into how Shaker/Kv1 channels participate in a broad range of cellular processes. Author summary: Exposure to environmental stimuli, such as bacterial pathogens, increases the production of reactive oxygen species (ROS), leading to oxidative stress and widespread cellular damage. Cells employ various defense mechanisms to counteract ROS-induced damages and maintain cellular homeostasis. In this study, we found that the Shaker/Kv1 channel SHK-1, typically known for its regulation of membrane excitability, also helps protect C. elegans against bacterial infections and oxidative stress. Animals lacking shk-1 are highly sensitive to pathogens and prooxidant exposure, and show defects in gene expression, glycogen storage, and mitochondrial function. Notably, shk-1 mutants have reduced expression of catp-3, encoding one of cation-transporting ATPases that are critical for maintaining ion balance. Restoring catp-3 expression rescues the defects observed in shk-1-deficient animals, suggesting that the loss of SHK-1 likely disturbs ion homeostasis, which can be mitigated by increased levels of catp-3. These findings offer a new perspective on the fundamental mechanism by which SHK-1 regulates diverse cellular functions. Moreover, our observations may have implications for how Kv1.3 is involved in various physiological processes in mammals. [ABSTRACT FROM AUTHOR]

IAA is a by-product of the water disinfection process and has been found to be neurotoxic. However, the role and mechanism of IAA neurotoxicity remain unclear. In this review, we comprehensively discuss the neurotoxic effects and mechanisms of IAA from the molecular level, cellular level and neurological manifestations. At the molecular level, IAA causes neurotoxicity by reducing mitochondrial membrane potential, aggravating oxidative stress and DNA damage. At the cellular level, IAA causes neurotoxicity by inducing BBB disruption, neuroinflammation, and apoptosis. In neurological manifestations, IAA can lead to neurotransmitter disorders, neurodevelopment dysfunction, and even neurodegenerative diseases. Taken together, our review provides insights into the mechanisms of IAA neurotoxicity that will contribute to future studies of IAA neurotoxicity and its protective strategies. [ABSTRACT FROM AUTHOR]

Parkinson's disease is a neurodegenerative disorder characterized by cardinal motor symptoms resulting from the loss of dopaminergic neurons in the substantia nigra pars compacta. Although current medications may alleviate its symptoms, Parkinson's disease remains incurable. Astaxanthin is an antioxidant and anti-inflammatory agent; however, its high susceptibility to oxidative degradation and low aqueous solubility limit its therapeutic efficacy. This study aimed to improve the pharmaceutical properties and neuroprotective effects of astaxanthin for Parkinson's disease treatment by using lactoferrin-conjugated astaxanthin-loaded liposomes (Lf-ASX-LPs). We successfully formulated Lf-ASX-LPs with high encapsulation efficiency (97.6%) and favorable physical characteristics (diameter: 109.8 ± 1.1 nm; polydispersity index: 0.18 ± 0.01; zeta potential: − 9.5 ± 1.1 mV). Lf-functionalized liposomes demonstrated enhanced cellular uptake and permeation in a Transwell® study, showing a 16.7-fold higher internalization in SH-SY5Y cells over 24 h than those without Lf conjugation. Additionally, Lf functionalization enhanced brain penetration ability, as demonstrated by a biodistribution study using nude mice, compared to LPs without Lf conjugation. In vitro, Lf-ASX-LPs reduced cell loss by 20.1% and oxygen species by 30.0%, ameliorated the reduction in mitochondrial membrane potential under 1-methyl-4-phenylpyridinium-induced toxicity by 40.1%, and reduced extracellular nitric oxide levels under lipopolysaccharide-induced toxicity by 32.0%, indicating cytoprotective and antioxidant effects. In a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease mouse model, treatment with Lf-ASX-LPs resulted in 5.0-, 3.1-, and 5.6-fold increases in dopamine levels, TH+ fiber density, and TH+ neurons, respectively, restoring dopaminergic neuron damage. The developed formulation also alleviated behavioral impairment and neuroinflammation, reducing astrocyte and microglial activation in the striatum and substantia nigra of the MPTP-treated animals. Thus, our formulation of Lf-ASX-LPs represents a promising strategy for providing neuroprotection and retarding Parkinson's disease progression. [ABSTRACT FROM AUTHOR]

The correction notice in Scientific Reports addresses errors in the original article regarding the figure assembly process. The correction includes updated figures showing the correct images for specific time points related to the effects of amifostine on normal tissue radioprotection. The authors, including Michael I. Koukourakis and Adrian L. Harris, conducted experiments on mouse liver cells and hepatoma cells to analyze the impact of amifostine on anaerobic metabolism and mitochondrial function. The correction ensures the accuracy of the research findings presented in the article. [Extracted from the article]

To elucidate the molecular function of SHORT AND SWOLLEN ROOT1 (SSR1), we screened for suppressors of the ssr1-2 (sus) was performed and identified over a dozen candidates with varying degrees of root growth restoration. Among these, the two most effective suppressors, sus1 and sus2, resulted from G87D and T55M single amino acid substitutions in HSCA2 (At5g09590) and ISU1 (At4g22220), both crucial components of the mitochondrial iron-sulfur (Fe-S) cluster assembly machinery. SSR1 displayed a robust cochaperone-like activity and interacted with HSCA2 and ISU1, facilitating the binding of HSCA2 to ISU1. In comparison to the wild-type plants, ssr1-2 mutants displayed increased iron accumulation in root tips and altered expression of genes responsive to iron deficiency. Additionally, the enzymatic activities of several iron-sulfur proteins and the mitochondrial membrane potential were reduced in ssr1-2 mutants. Interestingly, SSR1 appears to be exclusive to plant lineages and is induced by environmental stresses. Although HSCA2G87D and ISU1T55M can effectively compensate for the phenotypes associated with SSR1 deficiency under favorable conditions, their compensatory effects are significantly diminished under stress. Collectively, SSR1 represents a new and significant component of the mitochondrial Fe-S cluster assembly (ISC) machinery. It may also confer adaptive advantages on plant ISC machinery in response to environmental stress. Author summary: Iron-sulfur (Fe-S) clusters are crucial components found in many proteins that play essential roles in various biological processes. The machinery responsible for making these clusters in mitochondria, known as ISC biosynthesis, has its origins in bacteria and remained largely unchanged through evolution. However, understanding of specific regulators that control Fe-S cluster production in plants is still limited. In this study, we identified a unique protein in plants, SSR1, which acts as a cochaperone. SSR1 facilitates the interaction between the two critical proteins, HSCA2 and ISU1, a necessary process for the release of Fe-S clusters from their scaffold. We also showed that SSR1 has evolved alongside the ISC biosynthetic machinery and that mutations in HSCA2 and ISU1 can compensate for its absence. This highlights the synergistic relationship between SSR1 and other components of the ISC machinery. Overall, this research uncovers a novel component of the ISC biosynthetic system and shows how it varies between plants, animals, and microorganisms. [ABSTRACT FROM AUTHOR]

Introduction: The vast majority of cellular studies on mammalian vestibular hair cells have been carried out in rodent models due in part to the inaccessibility of human inner ear organs and reports describing electrophysiological recordings from human inner ear sensory hair cells are scarce. Here, we obtained freshly harvested vestibular neuroepithelia from adult translabyrinthine surgical patients to obtain electrophysiological recordings from human hair cells. Methods: Whole cell patch clamp recordings were performed on hair cells mechanically isolated from human cristae to characterize voltage-dependent and pharmacological properties of membrane currents. Hair cells were classified as type I or type II according to morphological characteristics and/or their electrophysiological properties. Results: Type I hair cells exhibited low voltage-activated K+ currents (IKLV) at membrane potentials around the mean resting membrane potential (-63 mV) and large slowly activating outward K+ currents in response to depolarizing voltage steps. Recordings from type II hair cells revealed delayed rectifier type outward K+ currents that activated above the average resting potential of -55 mV and often showed some inactivation at more depolarized potentials. Perfusion with the K+ channel blocker 4-aminopyridine (1 mM) substantially reduced outward current in both hair cell types. Additionally, extracellular application of 8-bromo-cGMP inhibited IKLV in human crista type I hair cells suggesting modulation via a nitric oxide/cGMP mechanism. A slow hyperpolarization-activated current (Ih) was observed in some hair cells in response to membrane hyperpolarization below -100 mV. Discussion: In summary, whole cell recordings from isolated human hair cells revealed ionic currents that strongly resemble mature current phenotypes previously described in hair cells from rodent vestibular epithelia. Rapid access to surgically obtained adult human vestibular neuroepithelia allows translational studies crucial for improved understanding of human peripheral vestibular function. [ABSTRACT FROM AUTHOR]

Background: HECT domain and Ankyrin repeat Containing E3 ubiquitin-protein ligase 1 (HACE1) has been found to be associated with mitochondrial protection. Mitochondrial damage is a critical contributor to myocardial ischemia–reperfusion injury (I/RI). However, little is known about the role of HACE1 in the pathogenesis of myocardial I/RI. Methods: Male C57BL6 mice with HACE1 knockout (KO) were subjected to 30 min of ischemia via ligation of the left anterior descending artery, followed by 0, 2, 6, or 24 h of reperfusion. The mice were evaluated for myocardial histopathological injury, serum troponin I (cTnI) levels, oxidative stress injury, apoptosis and cardiac function. Prior to ischemia, Mdivi-1(1.2 mg/kg) or vehicle was administered. Results: The study revealed that increased expression of HACE1 was associated with myocardial ischemia/reperfusion injury (I/RI), and that knockout of HACE1 resulted in more severe myocardial damage and cardiac dysfunction during I/R(P < 0.05). The HACE1 knockout group exhibited higher levels of malondialdehyde (MDA), greater mitochondrial fission, and dissipation of mitochondrial membrane potential (MMP), leading to more apoptosis and severe cardiac dysfunction compared to the wild-type I/R group(P < 0.05). On the other hand, HACE1 knockout further reduced superoxide dismutase (SOD) activity in the myocardium(P < 0.05), further supporting the findings. However, the adverse effects were almost completely eliminated by pharmacological blockade of the dynamin-related protein 1 (Drp1) inhibitor, Mdivi-1, which inhibits mitochondrial fission during cardiac I/R(P < 0.05). Conclusion: Collectively, our data show that myocardial I/RI is associated with HACE1 downregulation and Drp1 activation, causing cardiomyocytes to undergo cell death. Therefore, HACE1 could be a promising therapeutic target for the treatment of myocardial I/RI. [ABSTRACT FROM AUTHOR]

Optogenetic stimulation combined with optical mapping of membrane potential (Vm) and calcium transients (CaT) is a powerful electrophysiological tool. We developed a novel experimental platform in which tissue is stimulated optogenetically while Vm and CaT are imaged simultaneously. The Vm indicator is an organic dye, while the CaT indicator is genetically encoded. We used cardiac spheroids containing cardiomyocytes and fibroblasts differentiated from human induced pluripotent stem cells as model tissue. The spheroids were genetically encoded with an optogenetic actuator, CheRiff, and the calcium indicator jRCaMP1b. The Vm indicator was the organic dye RH237. CheRiff was excited using blue light (450 nm), and both RH237 and jRCaMP1b were excited using a single band of green light (either 525–575 nm or 558–575 nm). Fluorescence emission was split and imaged by two cameras (CaT: 595–665 nm; Vm: >700 nm). The spheroids were successfully stimulated optogenetically and Vm and CaT were recorded simultaneously without cross-talk using both excitation light bands. The 525–575 nm band produced higher signal-to-noise ratios than the 558–575 nm band, but caused a slight increase in tissue excitability because of CheRiff activation. The optogenetic actuator and CaT indicator are genetically encoded and can be expressed in engineered tissue constructs. In contrast, the Vm indicator is an organic dye that can stain any tissue. This system is well-suited for studying coupling between engineered tissue grafts and host tissue because the two tissue types can be stimulated independently, and tissue activation can be unambiguously attributed to either graft or host. [Display omitted] • Spectral congestion is a serious issue in "all-optical" electrophysiology. • Developed a novel system combining optogenetics and simultaneous Vm and CaT mapping. • CheRiff for optogenetics, jRCaMP1b for CaT, and organic dye RH237 for Vm recordings. • Demonstrated the system in engineered cardiac tissue without Vm/CaT cross-talk. • Applicable for studying bidirectional electrical coupling in cell transplantation. [ABSTRACT FROM AUTHOR]

In vitro maturation (IVM) plays a critical role in embryo production. However, the quality of IVM oocytes often suffers from oxidative stress due to the excessive accumulation of ROS. Equol, a metabolite of soybean flavonoids, exhibits potent antioxidant activity. This study investigated the effects of equol on porcine oocyte IVM. Our findings showed that treatment with 5 μM equol significantly enhanced cumulus cell expansion and the first polar body extrusion in porcine oocytes. Moreover, equol also improved the subsequent embryonic development capacity of the oocytes after parthenogenetic activation. Additionally, equol improved mitochondrial function by increasing mitochondrial content, membrane potential, and ATP levels, while promoting lipid droplet accumulation in oocytes. Equol also reduced DNA damage and early apoptosis, with an associated upregulation of BCL2 and downregulation of BAX expression. Notably, equol decreased ROS levels, likely through activation of the NRF2/KEAP1 antioxidant pathway, leading to increased expression of HO-1 , CAT , GPX1 , and SOD. In conclusion, equol improves porcine oocyte IVM by mitigating oxidative stress via activation of the NRF2/KEAP1 pathway, offering a potential strategy for optimizing the IVM system in porcine oocytes. [Display omitted] [ABSTRACT FROM AUTHOR]

The NLRP3 inflammasome plays a pivotal role in the progression of inflammatory diseases. Mitochondrial damage, oxidative stress and mitochondrial DNA (mtDNA) leak are the key upstream factors for NLRP3 inflammasome activation. Nepetin (Nep), a naturally occurring flavonoid found with anti-inflammatory properties; however, whether it can affect the NLRP3 inflammasome activation and its precise anti-inflammatory mechanism remains unclear. In this study, we demonstrated that Nep enhances PINK1-mediated ubiquitin phosphorylation, which promotes mitophagy and subsequently inhibits NLRP3 inflammasome activation and pyroptosis in macrophages. The administration of Nep to macrophages alleviated of mitochondrial damage, reduced mitochondrial superoxide production, restored mitochondrial membrane potential and prevented the mtDNA leakage. These findings provide compelling evidence for the antioxidant effect of Nep. Furthermore, the pivotal function of mitophagy in the NLRP3 inflammasome inhibitory impact of Nep was substantiated through the utilisation of mitophagy inhibitors and siRNA techniques. Notably, Nep increased survival and reduced organ damage in mice with systemic inflammation by inhibiting NLRP3 inflammasome activation. In addition, Nep suppressed NLRP3 inflammasome activation in obese mice, which led to reduced white adipose and liver inflammation, thereby ameliorating insulin resistance. In conclusion, our findings suggest that Nep is a potent NLRP3 inflammasome inhibitor and a promising candidate for the development of anti-inflammatory therapies. [Display omitted] • Nep was identified as a potent NLRP3 inflammasome inhibitor in vivo and in vitro. • Nep exerts antioxidant and mitochondrial protection through mitophagy. • Nep is a PINK1/Parkin pathway activator, enhances mitophagy and exerts anti-NLRP3 inflammasome inhibitory effect. [ABSTRACT FROM AUTHOR]

Chloroquine (CQ), an autophagy antagonist, has been recently explored as a repurposable medicine for cancer; however the exact mechanism of its action is still not known. The present study investigated the effect of CQ on colorectal cancer cells to elucidate the underlying molecular mechanisms. We report for the first time that CQ suppresses hypoxia-induced growth and survival of HCT-116 cells by reducing glycolytic capacity and NAD+ production through inhibition of PDK1. Furthermore, in silico and in vitro studies revealed that CQ induces structural alteration in the PDK1 protein, leading to its destabilization and promotes its enhanced degradation by proteases. This degradation is in turn inhibited by the MG-132 protease inhibitor. Moreover, CQ-induced suppression of PDK1 results in mitochondrial damage through excessive production of ROS, as reflected by the reduction in mitochondrial membrane potential, which in turn triggers apoptosis through PARP cleavage and Caspase activation. These findings advocate CQ as a promising repurposable chemotherapeutic for colorectal cancer and a novel inhibitor of PDK1. In absence of Chloroquine- Active PDK1 inhibits activity of pyruvate dehydrogenase complex (PDHC) promoting Warburg effect, which inhibits ROS generation and stimulates cell survival and growth. In presence of Chloroquine – CQ-induced destabilization of PDK1 promotes its degradation, resulting in increased mitochondrial oxidative phosphorylation (OXPHOs) thereby increasing ROS generation and cell death. [Display omitted] • CQ suppresses proliferation of hypoxic HCT-116 cells through the inhibition of anaerobic glycolysis. • This effect of CQ is facilitated by its targeting of PDK1, a kinase crucial for glycolytic reprogramming during hypoxa. • CQ directly binds to PDK1, affecting its overall conformation, stability, and bioavailability. • CQ-mediated loss of PDK1 triggers ROS-dependent apoptosis in HCT-116 cells by reducing mitochondrial activity. [ABSTRACT FROM AUTHOR]

The main therapeutic strategy for the treatment of patients with toxic liver failure is the elimination of the toxic agent in combination with the targeted mitigation of pathological processes that have been initiated due to the toxicant. In the current research we evaluated the strategy of metabolic supplementation to improve mitochondrial bioenergetics during acute liver intoxication. In our study, we have shown that acute CCl4-induced intoxication negatively affects Complex I (in the presence of glutamate-malate as energy substrates) based respiration, generation of mitochondrial membrane potential (ΔΨm), mitochondrial NAD(P)H pool and NADH redox index, mitochondrial calcium retention capacity (CRC) and structure and functions of the liver. Boosting of mitochondrial bioenergetics through the complex II, using succinate as metabolic substrate in vitro, significantly improved mitochondrial respiration and generation of ΔΨm, but not mitochondrial CRC. Co-application of rotenone along with succinate, to prevent possible reverse electron flow, didn't show significant differences compared to the effects of succinate alone. Treatment of animals with acute liver failure, using a metabolic supplement containing succinate, inosine, methionine and nicotinamide improved Complex I based respiration, generation of ΔΨm, mitochondrial NAD(P)H pool and NADH redox index, mitochondrial CRC and slightly decreased the level of oxidative stress. These changes resulted in averting destructive and dystrophic changes in the structure of rat liver tissue caused by CCl4 intoxication, concomitantly enhancing hepatic functionality. Thus, we propose that metabolic supplementation targeting complex II could serve as a potential adjunctive therapy in the management of acute liver intoxication. [ABSTRACT FROM AUTHOR]

A convenient total synthesis of the imidazo[1,2-f]phenanthridine-type Amaryllidaceae alkaloid zephycandidine A (3) was developed, which further allowed us to perform modifications of substituents on benzenoid ring A and imidazole ring D. The biological activities of all synthesized compounds were evaluated, and it was reported that activities against cancer cells of the parent alkaloid were poorly reproducible, while the closely related analogue THK-121 (11) showed a strong inhibitory effect on proliferation. Additionally, our novel analogue significantly induced cell death via the intrinsic apoptosis pathway, evident by the loss of mitochondrial membrane potential, increased mitochondrial oxidative stress, and disrupted mitochondrial structure in the same cells. At the same time, healthy cells were less affected by the treatment with THK-121 (11), indicating a potential therapeutic margin. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) holds considerable promise for advancing anticancer treatment, owing to its precision and minimally invasive nature. In this study, we successfully synthesized a series of titanium carbide (Ti3C2, TC)/graphitic carbon nitride (g-C3N4, CN) nanocomposite through a synergistic approach combining electron beam irradiation and 2D/2D composite formation. According to the results, 1TC/200-CN (1TC, which TC was 1, referred to the mass ratio; 200-CN, which CN was 200 kGy, referred to the irradiation metering) displayed a 94% degradation rate of methylene blue (10 mg/L) in 100 min. Furthermore, the proliferation rate of CAL-27 cells was suppressed to just 23.3% at a concentration of 320 μg/mL of 1TC/200-CN. Notably, the group treated with this concentration exhibited the largest residual scratch area, accompanied by a notable decrease in mitochondrial membrane potential. These enhanced effects were attributed to the efficient transfer of electron-hole pairs facilitated by the TC/CN composite. Our findings not only contribute to the development of efficient and stable nanocomposites for PDT applications but also provide valuable insights into the utilization of nanomaterials in the biomedical field, thereby paving the way for potential breakthroughs in cancer treatment. [ABSTRACT FROM AUTHOR]

[Display omitted] • Graphene quantum dots (GQDs) bind to the variant surface glycoprotein of T. brucei. • The GQDs interacted with the p18 subunit of T. brucei ATP synthase F 1 and inhibited ATP synthase activity. • GQDs exposure results in a decrease in the proteasome pathway but an increase in the RNA degradation pathway in T. brucei. Trypanosomiasis is one of the main threats to human and animal health in African countries. Trypanosoma brucei can evade the host immune recognition by rapidly altering its variant surface glycoprotein (VSG). The ATP synthase F 1 subunit of the parasite exhibits extremely low similarity to that of its mammalian hosts, hypothetically making it an ideal target for the development of novel therapeutics. Graphene quantum dots (GQDs) were synthesized, and their adhesion to T. brucei surface and internalization was observed microscopically. The activity of ATP synthase and mitochondrial membrane potential of T. brucei were measured after exposure to GQDs. Proteomics, biolayer interferometry, and molecular dynamic simulations were utilized to evaluate the interaction between GQDs with the target proteins. GQDs specifically adhered to the VSG of T. brucei and were conveyed inside the parasite via the VSG internalization pathway. The GQDs promoted intracellular ROS production, interacted with, and inhibited the activity of the p18 subunit of ATP synthase, disrupted parasite mitochondrial membrane potential. Additionally, the GQDs caused a decrease in aminoacyl – tRNA biosynthesis, and upregulated RNA and protein degradation pathways. The findings of this study offer a novel avenue for the target-oriented discovery of anti-trypanosome drugs. [ABSTRACT FROM AUTHOR]

It was previously reported that tyrosine 97 (Y97) of cytochrome c is phosphorylated in cow heart tissue under physiological conditions. Y97 phosphorylation was shown to partially inhibit respiration in vitro in the reaction with purified cytochrome c oxidase. Here, we use phosphomimetic Y97E Cytc to further characterize the functional effects of this modification both in vitro and in cell culture models. In vitro, phosphomimetic Y97E Cytc showed lower activity in the reaction with purified cow heart cytochrome c oxidase (COX), decreased caspase-3 activity, and reduced rate of reduction. Additionally, the phosphomimetic Y97E Cytc tended to be resistant to heme degradation and showed an increased rate of oxidation. Intact mouse Cytc double knockout fibroblasts were transfected with plasmids coding for phosphomimetic Y97E Cytc and other variants. Compared to cells expressing wild-type Cytc, the cells expressing phosphomimetic Y97E Cytc showed reduced respiration, mitochondrial membrane potential, and reactive oxygen species production, and protection from apoptosis. In an oxygen–glucose deprivation/reoxygenation cell culture model of ischemia/reperfusion injury, mitochondrial membrane potential and reactive oxygen species production were decreased. These data show that Cytc phosphorylation controls the overall flux through the electron transport chain by maintaining optimal intermediate ΔΨm potentials for efficient ATP production while minimizing reactive oxygen species production, thus protecting the cell from apoptosis. [ABSTRACT FROM AUTHOR]

Tetraploidy is a condition in which the entire set of chromosomes doubles, most often due to errors during cell division. Tetraploidy can lead to genomic instability and significant consequences, in particular metastasis and treatment failure in tumours, including radiotherapy. The development of new strategies to sensitise these cells to treatment is of great importance. In our study, we investigated the in vitro combination of chemical treatment with the kinase inhibitor SP600125 and irradiation on diploid versus metastatic tetraploid RKO colon cancer clones. We assessed mitochondrial transmembrane potential, cell cycle and subG1 population by flow cytometry and performed clonogenic assays to evaluate cell sensitivity. We found that the combination overcomes irradiation resistance in metastatic tetraploid clones. To identify the main pathway involved in cell sensitivity, we screened the Harvard Medical School KINOMEscan library and performed a gene ontology biological process analysis. We found that the major kinases inhibited by SP600125 were ANKK1, BIKE, IKKA, JNK1, MP2K3, MP2K4, MKNK2, MYLK, PLK4, RPS6KA4(Kin,Dom,1), MYLK4 and TTK, and the pathways involved in clone sensitivity were DNA damage repair, radiation resistance and apoptosis, through JNK pathway inhibition. Finally, our main finding was that combined treatment with SP600125 and radiotherapy reduced the resistance of metastatic tetraploid cells to treatment, essentially by inhibiting the JNK pathway. This result supports a promising anti-cancer strategy to overcome the resistance of tetraploid cancer cells to irradiation. [ABSTRACT FROM AUTHOR]

The use of black soybean (Glycine max L.), an edible crop prevalent in Asia, has attracted attention for its hepatoprotective properties. Notably, the anthocyanin components in black soybean coats have shown potential in inhibiting tumor growth. Here, anthocyanins were extracted from black soybean coats using both microwave and water bath methods. The physicochemical characteristics of black soybean coat anthocyanins (BSCAs) and their biological activities were examined. The results from the MTT and EDU assays demonstrated a dose-dependent inhibitory effect of BSCAs on hepatocellular carcinoma HepG2 cells, while leaving normal cells unaffected. Flow cytometry and mitochondrial membrane potential assays revealed that BSCA treatment induces apoptosis in HepG2 cells. A network pharmacology approach was employed to explore the relationship between hepatocellular carcinoma and the active ingredients of BSCAs, identifying the JAK/STAT signaling pathway as a potential target. Molecular docking studies confirmed the interaction between BSCA components and JAK2/STAT3 targets. Subsequent Western blot and qPCR experiments validated that BSCAs promote apoptosis in HepG2 cells by modulating the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases and cancer are leading global causes of morbidity and mortality, necessitating advances in diagnosis and treatment. Doxorubicin (Doxo), a potent chemotherapy drug, causes long-term heart damage due to cardiotoxicity. Small extracellular vesicles (sEVs) carry bioactive molecules—such as proteins, lipids, and nucleic acids—that can modulate gene expression and signaling pathways in recipient cells, including cardiomyocytes. Through the delivery of cytokines, microRNAs, and growth factors, sEVs can influence cell survival, which plays a critical role in the development of cardiotoxicity. This study investigates the role of sEVs derived from breast cancer cells treated or not with Doxo and their potential to induce cardiomyocyte damage, thereby contributing to cardiotoxicity. We isolated sEVs from MCF-7 cells treated or not to Doxo using ultracentrifugation and characterized them through Nanoparticle Tracking Analysis (NTA), Scanning Electron Microscopy (SEM), and Western Blotting (WB) for the markers CD63, CD81, and TSG101. We analyzed cytokine profiles using a Multiplex Assay and Cytokine Membrane Array. We exposed Guinea pig cardiomyocytes to different concentrations of sEVs. We assessed their viability (MTT assay), shortening, reactive oxygen species (ROS–DHE dye) production, mitochondrial membrane potential (JC-1 dye), and calcium dynamics (FLUO-4 dye). We performed statistical analyses, including t-tests, ANOVA, Cohen's d, and η2 to validate the robustness of the results. Treatment of MCF-7 cells with 0.01 μM Doxorubicin resulted in increased sEVs production, particularly after 48 h of exposure (~1.79 × 108 ± 2.77 × 107 vs. ~5.1 × 107 ± 1.28 × 107 particles/mL, n = 3, p = 0.0019). These sEVs exhibited protein profiles in the 130–25 kDa range and 93–123 nm sizes. They carried cytokines including TNF-α, IL-1β, IL-4, IFN-γ, and IL-10. Exposure of cardiomyocytes to sEVs (0.025 μg/mL to 2.5 μg/mL) from both Doxo-treated and untreated cells significantly reduced cardiomyocyte viability, shortened cell length by up to 20%, increased ROS production, and disrupted calcium homeostasis and mitochondrial membrane potential, indicating severe cellular stress and cardiotoxicity. These findings suggest that Doxo enhances sEVs production from breast cancer cells, which plays a key role in cardiotoxicity through their cytokine cargo. The study highlights the potential of these sEVs as biomarkers for early cardiotoxicity detection and as therapeutic targets to mitigate cardiovascular risks in chemotherapy patients. Future research should focus on understanding the mechanisms by which Doxorubicin-induced sEVs contribute to cardiotoxicity and exploring their diagnostic and therapeutic potential to improve patient safety and outcomes in cancer therapy. [ABSTRACT FROM AUTHOR]

Ferritin stores iron as Fe 3 + after being oxidized in the presence of molecular oxygen. The Fe 2 + ions typically enter into the ferroxidase as hexahydrate, where the oxidation of Fe 2 + to Fe 3 + occurs. The studies predict probable residues involved in the entry of molecular oxygen by sequence and structural comparison with other proteins. This study identified the probable π -H paths for electron transfer and proton translocation in ferritin through structural analysis. The identification of multiple proton translocation paths and the use of hexahydrate iron as substrate by ferritin indicate that during the oxidation of Fe 2 + to Fe 3 + , all the water molecules of iron may be utilized. The resulting byproducts, protons and hydroxyl ions may be one of the sources for the proton gradient required to drive oxidative phosphorylation by electron transport complexes in mitochondria. Also, it may be required to acidify lysosomes and generate nitroperoxy species. Comparative sequence analysis, structural analysis, and molecular dynamics simulation identified the likely route of molecular oxygen entry in ferritin Structural analysis of ferritin invoked several hydrogen bond networks likely involved in proton translocation paths Iron hexahydrate as a substrate instead of simple iron in the ferritin-catalyzed reaction is implicated as a source of protons to maintain the mitochondrial membrane potential and intuitively other cellular needs [ABSTRACT FROM AUTHOR]

Cell swelling is known to be involved in various stages of the growth of plant cells and microorganisms but in mammalian cells how crucial a swollen state is for determining the fate of the cellular proliferation remains unclear. Recent evidence has increased our understanding of how the loss of the cell surface interactions with the extracellular matrix at early mitosis decreases the membrane tension triggering curvature changes in the plasma membrane and the activation of the sodium/hydrogen (Na +/H +) exchanger (NHE1) that drives osmotic swelling. Such a swollen state is temporary, but it is critical to alter essential membrane biophysical parameters that are required to activate Ca2 + channels and modulate the opening of K + channels involved in setting the membrane potential. A decreased membrane potential across the mitotic cell membrane enhances the clustering of Ras proteins involved in the Ca2 + and cytoskeleton-driven events that lead to cell rounding. Changes in the external mechanical and osmotic forces also have an impact on the lipid composition of the plasma membrane during mitosis. [ABSTRACT FROM AUTHOR]

Objective: Triple-Negative Breast Cancer (TNBC), the most challenging subtype of Breast Cancer (BC), currently lacks targeted therapy, presenting a significant therapeutic gap in its management. Tumor Associated Macrophages (TAMs) play a significant role in TNBC progression and could be targeted by repolarizing them from M2 to M1 phenotype. Matairesinol (MAT), a plant lignan, has been shown to exhibit anticancer, anti-inflammatory and immunomodulatory activities. In this study, we explored how MAT-induced repolarization of THP-1-derived M2 macrophages towards the M1 phenotype, which could effectively target the TNBC cell line, MDA-MB-231. Methods: The differential expression of genes in THP-1-derived macrophages at mRNA levels was evaluated by RNAseq assay. An inverted microscope equipped with a CMOS camera was utilized to capture the morphological variations in THP-1 cells and THP-1-derived macrophages. Relative mRNA expression of M1 and M2 specific marker genes was quantified by qRT-PCR. Cell viability and induction of apoptosis were evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1 dye) assays, respectively. Results: MAT reduced the viability of M2a and M2d macrophages and repolarized them to M1 phenotype. Conditioned medium (CM) from MAT-treated M2a and M2d macrophages significantly reduced the viability of TNBC cells by apoptosis. Conclusion: Targeting M2 macrophages is an important strategy to regulate cancer progression. Our study provides evidence that MAT may be a promising drug candidate for developing novel anti-TNBC therapy. However, further studies are warranted to thoroughly elucidate the molecular mechanism of action of MAT and evaluate its therapeutic potential in TNBC in vitro and in vivo models. [ABSTRACT FROM AUTHOR]

Ultraviolet-B (UVB) radiation is a major physical factor that induces structural changes in human skin. The aim of this study was to determine whether the novel silent information regulator 1 (sirtuin 1 SIRT1) protein activator, penilumamide, exerted any protective effects against UVB-induced skin damage using human HaCaT keratinocytes as a model. Enzymatic assays were performed to determine the SIRT1-activating ability of penilumamide, which was compared with that of resveratrol, a potent natural product SIRT1 activator with antioxidant and anti-inflammatory properties. Penilumamide markedly activated SIRT1 enzyme activity compared to resveratrol. To further investigate the protective effect of penilumamide against UVB-induced cytotoxicity, HaCaT cells were pretreated with penilumamide (10 μM) for 24 hr followed by irradiation with UVB (40 mJ/cm2). UVB (40 mJ/cm2) irradiation significantly reduced cell viability in a time-dependent manner, whereas pretreatment with penilumamide blocked this effect. Further, penilumamide decreased the levels of intracellular reactive oxygen species (ROS) generated by UVB irradiation in HaCaT cells. Pretreatment with penilumamide also prevented UVB irradiation-induced changes in mitochondrial membrane potential (ΔΨm). In addition, pretreatment with penilumamide significantly reduced the expression levels of pro-inflammatory cytokines, interleukin (IL)-6, IL-8, and IL-10 and phosphorylation of nuclear factor-kB (NF-kB). These results indicate that penilumamide protects HaCaT cells from UVB-induced inflammation. Taken together data demonstrate that penilumamide exerted protective effects against UVB-induced ROS generation in HaCaT cells. Therefore, penilumamide may be considered to be used as a new SIRT1 activator to protect human keratinocyte against UVB-induced damage. [ABSTRACT FROM AUTHOR]

This study investigated the antibacterial capacity and mechanism of surfactin (prepared by fermenting soybean meal with Bacillus subtilis SOPC5) against Staphylococcus aureus by examining its influences on the bacterial morphology, cell wall and membrane, as well as metabolic activity. The efficiency of surfactin in pork preservation was also investigated. HPLC-MS analysis showed that surfactin's purity reached 97.4%, containing five congeners. The minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) of surfactin against S. aureus were 1.6 and 3.2 mg/mL, respectively. The result of scanning electron microscopy observation revealed that 4 h of surfactin exposure resulted in deformation and even collapse of S. aureus. AKP assay, membrane potential analysis, extracellular protein and nucleic acid content measurement, and PI staining intensity assay showed that surfactin destroyed the cell wall and cell membrane of S. aureus. Decreases in iodonitrotetrazolium chloride levels and ATP enzyme activity indicated that surfactin could inhibit the metabolic ability of S. aureus. Furthermore, surfactin demonstrated an excellent antibacterial action on S. aureus within 5 days of pork storage without changing its texture. These findings suggest that sufactin has a great application potential in pork preservation. [ABSTRACT FROM AUTHOR]

Citrinin (CIT), a mycotoxin commonly found in cereals, is produced by fungi from the Aspergillus, Penicillium, and Monascus genera. While its nephrotoxic effects are well studied, its impact on neurons is less understood. This study investigates CIT-induced toxicity in human neuroblastoma cells (SH-SY5Y). The IC50 values for cells treated with CIT were 77.1 μM at 24 h and 74.7 μM at 48 h using MTT assay, and 101.0 μM at 24 h and 54.7 μM at 48 h using neutral red assay. CIT exposure caused G2/M phase arrest, with cells in this phase increasing from 11.83% (control) to 33.10% at 50 μM CIT. At 50 μM, the percentage of cells in the S phase also increased, which may suggest that cellular stress pathways were activated. Moreover, an increase in late apoptosis process was noted in cells exposed to CIT for 24 h, particularly at the highest concentrations (38.75 and 50 µM). Western blot analysis confirmed a rapid change in the anti-apoptotic protein Bcl-2, but no significant changes in Bax. In conclusion, CIT induces apoptosis and cell cycle arrest in SH-SY5Y cells. However, further transcriptomic studies in specific proteins involved in different pathways described in this work are needed to gain a comprehensive understanding of the specific mechanisms underlying CIT's toxicity in SH-SY5Y cells. [ABSTRACT FROM AUTHOR]

In this study, a chitosan-based polymer composite hydrogel grafted with polyimidazolium was synthesized and evaluated as an adsorbent for nitrate removal from water. The synthesis began with the preparation of a dinitro compound, followed by the synthesis of a diamine monomer, which was then conjugated with chitosan in the presence of glyoxal and formaldehyde to form the final composite. The chemical structure of the synthetic monomers and the composite was characterized using FT-IR, 1H-NMR, FE-SEM, EDX, and TGA analyses. The concentration of nitrate ions in aqueous solutions was quantified using a spectrophotometer at wavelengths of 220 nm and 275 nm. Various operational parameters, including pH, contact time, initial nitrate ion concentration, and adsorbent dosage, were optimized to maximize nitrate removal efficiency. At an initial nitrate concentration of 70 mg/L, a pH of 7, an adsorbent dosage of 40 mg, and a contact time of 40 min, the synthesized composite exhibited a maximum removal efficiency of 97.75% and an adsorption capacity of 85.53 mg/g. The selectivity of the composite for nitrate ions in the presence of competing ions such as sulfate, bicarbonate, chloride, and phosphate was also evaluated. The presence of competing ions reduced nitrate removal, with bicarbonate having the most significant inhibitory effect and phosphate the least. The adsorption kinetics was best described by a Pseudo-second-order model, while the equilibrium data conformed to the Langmuir isotherm model. The PCH reusability was checked by an adsorption/desorption experiment, and the results showed acceptable constant loading efficiency after five reuse cycles. In addition to the adsorption studies, the composite's anti-cancer properties were assessed. Cytotoxicity tests were conducted in vitro against three cancer cell lines: A431 (skin carcinoma), HT29 (colorectal cancer), and MCF7 (breast cancer), using cisplatin as a reference standard. The results demonstrated a potent anti-cancer effect against the MCF7 cell line, with an IC50 value of 4.80 µM. Furthermore, the composite included a collapse in the mitochondrial membrane potential (MMP) in MCF7 cells, highlighting its potential as a therapeutic agent. [ABSTRACT FROM AUTHOR]

Disparate pathogenic mechanisms complicate precision-medicine efforts to treat diffuse large B-cell lymphoma (DLBCL), the most common lymphoma diagnosis. Though potentially curable with frontline combination chemoimmunotherapy, DLBCL carries persistently poor prognosis for those with relapsed or refractory (rel/ref) disease, despite recent advances in immunotherapy. Here, we build on recent findings implicating gain-of-function mutations in the BCL10 signaling protein as drivers of resistance to Bruton's tyrosine kinase (BTK) inhibitors. We show mutant BCL10-driven DLBCL is resistant to multiple additional drug classes, demonstrating urgency to derive mechanistically rooted strategies to overcome undruggable BCL10 mutants that stabilize BTK-independent signaling filaments upstream of NF-kB activation. BCL10 mutants promote a cytokine-reinforced positive feedback loop of lymphomagenesis driving not just NF-kB but multiple additional pathways converging on diffuse activation of oncogenic transcription factors. Up-regulation of anti-apoptotic genes increases mitochondrial membrane potential, underlying multidrug resistance. Increased expression of BCL2, BCL2L1 (BCL-XL), and BCL2A1 (BFL1) drives resistance to venetoclax, but expression can be overcome by the potent non-covalent BTK inhibitor pirtobrutinib. Venetoclax plus pirtobrutinib synergized in overcoming resistance and potently killed BCL10-mutant lymphomas in vitro and in vivo. BTK therefore retains key roles protecting DLBCL from apoptosis even when downstream activation of the BCL10 signaling complex activates NF-kB independently. [ABSTRACT FROM AUTHOR]

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