Membrane Changes Research Articles

Real-time prediction of heparin concentration in blood extracorporeal circulation by relaxation time distribution (RTD).

Heparin concentration c in a blood extracorporeal circulation has been real-timely predicted based on the relaxation strength Δεm at relaxation frequency fm extracted by relaxation time distribution (RTD). The simulated extracorporeal circulation was conducted to optimize the number of Δεm for the prediction of c using the porcine whole blood (WB) and low-leukocyte and -platelet blood (LLPB) under the condition of the gradual increment of c from 0 to 8 U/mL with constant flow rate and blood temperature. The experimental results show that among the three relaxation strengths Δε1, Δε2 and Δε3 (in ascending order of frequency), Δε2 at f2=5.2∼6.2MHz and Δε3 at f3=42∼50MHz were correlated to c. The Δε3 was decreasing with increasing c in both cases, which was influenced by the plasma macromolecular concentrations, while the Δε2 was increased with increasing c in WB case but was hardly changed in LLPB case because the Δε2 is influenced by the blood cell concentrations and the shape changes of blood cell membranes. Heparin concentration c is estimated by the linear regression formula cPRE=a1(Δε2-Δε2c=0)+a2(Δε3-Δε3c=0) (a1=-0.991, a2=-0.123) within the mean absolute percentage error (MAPE) of 0.291.

Heterogeneity in Fluorescence-Stained Sperm Membrane Patterns and Their Dynamic Changes Towards Fertilization in Mice.

Sperm represent a heterogeneous population crucial for male reproductive success. Additionally, sperm undergo dynamic changes during maturation and capacitation. Despite these well-established processes, the complex nature of sperm heterogeneity and membrane dynamics remains elusive. The composition of phospholipids in the sperm membrane changes dynamically during maturation, with their release occurring during capacitation. This study aims to investigate the heterogeneity and dynamic changes in the sperm membrane during maturation and capacitation towards fertilization by visualizing these membrane dynamics. Sperm were collected from the cauda epididymis or testis of Institute of Cancer Research (ICR) male mice and stained with MemBright dye (commercial name: MemGlow™-560, MG-560), a fluorogenic live-cell membrane probe. Staining was performed either before, during, or after incubation for capacitation. In pre-staining experiments, sperm were stained with MG-560 before capacitation and then incubated to induce capacitation. Acrosome-reacted sperm were assessed after staining with peanut agglutinin FITC (PNA-Lectin FITC). Stained sperm were observed using fluorescence or confocal microscopy. MG-560-stained sperm from the epididymis before capacitation showed four staining patterns: head-midpiece-tail (HMT), head-midpiece (HM), head (H), midpiece (M) positive, or totally negative, with ratios remaining unchanged during capacitation (30.5%, 29%, 11.3%, 3.7%, and 25.5%, respectively). In contrast, all testicular sperm were negative for staining. Pre-stained sperm exhibited an increased number of HM and M patterns over time, whereas the number of HMT-stained sperm decreased. Consistently, spontaneous acrosome-reacted sperm were detected predominantly in HM- or M-stained sperm. After in vitro fertilization (IVF) using pre-stained sperm, zona pellucida-attached sperm were mostly negative for staining. Finally, all sperm detected in the perivitelline space were only negative. Mature sperm membranes stained with MG-560 exhibited heterogeneous and dynamic changes during the capacitation and fertilization process. MG-560 staining identified sperm with the potential to undergo the acrosome reaction, and these MG-560-positive sperm eventually became negative as they penetrated the zona pellucida for fertilization. Thus, the MG-560 staining patterns likely reflect the physiological state and potential of the sperm. These findings provide new insights into sperm heterogeneity and dynamics, and this staining method may also prove useful for assessing sperm quality.

Plant Coping with Cold Stress: Molecular and Physiological Adaptive Mechanisms with Future Perspectives.

Cold stress strongly hinders plant growth and development. However, the molecular and physiological adaptive mechanisms of cold stress tolerance in plants are not well understood. Plants adopt several morpho-physiological changes to withstand cold stress. Plants have evolved various strategies to cope with cold stress. These strategies included changes in cellular membranes and chloroplast structure, regulating cold signals related to phytohormones and plant growth regulators (ABA, JA, GA, IAA, SA, BR, ET, CTK, and MET), reactive oxygen species (ROS), protein kinases, and inorganic ions. This review summarizes the mechanisms of how plants respond to cold stress, covering four main signal transduction pathways, including the abscisic acid (ABA) signal transduction pathway, Ca2+ signal transduction pathway, ROS signal transduction pathway, and mitogen-activated protein kinase (MAPK/MPK) cascade pathway. Some transcription factors, such as AP2/ERF, MYB, WRKY, NAC, and bZIP, not only act as calmodulin-binding proteins during cold perception but can also play important roles in the downstream chilling-signaling pathway. This review also highlights the analysis of those transcription factors such as bHLH, especially bHLH-type transcription factors ICE, and discusses their functions as phytohormone-responsive elements binding proteins in the promoter region under cold stress. In addition, a theoretical framework outlining plant responses to cold stress tolerance has been proposed. This theory aims to guide future research directions and inform agricultural production practices, ultimately enhancing crop resilience to cold stress.

Rapid neurostimulation at the micron scale with an optically controlled thermal-capture technique.

Precise control of cellular temperature at the microscale is crucial for developing novel neurostimulation techniques. Here, the effect of local heat on the electrophysiological properties of primary neuronal cultures and HEK293 cells at the subcellular level using a cutting-edge micrometer-scale thermal probe, the diamond heater-thermometer (DHT), is studied. A new mode of local heat action on a living cell, thermal-capture mode (TCM), is discovered using the DHT probe. In TCM, the application of a 50 °C temperature step induces a great increase in cellular response, allowing the cell to be thermally captured and depolarized by up to 20 mV. This thermal effect is attributed to local phase changes in the phospholipid membrane, enabling precise and reproducible modulation of cell activity. The TCM is shown to open up new opportunities for thermal cell stimulation. DHT reliably triggers action potentials (APs) in neurons at rates up to 30 Hz, demonstrating the ability to control cell excitability with millisecond and sub-millisecond resolution. AP shape is modulated by local heat as well. The ability to precisely control the AP shape and rate via thermal-capture mode opens new avenues for non-invasive, localized neurostimulation techniques, particularly in controlling neuron excitability.

Corticostriatal maldevelopment in the R6/2 mouse model of juvenile Huntington's disease.

There is a growing consensus that brain development in Huntington's disease (HD) is abnormal, leading to the idea that HD is not only a neurodegenerative but also a neurodevelopmental disorder. Indeed, structural and functional abnormalities have been observed during brain development in both humans and animal models of HD. However, a concurrent study of cortical and striatal development in a genetic model of HD is still lacking. Here we report significant alterations of corticostriatal development in the R6/2 mouse model of juvenile HD. We examined wildtype (WT) and R6/2 mice at postnatal (P) days 7, 14, and 21. Morphological examination demonstrated early structural and cellular alterations reminiscent of malformations of cortical development, and ex vivo electrophysiological recordings of cortical pyramidal neurons (CPNs) demonstrated significant age- and genotype-dependent changes of intrinsic membrane and synaptic properties. In general, R6/2 CPNs had reduced cell membrane capacitance and increased input resistance (P7 and P14), along with reduced frequency of spontaneous excitatory and inhibitory synaptic events during early development (P7), suggesting delayed cortical maturation. This was confirmed by increased occurrence of GABAA receptor-mediated giant depolarizing potentials at P7. At P14, the rheobase of CPNs was significantly reduced, along with increased excitability. Altered membrane and synaptic properties of R6/2 CPNs recovered progressively, and by P21 they were similar to WT CPNs. In striatal medium-sized spiny neurons (MSNs), a different picture emerged. Intrinsic membrane properties were relatively normal throughout development, except for a transient increase in membrane capacitance at P14. The first alterations in MSNs synaptic activity were observed at P14 and consisted of significant deficits in GABAergic inputs, however, these also were normalized by P21. In contrast, excitatory inputs began to decrease at this age. We conclude that the developing HD brain is capable of compensating for early developmental abnormalities and that cortical alterations precede and are a main contributor of striatal changes. Addressing cortical maldevelopment could help prevent or delay disease manifestations.

Biopreservation and reversal of oxidative injury during blood storage by a novel curcumin-based gel formulation

Blood storage lesion induces cytosolic and membrane changes driven in part by hemoglobin (Hb) oxidation reactions within red blood cells (RBCs). A novel gel formulation containing the antioxidant curcuminoids in a biocompatible solvent system was used to deliver curcumin into RBCs. Incubation of peroxide treated RBCs stored in PBS with curcumin gel led to a reduction in prooxidant ferrylHb and recovery in ATP. Curcumin treatment prevented band 3 tyrosine (Y359 and Y21) phosphorylation. RBCs stored in AS-3 solutions for 28, 35, 42 and 49 days, following a single-dose of 100µM curcuminoids at each time points, caused reduction in protein carbonylation and considerable recovery in ATP levels. Proteomic analysis revealed minimal changes in the proteomic landscape in 35 days. However, a downregulation in fibrinogen was observed in the treated samples which may reduce RBC aggregation. Additionally, we used a guinea pig model where the circulation of infused aged RBCs can be extended (approximately 10%) when treated with curcumin gel at the start of storage. Our data therefore provide mechanistic insights and supportive animal data into benefits of treating stored RBCs with a novel curcuminoid formulation based on the biopreservation of RBC membrane integrity, redox balance, and increased longevity in circulation.

Melittin-Induced Structural Transformations in DMPG and DMPS Lipid Membranes: A Langmuir Monolayer and AFM Study.

In this study, we explore the interactions between melittin, a cationic antimicrobial peptide, and model lipid membranes composed of the negatively charged phospholipids 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) and 1,2-dimyristoyl-sn-glycero-3-phosphoserine (DMPS). Using the Langmuir monolayer technique and atomic force microscopy (AFM), we reveal novel insights into these interactions. Our key finding is the observation of the ripple phase in the DMPS bilayer on mica, a phenomenon not previously reported for negatively charged single bilayers. This discovery is significant given the critical role of phosphatidylserine (PS) in cancer biology and the potential of melittin as an anticancer agent. We also highlight the importance of subphase composition, as melittin interacts preferentially with lipids in the liquid-condensed phase; thus, selecting the appropriate subphase composition is crucial because it affects lipid behavior and consequently melittin interactions. Our results show that melittin incorporates into lipid monolayers in both liquid-expanded and liquid-condensed phases, enhancing membrane fluidity and disorder, but is expelled from DMPS in the solid phase. AFM imaging further reveals that melittin induces substantial structural changes in the DMPG membrane and forms the ripple phase in the DMPS bilayers. Despite these alterations, melittin does not cause pore formation or membrane rupture, suggesting strong electrostatic adsorption on the membrane surface that prevents penetration. These findings highlight the differential impacts of melittin on lipid monolayers and bilayers and underscore its potential for interacting with membranes without causing disruption.

Understanding Bacterial Resistance to Heavy Metals and Nanoparticles: Mechanisms, Implications, and Challenges.

Antimicrobial resistance is a global health problem as it contributes to high mortality rates in several infectious diseases. To address this issue, engineered nanoparticles/nano-formulations of antibiotics have emerged as a promising strategy. Nanoparticles are typically defined as materials with dimensions up to 100nm and are made of different materials such as inorganic particles, lipids, polymers, etc. They are widely dispersed in the environment through various consumer products, and their clinical applications are diverse, ranging from contrast agents in imaging to carriers for gene and drug delivery. Nanoparticles can also act as antimicrobial agents either on their own or in combination with traditional antibiotics to produce synergistic effects, earning them the label of "next-generation therapeutics." They have also shown great effectiveness against multidrug-resistant pathogens responsible for nosocomial infections. However, overexposure or prolonged exposure to sublethal doses of nanoparticles can promote the development of resistance in human pathogens. The resistance can arise from various factors such as genetic mutation, horizontal gene transfer, production of reactive oxygen species, changes in the outer membrane of bacteria, efflux-induced resistance, cross-resistance from intrinsic antibiotic resistance determinants, plasmid-mediated resistance, and many more. Continuous exposure to nanoparticles can also transform an antibiotic-susceptible bacterial pathogen into multidrug resistance. Considering all these, the current review focuses on the mode of action of different heavy metals and nanoparticles and possible mechanisms through which bacteria attain resistance towards these heavy metals and nanoparticles.

ARGONOPLASMIC COAGULATION IN THE TREATMENT OF CHRONIC RADIATION PROCTITIS COMPLICATED BY BLEEDING

Rationale: Chronic radiation proctitis is a complication of remote radiation therapy used to treat malignant neoplasms of the pelvis, and is often complicated by the development of rectal bleeding. Argonoplasmic coagulation is one of the available endoscopic methods of their treatment, but the effectiveness of its use needs to be studied.Objective: To evaluate the effectiveness of endoscopic argonoplasmic coagulation to eliminate telangiectasias of the rectal mucosa in patients with chronic radiation proctitis complicated by bleeding, and to present a sectoral classification of lesions of the rectal mucosa with telangiectasias.Methods: The article presents the results of treatment of 18 patients with chronic radiation proctitis who underwent argonoplasmic coagulation of telangiectasias of the mucous membrane of the rectum in order to stop bleeding and prevent their occurrence. To assess the effectiveness of treatment, a sectoral classification of the rectal lesion was applied.Results: After argonoplasmic coagulation of telangiectasia of the rectal mucosa, all patients had no rectal bleeding for 6 months. In 13 (72%) patients, there was a sharp decrease in the frequency of blood excretion during defecation, in the remaining 5 (28%) patients there was no blood excretion from the rectum. In the postoperative period, two recurrence of bleeding from the treatment site was observed on the second and seventh days of the postoperative period, which required repeated application of argonoplasmic coagulation.Conclusion: Endoscopic argonoplasmic coagulation is effective in eliminating telangiectasia in patients with chronic radiation proctitis complicated by bleeding. The presented sectoral classification of rectal lesions in chronic radiation proctitis is convenient to use and contributes to the assessment of changes in the mucous membrane of the rectum in dynamics.

Navigating the complex role of senescence in liver disease.

Cellular senescence, an irreversible state of cell cycle arrest characterized by phenotypic changes and a specific secretory profile, plays a dual role in liver health and disease. Under physiological conditions, senescence aids organ repair and regeneration, but its accumulation due to aging or pathological stress significantly contributes to chronic liver diseases, including alcoholic liver disease, metabolic dysfunction-associated steatohepatitis, liver fibrosis, and hepatocellular carcinoma. Senescence is identified by a range of cellular and molecular changes, such as morphological alterations, expression of cell cycle inhibitors, senescence-associated β-galactosidase activity, and nuclear membrane changes. The onset of senescence in organ cells can affect the entire organism, primarily through the senescence-associated secretory phenotype, which has autocrine, paracrine, and endocrine effects on tissue microenvironments. The objective of this review is to offer a contemporary overview of the pathophysiological events involving hepatic senescent cells and to elucidate their role in the onset and progression of liver diseases, particularly through mechanisms like telomere shortening, genomic and mitochondrial DNA damage, and inflammation. Additionally, this review discusses the emerging senolytic therapies aimed at targeting senescent cells to delay or mitigate liver disease progression. The therapeutic potential of these interventions, alongside their safety and effectiveness, highlights the need for further research to refine these approaches and address unresolved problems in the field of hepatic cellular senescence.

Consistent use injectable and oral chondroprotectors in patients with osteoarthritis and lower back pain. Case report

Osteoarthritis (OA) is a disease of the entire joint, including structural changes in hyaline articular cartilage, subchondral bone, ligaments, capsule, synovial membrane and periarticular muscles. One of the causes of the development of nonspecific chronic lower back pain (LBP) is OA of the facet joints. In the treatment of OA and LBP against the background of OA facet joints, the chondroprotective agents are widely used: chondroitin sulfate (CS), glucosamine sulfate, undenatured type II collagen and their combinations. The aim of the article is to systematize the possibilities of practical application of Chondroguard TRIO® in patients with OA of different phenotypes and different localization. The experience of sequential administration of CS in patients (n=11) with OA and LBP with comorbid diseases is presented. At the initial stage of therapy, CS (Chondroguard®) was prescribed intramuscularly according to the scheme, then CS was prescribed perorally as part of a pharmaconutraceutical (Chondroguard® TRIO). The original pharmaconutraceutical Chondroguard® TRIO contains recommended doses of CS (1200 mg), glucosamine sulfate (1500 mg), undenatured type II collagen (40 mg). Clinical cases of the use of sequential administration of chondroprotectors (stage 1 – intramuscularly Chondroguard®, stage 2 – perorally Chondroguard® TRIO) demonstrate the clinical effectiveness of the proposed treatment regimen in patients of different age groups and OA phenotypes (posttraumatic, metabolic).

Microcirculatory changes in the mucous membrane of the cervix in women with isthmic-cervical insufficiency

Microcirculatory changes in the mucous membrane of the cervix in women with isthmic-cervical insufficiency

Optical Imaging of Trigeminal Ganglion Excitation Evoked by Electrical Stimulation of the Trigeminal Nerve.

Background There are many reports of anatomical and physiological studies on trigeminal ganglion neurons, but few studies have analyzed temporal changes in the excitation of the trigeminal ganglion. This study aimed to establish an experimental system for spatial and temporal imaging analysis of the excitatory dynamics of trigeminal ganglion cells evoked by stimulation of a peripheral branch of the trigeminal nerve. Methods After excision of the trigeminal ganglion with the inferior alveolar nerve (IAN) from Sprague Dawley rats (seven to nine weeks old), 400-µm-thick slices of the trigeminal ganglion with the IAN were prepared. Real-time optical imaging was performed using cell membrane voltage-sensitive dye, Di4-ANEPPS, and changes in fluorescence intensity ratio (ΔF/F) were analyzed. Results Electrical stimulation of the IAN evoked the excitation of the trigeminal ganglion at the lateral surface area first, followed by expansion to the inner area. The calculated conduction velocity was 0.72 ± 0.49 m/s. This response was diminished by tetrodotoxin perfusion, but it was not observed in the C fiber-deficient rats. Conclusions A real-time optical imaging system can visualize the excitation of the trigeminal ganglion evoked by C-fiber stimulation.

Refractive Index Morphology Imaging Microscope System Utilizing Polarization Multiplexing for Label-Free Single Living Cells.

Detections of internal substances and morphologies for label-free living cells are crucial for revealing malignant diseases. With the phase serving as a coupling of refractive index (RI) (marker for substances) and thickness (morphology), existing decoupling methods mainly rely on complex integrated systems or extensive optical field information. Developing simple and rapid decoupling methods remains a challenge. This study introduces a refractive index morphology imaging microscope (RIMIM) system utilizing polarization multiplexing for label-free single living cells. By simultaneous degree of circular polarization (DOCP) imaging and noninterferometric quantitative phase imaging (QPI), the intracellular refractive index distribution (IRID) and morphology can be decoupled. The optical thickness calculated from the phase is input into the circular depolarization decay model (CDDM) of degree of circular polarization to retrieve IRID. Subsequently, the thickness can be decoupled from phase result using retrieved IRID. Experiments conducted on mouse forestomach carcinoma (MFC) cells and human kidney-2 cells (HK-2) demonstrated the RIMIM system's ability to retrieve IRID and decouple fine morphology. Additionally, the RIMIM system effectively detected membrane damage and changes in erastin-induced ferroptotic HK-2 cells, with average and root-mean-square of surface folds 65.5% and 70.0% higher than those of normal HK-2 cells. Overall, the RIMIM system provides a simple and rapid method for decoupling RI and fine morphology, showing great potential for label-free live cells' cytopathology detection.

Diagnosis and Treatment of Gastroesophageal Reflux Disease (Clinical Guidelines of the Russian Gastroenterological Association, Russian Scientific Medical Society of Internal Medicine, Russian Society for the Prevention of Noncommunicable Diseases, Scientific Community for Human Microbiome Research)

Aim. These recommendations are developed for practitioners in order to familiarize them with modern diagnostic methods, management features and pharmacotherapy of patients with gastroesophageal reflux disease (GERD).General provisions. GERD is the most common reason for patients to visit clinics. There are esophageal and extraesophageal manifestations of GERD. Patients' complaints of heartburn and regurgitation remain the most sensitive and specific clinical manifestations of GERD. The diagnosis of GERD is established on the basis of anamnestic data, instrumental examination (detection of reflux esophagitis during upper gastrointestinal endoscopy, detection of pathological gastroesophageal reflux with 24-hour pH-metry or/and 24-hour pH-impedance monitoring). Patients with suspected GERD and the absence of erosive and ulcerative changes in the mucous membrane of the esophagus or the presence of erosive esophagitis of Grade A according to Los Angeles Classification of Gastroesophageal Reflux Disease are recommended to conduct 24-hour pH-metry on PPI off to exclude or confirm the diagnosis of GERD. Patients with extraesophageal manifestations of GERD without classic symptoms (heartburn, regurgitation) are recommended to undergo 24-hour pH-impedance monitoring with discontinuation of proton pump inhibitor therapy. When deciding on surgical treatment, all patients need to perform high-resolution esophageal manometry and 24-hour pH-impedance monitoring. Complications of GERD include bleeding, strictures, Barrett’s esophagus and esophageal adenocarcinoma. The main groups of medications used in the treatment of GERD are proton pump inhibitors (PPIs), potassium-competitive acid blockers (P-CABs), alginates, antacids, and prokinetics. PPIs are the drugs of choice in the treatment of both symptoms of gastroesophageal reflux disease and existing erosive esophagitis. Combination therapy Rebamipide with PPIs increases the effectiveness of relief of GERD symptoms, as well as reduces the frequency of relapses.Conclusion. These clinical recommendations will improve the quality of medical care for patients with GERD.

Microrheometric study of damage and rupture of capsules in simple shear flow.

Capsules, which are potentially-active fluid droplets enclosed in a thin elastic membrane, experience large deformations when placed in suspension. The induced fluid-structure interaction stresses can potentially lead to rupture of the capsule membrane. While numerous experimental studies have focused on the rheological behavior of capsules until rupture, there remains a gap in understanding the evolution of their mechanical properties and the underlying mechanisms of damage and breakup under flow. We here investigate the damage and rupture of bioartificial microcapsules made of ovalbumin reticulated with terephthaloyl chloride and placed in simple shear flow. We characterize damage by identifying how the surface shear modulus of the capsule membrane changes over time. Rupture is then characterized by comparing the number and size distribution of capsules before and after exposure to shear, while varying the shear rates and time during which capsules are sheared. Our findings reveal how the percentage of ruptured capsules increases with their size, exposure time to shear, and the ratio of viscous to elastic forces at rupture.

Proteomics and surface free fatty acid analysis of milk fat globule in the spray- and freeze-dried bovine, goat and horse milk powders.

Changes in the structure and composition of milk fat globules in spray- and freeze-dried milk powders have recently garnered significant attention. This study investigated changes in milk fat globular membrane (MFGM) proteins from bovine, goat, and horse milk powders, both spray- and freeze-dried, using a label-free proteomics approach, and quantified surface free fatty acids and their composition using gas chromatography. The results showed that several proteins of αS2-casein and β-lactoglobulin increased, while fibrinogen α, β chain, and mucin-1 decreased in the MFGM fractions of the studied spray-dried milk powders. Additionally, lactoperoxidase and polymeric immunoglobulin receptor levels were elevated in the studied freeze-dried milk powders. Several proteins exhibited variations in both dried milk powders depending on the species, of these, nucleobindin-1, complement C3, and sulfhydryl oxidase were increased in spray-dried bovine and goat milk powders, and lactoferrin was increased in freeze-dried horse milk powder, compared with their raw milk counterparts. Conversely, butyrophilin subfamily 1 member A1 and xanthine dehydrogenase/oxidase were decreased in spray-dried bovine and goat milk powders, S100 calcium-binding protein and aldehyde dehydrogenase were decreased in freeze-dried bovine and goat milk powders, and mucin-4 and paraoxonase were decreased in horse milk powder. Additionally, spray-dried milk powders had lower surface free fatty acid content compared with freeze-dried milk powders. The findings underscore that dried methods exert varied impacts on MFGM components of the studied milk sources, thereby providing a valuable reference for improving the nutritional quality of dried dairy products.

From Germination to Stagnation: Sodium Diacetate's Impact on Clostridium perfringens Spores

ABSTRACTSodium diacetate is recognized for its high efficiency as a preservative, demonstrating strong antibacterial properties that help extend the shelf life of food products. However, it is still unclear how it influences spores. This research investigated the influence of sodium diacetate on the germination and outgrowth stage in the revival process of Clostridium perfringens (C. perfringens) spores. The germination and outgrowth rate of spores after sodium diacetate treatment were measured. The changes of spore inner membrane were monitored by laser confocal microscope, electron scanning microscope and macromolecular leakage. The interaction between sodium diacetate and DNA was investigated by EB fluorescence probe, and the finding was verified by the outgrowth of spores in ham sausage. An investigation into the effects of sodium diacetate treatment on the biological activity, inner membrane permeability, morphology, and DNA integrity of spores devoid of both the coat and cortex revealed that sodium diacetate significantly hindered the transition of germinated spores into vegetative cells. In the presence of sodium diacetate, C. perfringens spores underwent germination, characterized by the hydrolysis of the spore cortex and the disassembly of the spore coat. As the spores advanced into the outgrowth phase, sodium diacetate penetrated their structure, causing damage to the inner membrane and compromising DNA integrity. Moreover, sodium diacetate was demonstrated to effectively inhibit spore outgrowth in ham sausage. This study provided theoretical guidance and references for the application of sodium diacetate in food to control the germination and outgrowth of spore‐forming bacteria.

FUT8 upregulates CD36 and its core fucosylation to accelerate pericyte-myofibroblast transition through the mitochondrial-dependent apoptosis pathway during AKI-CKD

BackgroundActivation of pericytes leads to renal interstitial fibrosis, but the regulatory mechanism of pericytes in the progression from AKI to CKD remains poorly understood. CD36 activation plays a role in the progression of CKD. However, the significance of CD36 during AKI-CKD, especially in pericyte, remains to be fully defined.MethodsGEO and DISCO database were used to analyze the expression of CD36 in pericyte during AKI-CKD; IRI to conduct AKI-CKD mouse model; Hypoxia/Reoxygenation (H/R) to induce the cell model; RT-qPCR and Western blotting to detect gene expression; IP and confocal-IF to determine the core fucosylation (CF) level of CD36. Flow cytometry (AV/PI staining) to detect the cell apoptosis and JC-1 staining to react to the change of mitochondrial membrane potential.ResultsDuring AKI to CKD progression, CD36 expression in pericytes is higher and may be influenced by CF. Moreover, we confirmed the positive association of CD36 expression with pericyte-myofibroblast transition and the progression of AKI-CKD in an IRI mouse model and hypoxia/reoxygenation (H/R) pericytes. Notably, we discovered that FUT8 upregulates both CD36 expression and its CF level, contributing to the activation of the mitochondrial-dependent apoptosis signaling pathway in pericytes, ultimately leading to the progression of AKI-CKD.ConclusionThese results further identify FUT8 and CD36 as potential targets for the treatment in the progression of AKI-CKD.

Photoinactivation mechanism of hypericin nanoencapsulated in P123 against Microsporum canis.

Aim: To evaluate the photodynamic mechanism of hypericin nanoencapsulated in P123 copolymer micelles against Microsporum canis in vitro.Material & methods: Antifungal susceptibility tests were performed, including the determination of the minimum fungicidal concentration and time-kill curve. Flow cytometry was used to evaluate the internalization of P123-Hyp in conidia and the activation of PDT type I and II mechanisms via the detection of reactive oxygen species (ROS), as well as to assess changes in the cell membrane using propidium iodide (PI) and cell morphology.Results: P123-Hyp-PDT exerted a fungistatic action on fungus, maintaining this action up to 24h after exposure, corroborating the PS internalization results, which showed satisfactory uptake of P123-Hyp from a concentration of 3.125μmol/l. Among the ROS studied, singlet oxygen was detected. Furthermore, the increased fluorescence intensity of PI in treated cells indicated necrotic cell death, while the size and granularity of the cells were also altered.Conclusion: Our results show, for the first time, a proposal for the mechanism of action of P123-Hyp-mediated PDT against M. canis, proving that it has a prolonged action on the fungus through activation of the type II photodynamic pathway, which resulted in disruption of the plasma membrane and cellular alterations.