Hemolysis Of Erythrocytes Research Articles

Two‐Phase Extraction, Structural Characterization of the Polysaccharide from Raspberry Pomace and Its Protection Effects against Erythrocyte Hemolysis

AbstractIn present study, a new polysaccharide (RPP) from raspberry pomace is prepared through ultrasound‐assisted aqueous two‐phase extraction (UAATPE) and purification using two‐step column chromatography, the extraction conditions are optimized by response surface method (RSM). Under the optimal parameters: liquid to material ratio of 80 mL g−1, ultrasonic power of 340 W, and ultrasonic time of 39 min, the yield of the polysaccharides is 11.72 ± 0.05%. RPP with average molecular weight (Mw) of 59 300 Da is composed of six monosaccharides. Nuclear magnetic resonance (NMR) analyses confirmed that RPP contains six main glycosidic linkages including α‐l‐Araf‐(1→, →2)‐α‐l‐Rhap‐(1→, →4)‐β‐d‐Manp‐(1→, →4)‐α‐d‐GalAp‐(1→, →3)‐β‐d‐Galp‐(1→, →4)‐β‐d‐Glcp‐(1→. RPP exhibits remarkable protective effects against H2O2‐induced erythrocyte hemolysis, the generations of reactive oxygen species, and malondialdehyde are inhibited, and the activities of glutathione peroxidase and catalase are effectively enhanced. The current findings proved that UAATPE is an effective method to extract the polysaccharide from raspberry pomace and provides a new idea for the utilization of nutrient‐rich raspberry fruit residue.

Regulation of Glass Transition Temperature in Thermo‐Responsive Epoxy Shape Memory Polymers: The Roles of Chemical Crosslinking Density and Chain Flexibility

ABSTRACTShape memory polymers (SMPs) are novel kinds of smart materials whose shape can be changed in external conditions such as heat stimulus. Due to their excellent structural versatility and high elastic strain, they have important applications in the field of biomaterials. Here, a series of amorphous shape memory polymer materials are prepared and their shape memory properties and glass transition temperature (Tg) are also investigated. A special kind of epoxy polymer, whose shape can be changed at around physiological temperature, is synthesized by regulating the crosslinking density and introducing flexible aliphatic epoxy chains. The shape memory function of this epoxy polymer is studied by deformation test. In addition, to explore their application in the field of biomedical materials, the biocompatibility of the materials are evaluated, including the effects on the erythrocyte hemolysis rate and cell activity. The experimental results show that this kind of epoxy polymer has good shape memory function and biocompatibility.

Design, Synthesis, and Biological Studies of C-5-Substituted Diazenyl Derivatives of Uracil as Potent and Selective Antileishmanial Agents Targeting Uridine Biosynthesis Pathway Enzymes.

Herein, we describe the design and synthesis of a series of C-5-substituted diazenyl derivatives of uracil, exhibiting selective and potent antileishmanial but not antibacterial or antifungal activity. The formation of the substituted derivatives was confirmed by using FTIR, 1H, 13C NMR, and HRMS analysis. Among all of the sets of tested compounds, only three [4a, 6b, and 8b] showed the highest activity against Leishmania donovani (LD) promastigote and amastigote models of LD infections. Further, the cytotoxicity assays performed using three different cell lines, Vero cells, J774 cells, and THP1 cells, along with erythrocyte hemolysis assay showed the highest biocompatibility for the 4a, making it a lead compound for further biological assays. The LD cell death associated with 4a was not linked with ergosterol depletion, a common mechanism of action of antileishmanial drugs like amphotericin B (AmB). However, the LD cell death in the presence of 4a was reversed significantly through supplementation of uridine monophosphate (UMP), indicating the specific role of uridine biosynthesis pathway as the target of 4a. Furthermore, the in silico studies predicted ornithine monophosphate decarboxylase enzyme (OMPDCase) from LD as the plausible target for 4a. The proteomics analysis showed stronger downregulation of the aforementioned OMPDCase and also for a few other enzymes that are involved in the UMP biosynthesis pathway. This indicates that OMPDCase and other enzymes that regulate the UMP biosynthesis may be the target of 4a. Overall, the C-5-substituted diazenyl derivatives of uracil are presented here as novel and potent antileishmanial agents that can be used for treating visceral leishmaniasis (VL) wherein at present drug resistance and side effects of existing drugs demand a look for safer alternatives.

Aframomum Melegueta: Evaluation of Chronic Toxicity, HPLC Profiling, and In Vitro/In Vivo Antioxidant Assessment of Seeds Extracts.

Aframomum melegueta, commonly known as grains of paradise, is a medicinal plant celebrated for its rich phytochemical content and therapeutic properties. This study evaluated the antioxidant and cytotoxic potentials of its ethanolic and methanolic extracts, both in vitro and in vivo, while also analyzing their chemical profiles. HPLC analysis identified key compounds, including gallic acid, caffeic acid, caffeine, coumarin, rutin, catechin, ferulic acid, and quercetin. Chronic toxicity assessments confirmed the safety of the extracts, with no adverse effects on animal health, particularly in liver histopathology. Cytotoxicity results indicated reduced splenocyte viability at the highest concentrations. The extracts exhibited significant antioxidant activity in DPPH•, FRP, and phosphomolybdate assays, demonstrating their effectiveness as antiradical agents. In vivo antioxidant results showed a reduction in lipid peroxidation levels in serum and liver, highlighting the extracts' ability to mitigate oxidative stress. Additionally, the extracts provided protection against H2O2-induced erythrocyte hemolysis and modulated NO production in peritoneal macrophages. These findings underscore the therapeutic potential of A. melegueta extracts, suggesting their promise in developing preventive strategies for oxidative stress-related chronic diseases.

In Vitro Antitumor, Antioxidant, and Hemolytic Activities of Chlorella sorokiniana Methanol Extracts and Collective Fractions

Chlorella species are fast-growing microalgae with significant industrial applications. The aim of the present study was to investigate the antitumor, antioxidant, and hemolytic activities of Chlorella sorokiniana UTEX 1230 crude methanol extracts and fractions. Ch. sorokiniana crude methanol extracts and collective fractions (CFs) were obtained from lyophilized biomass by maceration and column chromatography. Antitumor assays against murine lymphoma L5178Y-R and human breast cancer MCF-7 cells were performed by the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction technique, using human peripheral blood mononuclear cells (PBMC) as the control group. Antioxidant and hemolytic activities were evaluated using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay (DPPH) and erythrocyte hemolysis, respectively. We showed that crude methanol extracts (IC50) increased L5178Y-R and MCF-7 cell growth inhibition, without affecting PBMC. In addition, all evaluated CFs showed significantly higher antioxidant activity than the positive control (ascorbic acid). CF3 and CF4 showed the highest cytotoxicity against L5178Y-R, whereas CF3, CF4, and CF5 caused the highest antitumor activity against MCF-7 cells. CF3, CF4, and CF5 induced significantly higher hemolytic activity compared with all other fractions. CF characterization revealed loliolide, cinnamic acid, methyl dihydrojasmonate, salsalvamide A, 1-monolinolenin, cryptophycin 29, costunolide, riboflavin lumicrome, and germicidin B, which have been related to antitumor and antioxidant activities. In conclusion, we demonstrated that Ch. sorokiniana extracts and fractions possess antitumor and antioxidant potential, without affecting human erythrocytes and PBMC.

Levofloxacin induces erythrocyte contraction leading to red cell death.

Levofloxacin, a fluoroquinolone, is an extensively used antibiotic effective against both positively and negatively staining bacteria. It works by inhibiting bacterial topoisomerase type II and topoisomerase type IV, resulting in impaired DNA synthesis and bacterial cell death. Eryptosis is another term for apoptotic cell death of erythrocyte marked by cell shrinkage, phosphatidylserine (PS) flipping, and membrane blebbing. The intent of the present research was to look at the eryptotic effect of levofloxacin by exposing erythrocytes to therapeutical doses (7, 14 µM) of levofloxacin for 48 hours. Cell size evaluation, PS subjection to outside, and calcium channel inhibition were carried out to investigate eryptosis. Oxidative stress generated by levofloxacin was measured as a putative mechanism of eryptosis using glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase activities. Similarly, hemolysis measurements demonstrated levofloxacin's cytotoxic effect. Our findings showed that therapeutic doses of levofloxacin can cause a considerable decline in antioxidant enzymes activities, as well as induce cell shrinkage, PS externalization, and hemolysis in erythrocytes. The role of calcium in triggering erythrocyte shrinkage was also confirmed. In conclusion, our findings showed that the indicated levofloxacin doses caused oxidative stress, which leads to erythrocyte death via eryptosis and hemolysis. These findings emphasize the importance of using levofloxacin with caution and the need for additional research to mitigate these side effects.

In Vitro Hemocompatibility of Arabinogalactan, Betulin, and Betulin Derivatives.

The hemocompatibility of arabinogalactan, betulin and its derivatives was evaluated in vitro and samples suitable for creation of nanostructures or materials in contact with blood were selected. The prospects of arabinogalactan as a component of the construct (nanostructure) for drug delivery are due to the fact that it did not affect blood/plasma coagulation (at concentrations of 0.0033-3.333 mg/ml and 0.00465-4.65 mg/ml, respectively), platelet aggregation (0.00182-0.182 mg/ml), and demonstrated the degree of erythrocyte hemolysis less than 3%. Sodium salt of betulin monosulfate, diarginine salt of betulin disulfate (up to 0.465 mg/ml), and especially betulin and allobetulin formate with procoagulant properties (degree of hemolysis less than 2%) can be used to create a material, for example, sponge, gel, active against blood coagulation.

Glycyrrhetinic acid reduces lung inflammation caused by pneumococcal infection by reducing the toxicity of pneumolysin

ObjectiveIn this study, to provide new methods for the treatment of Streptococcus pneumoniae infection, we aimed to describe the anti-inflammatory and antibacterial value of glycyrrhetinic acid on the basis of its inhibitory effect on bacterial growth (without killing the bacteria) and its reduction of the toxicity of S. pneumoniae. MethodsA mouse model was established via intranasal administration of Streptococcus pneumoniae D39, and glycyrrhetinic acid was subcutaneously injected for treatment. The wet‒dry ratio, bacterial flora content and inflammatory factor levels in the mouse lungs were determined. Cell experiments were used to evaluate glycyrrhetinic acid-mediated inhibition of PLY hemolysis and A549 cell death, and WB was used to measure glycyrrhetinic acid-mediated inhibition of PLY oligomerization. ResultsGlycyrrhetinic acid reduced the levels of inflammatory factors, the dry‒wet ratio, the abundance of S. pneumoniae in the lungs of infected mice, pneumolysin-mediated A549 cell death, erythrocyte hemolysis and PLY oligoplasia. ConclusionGlycyrrhetinic acid can reduce the virulence of S. pneumoniae by preventing the oligomerization of PLY.

Antimicrobial spectrum against wound pathogens and cytotoxicity of star-arranged poly-l-lysine-based antimicrobial peptide polymers.

Introduction. As growing numbers of patients are at higher risk of infection, novel topical broad-spectrum antimicrobials are urgently required for wound infection management. Robust pre-clinical studies should support the development of such novel antimicrobials.Gap statement. To date, evidence of robust investigation of the cytotoxicity and antimicrobial spectrum of activity of antimicrobial peptides (AMP)s is lacking in published literature. Using a more clinical lens, we address this gap in experimental approach, building on our experience with poly-l-lysine (PLL)-based AMP polymers.Aim. To evaluate the in vitro bactericidal activity and cytotoxicity of a PLL-based 16-armed star AMP polymer, designated 16-PLL10, as a novel candidate antimicrobial.Methods. Antimicrobial susceptibilities of clinical isolates and reference strains of ESKAPE (Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) pathogens, to 16-PLL10 were investigated. Human erythrocyte haemolysis and keratinocyte viability assays were used to assess toxicity. Modifications were made to 16-PLL10 and re-evaluated for improvement.Results. Minimum bactericidal concentration of 16-PLL10 ranged from 1.25 µM to ≥25 µM. At 2.5 µM, 16-PLL10 was broadly bactericidal against ESKAPE strains/wound isolates. Log-reduction in colony forming units (c.f.u.) per millilitre after 1 h, ranged from 0.3 (E. cloacae) to 5.6 (K. pneumoniae). At bactericidal concentrations, 16-PLL10 was toxic to human keratinocyte and erythrocytes. Conjugates of 16-PLL10, Trifluoroacetylated (TFA)-16-PLL10, and Poly-ethylene glycol (PEG)ylated 16-PLL10, synthesised to address toxicity, only moderately reduced cytotoxicity and haemolysis.Conclusions. Due to poor selectivity indices, further development of 16-PLL10 is unlikely warranted. However, considering the unmet need for novel topical antimicrobials, the ease of AMP polymer synthesises/modification is attractive. To support more rational development, prioritising clinically relevant pathogens and human cells, to establish selective toxicity profiles in vitro, is critical. Further characterisation and discovery utilising artificial intelligence and computational screening approaches can accelerate future AMP nanomaterial development.

Current Management of Warm Autoimmune Haemolytic Anaemia and Recent Advancements in Research: Interview with a Key Opinion Leader

Warm autoimmune haemolytic anaemia (wAIHA) is a rare, life-threatening disorder caused by autoantibodies that lead to the premature destruction of erythrocytes (haemolysis). There is currently no licensed targeted therapy for wAIHA. Until recently, there has been little research attention on autoimmune haemolytic anaemias (AIHA), with few developments in the field over the past 20 years. The last 3 years have seen a surge in research interest in wAIHA, with the development of potential new therapies for this rare disorder. For this article, the EMJ conducted an interview in June 2024 with key opinion leader, Bruno Fattizzo, from the University of Milan, and Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy, to raise awareness of wAIHA, and explore recent advancements in research on this disease. Fattizzo, who has a wealth of experience and expertise in the clinical management of wAIHA, provided valuable insights into topics such as the clinical and quality of life (QoL) impact of wAIHA on the patient; current management of wAIHA, including steroids, rituximab, immunosuppressants, splenectomy, anticoagulant prophylaxis, and recombinant erythropoietin; and remaining unmet needs in the disease area. Also discussed were potential future therapies for this autoimmune disorder, including tyrosine kinase inhibitors, neonatal fragment crystallisable receptor (FcRn) inhibitors, and B cell activating factor receptor (BAFF-R) antagonists, and the challenges involved in managing patients with wAIHA. Finally, Fattizzo described the patient experience of living with wAIHA, proposed how best to raise awareness of wAIHA among healthcare professionals, the scientific community, patients, and the public, and outlined what the future of the management of patients with wAIHA might look like.

Construction and characterization of selenium nanoparticles stabilized by cranberry polyphenols with protective effects on erythrocyte hemolysis

Cranberry polyphenols (CPs) were obtained via ultrasound-assisted extraction and purification on a macroporous resin X-5 column. A green synthesis method of selenium nanoparticles (SeNPs) using CPs as reducing agents was then developed. Two types of SeNPs (CP-SeNPs1 and CP-SeNPs2) were successfully constructed and characterized. The spherical particles were well-dispersed on the polyphenol templates and the addition of the polyphenols reduced the aggregation of the nanoparticles. Both CP-SeNPs1 and CP-SeNPs2, with average particle sizes of 85.62 ± 0.11 nm and 107.97 ± 0.12 nm, respectively, demonstrated radical-scavenging activities and protective effects on erythrocyte hemolysis. CP-SeNPs2 possessed more significant antioxidant activity, as evidenced by its higher radical-scavenging rate and greater enhancement of the erythrocyte antioxidant state compared to those of CP-SeNPs1. This study provides a new application of CPs and confirms their great potential in stabilizing SeNPs.

Investigation of Antioxidant Mechanisms of Novel Peptides Derived from Asian Swamp Eel Hydrolysate in Chemical Systems and AAPH-Induced Human Erythrocytes.

Sixteen novel antioxidant peptides from Asian swamp eel (ASE) were identified in previous studies. However, their chemical and cellular antioxidant mechanisms remain unclear. Molecular docking of these peptides with ABTS and DPPH radicals revealed the critical role of hydrogen bonding and Pi-Pi stacking hydrophobic interactions between hydrophobic amino acid residues and free radicals. Residues, such as tryptophan, proline, leucine, and valine, played significant roles in these interactions. All these peptides exhibited notable erythrocyte morphoprotective effects in a model of AAPH-induced oxidative damage of human erythrocytes. Erythrocyte hemolysis was reduced primarily through the modulation of both non-enzymatic (GSH/GSSG) and enzymatic antioxidant systems (SOD, CAT, and GSH-Px) by these peptides. A decrease in levels of MDA, LDH release, and hemoglobin oxidation was observed. Among the peptides, VLYPW demonstrated superior chemical and cellular antioxidant activities, which may be attributed to its higher levels of tyrosine and tryptophan, as well as to its increased hydrophobic amino acid content.

Cu nanowire – impregnated activated carbon fiber for antibacterial applications

Metal nanowires are a 1D structure possessing distinct properties suitable for diverse applications. Copper nanowires (Cu NW) have garnered attention in recent times, particularly as an antibacterial material. Nevertheless, their utilization as a bandage or dressing material necessitates a suitable support, an aspect that remains largely unexplored. In this study, we have developed activated carbon fiber (ACF)-supported Cu NW using the chemical reduction and wet impregnation methods. Tested against Escherichia coli and Staphylococcus aureus, Cu NW/ACF exhibits 83 - 86% reduction in the bacterial growth. The improved antibacterial activity of the material is attributed to the relatively slow release of Cu ions from Cu NW, attributed to the Kelvin effect. Biocompatibility test confirms less than 1.5% hemolysis of erythrocytes, indicating the potential application of the composite as a bandage or dressing material.

Anti-adhesive, anti-biofilm and fungicidal action of newly synthesized gemini quaternary ammonium salts

Newly synthesized gemini quaternary ammonium salts (QAS) with different counterions (bromide, hydrogen chloride, methylcarbonate, acetate, lactate), chain lengths (C12, C14, C16) and methylene linker (3xCH2) were tested. Dihydrochlorides and dibromides with 12 carbon atoms in hydrophobic chains were characterized by the highest biological activity against planktonic forms of yeast and yeast-like fungi. The tested gemini surfactants also inhibited the production of filaments by C. albicans. Moreover, they reduced the adhesion of C. albicans cells to the surfaces of stainless steel, silicone and glass, and slightly to polystyrene. In particular, the gemini compounds with 16-carbon alkyl chains were most effective against biofilms. It was also found that the tested surfactants were not cytotoxic to yeast cells. Moreover, dimethylcarbonate (2xC12MeCO3G3) did not cause hemolysis of sheep erythrocytes. Dihydrochlorides, dilactate and diacetate showed no mutagenic potential.

Purinergic Receptor Antagonists Inhibit Hemolysis Induced by Clostridium perfringens Alpha Toxin.

Clostridium perfringens alpha toxin (CPA), which causes yellow lamb disease in sheep and gas gangrene and food poisoning in humans, is produced by all types of C. perfringens and is the major virulence determinant of C. perfringens type A. CPA induces hemolysis in many species, including humans, murines, sheep and rabbits, through its enzymatic activity, which dissolves the cell membrane. Recent studies have shown that some pore-forming toxins cause hemolysis, which is achieved by the activation of purinergic receptors (P2). However, the relationship between P2 receptors and non-pore-forming toxin hemolysis has not been investigated. In the present study, we examined the function of P2 receptors in CPA toxin hemolysis and found that CPA-induced hemolysis was dependent on P2 receptor activation, and this was also true for Staphylococcus aureus β-Hemolysin, another non-pore-forming toxin. Furthermore, we use selective P2 receptor antagonists to demonstrate that P2X1 and P2X7 play important roles in the hemolysis of human and murine erythrocytes. In addition, we found that redox metabolism was mainly involved in CPA-induced hemolysis using metabolomic analysis. We further demonstrate that CPA activates P2 receptors and then activates NADPH oxidase through the PI3K/Akt and MEK1/ERK1 pathways, followed by the production of active oxygen to induce hemolysis. These findings contribute to our understanding of the pathological effects of CPA, clarify the relationship between P2 activation and non-pore-forming toxin-induced hemolysis, and provide new insights into CPA-induced hemolysis.

Bleomycin‐dependent DNA damage, erythrocyte hemolysis, antitumor MTT assay, and antimicrobial activity studies for Cd (II), Mn (II), Zn (II), Cr (III), and Fe (III) complexes of a multidentate carbohydrazone ligand

Cd (II), Mn (II), Zn (II), Cr (III), and Fe (III) complexes of a carbohydrazone ligand H4EBC, prepared by the reaction of ethyl acetate and carbohydrazide, have been prepared and fully characterized. The ligand H4EBC acted as a bidentate carbohydrazone with Cd (II) and coordinated through the nitrogen of both (‐NH) and (C=N)azomethine groups, while with Mn (II) and Zn (II), it acted as a neutral NN bidentate ligand. In complexes with Fe (III) and Cr (III), H4EBC acts a bi‐negative tetra‐dentate, coordinating through the oxygen atoms of both enolized (C=O)ester groups and the nitrogen of both (C=N)azomethine groups. To illustrate the proposed geometries and chemical reactivity of the compounds, we employed DFT calculations. Additionally, thermal studies provided insights into the stability of the isolated compounds. Interestingly, the prepared complexes demonstrated emission broad bands at various wavelengths, indicating ligand to metal charge transfer. Based on the inhibition of DNA destruction, the complexes can be ranked in the following order: [Cd(H4EBC)Cl2].2H2O] (1) > H4EBC > [Mn(H4EBC)Cl2)].(H2O)2] (2) > [Zn(H4EBC)Cl2].2H2O] (3) > [Fe(H2EBC)Cl(H2O)].H2O (5) > [Cr(H2EBC)Cl (H2O)].H2O (4). Regarding erythrocyte hemolysis, the order of effectiveness can be arranged as follows: (2) > (1) > H4EBC > (5) > (3) > (4). We further investigated the compounds for their potential in antitumor activity against two cell lines and antimicrobial behavior.

INVESTIGATION OF POTENTIAL ACTIVITIES OF PEGANUM HARMALA SEEDS: IN SILICO AND IN VITRO ANALYSES

Seeds of Peganum harmala L. have been traditionally used in Algerian medicine. This study investigates whether the antioxidant, antihemolytic, and anti-inflammatory activities of Peganum harmala extracts (PHE) are attributed to polyphenolic compounds, which are abundant in methanol, chloroform, and ethyl acetate extracts. Extraction and fractionation of polyphenols involved solvents with different polarities, resulting in a crude extract (CrE), a chloroform extract (CHE), and an ethyl acetate extract (EAE). The antioxidant potential of CrE and its fractions was assessed using the ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The inhibition of mice erythrocyte hemolysis was evaluated for methanol, chloroform, and ethyl acetate extracts in the presence of the oxidant (AAPH). PMA-induced mouse ear edema was used as an in vivo model for inflammation. The FTC assay demonstrated the strong antioxidant effect of CrE (87.64 ± 0.003%). EAE showed potent antioxidant activity with low MDA absorption levels. Assessment of antihemolytic effects against AAPH-induced oxidative hemolysis revealed significant protective effects of CrE and EAE, with EAE showing the most pronounced effect. In the in vivo model, CrE (100 mg/kg) exhibited substantial anti-inflammatory activity, inhibiting the ear edema. Computational analyses using molecular docking simulations showed that chlorogenic acid, hesperetin, and rutin have promising potential as inhibitors of COX-2 protein, which is a key component in inflammatory pathways. This study highlights the potent antioxidant properties of P. harmala, particularly in CrE, and its anti-inflammatory effects. The bioactive compounds, such as chlorogenic acid, hesperetin, and rutin, exhibit potential as anti-inflammatory agents. P. harmala could be a valuable natural source for potential medical applications, suggesting the need for further exploration in medical treatments.

Dessecting the toxicological profile of polysorbate 80 (PS80): comparative analysis of constituent variability and biological impact using a zebrafish model

Polysorbate 80, commonly abbreviated as PS80, is a widely used pharmaceutical excipient renowned for its role as a solubilizer and stabilizer in drug formulations. Although PS80 is essential for various pharmaceutical applications, particularly in the formulation of injectable drugs, it has been implicated in a range of adverse reactions. However, due to the complexity of the composition of PS80, the differences in the types and contents of the constituents of PS80 from different manufacturers increase the probability or likelihood of their uneven quality.Addressing the complete spectrum of PS80′s components is challenging; thus, most studies to date have examined PS80 as a singular entity. This approach, however, carries a degree of uncertainty, as it overlooks the unique composition and concentration of components within the PS80 used in experiments, which may not reflect the actual diversity in commercially available PS80 products.Recognizing the critical need to understand how PS80′s composition influences biological effects and toxicity, our study aims to bridge this knowledge gap. By doing so, we can clarify how different PS80 compositions from various manufacturers might affect the quality of pharmaceutical formulations, and also guide excipient manufacturers toward producing higher-quality PS80. Such insights could further facilitate a more targeted application of PS80 in drug development.Building on our previous work, we isolated and prepared two key components of PS80—polyoxyethylene sorbitan monooleate (PSM) and polyoxyethylene isosorbide monooleate (PIM)—and conducted a systematic comparison. We evaluated the acute, hemolytic, and target organ toxicity of two different PS80 samples, as well as PSM and PIM, using a zebrafish model. Our research also delved into the potential mechanisms behind the observed toxicological effects, providing an in-depth understanding of PS80′s impact on biological systems.The results show that PS80, PSM, and PIM resulted in developmental anomalies in larval zebrafish. The primary organs of acute toxicity in zebrafish exposed to PS80 and its typical components PSM and PIM include the cardiovascular system, kidneys, intestines, skin, and liver. Notably, PIM further induced severe pericardial edema and erythrocyte hemolysis, thereby affecting blood flow. The samples also instigated oxidative damage by disrupting the redox equilibrium in the larvae. Compared to PS80, both PSM and PIM induced greater oxidative damage, with PIM notably causing significantly higher lipid oxidation, suggesting that oxidative stress may play a crucial role in polysorbate80-induced toxicity. Furthermore, our study found that PS80 could induce alterations in DNA conformation. The findings underscore the necessity for excipient regulators to establish comprehensive quality standards for Polysorbate 80 (PS80). By implementing such standards, it is possible to minimize the clinical risks associated with the variability in PS80 composition, ensuring safer pharmaceutical products for patients.

Inhibition of LPS-Induced Skin Inflammatory Response and Barrier Damage via MAPK/NF-κB Signaling Pathway by Houttuynia cordata Thunb Fermentation Broth.

Houttuynia cordata Thunb is rich in active substances and has excellent antioxidant and anti-inflammatory activity. Scanning electron microscopy and gel permeation chromatography were used to analyze the molecular characteristics of the fermentation broth of Houttuynia cordata Thunb obtained through fermentation with Clavispora lusitaniae (HCT-f). The molecular weight of HCT-f was 2.64265 × 105 Da, and the polydispersity coefficient was 183.10, which were higher than that of unfermented broth of Houttuynia cordata Thunb (HCT). By investigating the active substance content and in vitro antioxidant activity of HCT-f and HCT, the results indicated that HCT-f had a higher active substance content and exhibited a superior scavenging effect on 2,2-diphenyl-1-picrylhydrazyl radicals and hydroxyl radicals, with IC50 values of 11.85% and 9.01%, respectively. Our results showed that HCT-f could effectively alleviate the increase in the secretion of inflammatory factors and apoptotic factors caused by lipopolysaccharide (LPS) stimulation, and had a certain effect on repairing skin barrier damage. HCT-f could exert an anti-inflammatory effect by down-regulating signaling in the MAPK/NF-κB pathway. The results of erythrocyte hemolysis and chicken embryo experiments showed that HCT-f had a high safety profile. Therefore, this study provides a theoretical basis for the application of HCT-f as an effective ingredient in food and cosmetics.

A xanthan gum and carbomer-codispersed divalent manganese ion-loaded tannic acid nanoparticle adjuvanted inactivated pseudorabies virus vaccine induces balanced humoral and cellular immune responses

Adjuvants including aluminum adjuvant (Alum) and oil-water emulsion have been widely used in inactivated pseudorabies virus (PRV) vaccines to improve their performance, however, they are not sufficient to protect from PRV infection because of the weak immune response and poor Th1-type immune response. Divalent manganese ion (Mn2+) has been reported to increase the cellular immune response significantly. In this work, a xanthan gum and carbomer-dispersed Mn2+-loaded tannic acid-polyethylene glycol (TPMnXC) nanoparticle colloid is developed and used as an adjuvant to improve the performance of the inactivated PRV vaccine. The good in vitro and in vivo biocompatibility of the developed TPMnXC colloid has been confirmed by the cell viability assay, erythrocyte hemolysis, blood routine analysis, and histological analysis of mouse organs and injection site. The TPMnXC-adjuvanted inactivated PRV vaccine (TPMnXC@PRV) significantly promotes higher and more balanced immune responses indicating with an increased specific total IgG antibody and IgG2a/IgG1 ratio, efficient splenocytes proliferation, and elevated Th1- and Th2-type cytokine secretion than those of control groups. Wild PRV challenge experiment is performed using mice as a model animal, achieving a protection rate of up to 86.67 %, which is much higher than those observed from the commercial Alum. This work not only demonstrates the high potentiality of TPMnXC in practical applications but also provides a new way to develop the Mn2+-loaded nanoadjuvant for veterinary vaccines.