Hemolysis Test Research Articles

Tailoring microstructure, mechanical properties, and biocompatibility of Zr alloys via the addition of Ag

This study focuses on the preparation of zirconium (Zr) -silver (Ag) alloys through the alloying of pure Zr with a specific content of Ag using an electric arc furnace. Subsequently, the mechanical and biological properties of these alloys were examined. The analysis conducted, including XRD, OM, SEM, and TEM, confirms the well-established understanding that the Zr–Ag alloy matrix exhibits an α-phase. Furthermore, the addition of Ag elements leads to a gradual refinement of the laths, accompanied by the presence of AgZr2 compounds within the α-Zr matrix. Compression tests demonstrate that the inclusion of Ag components significantly enhances the mechanical properties of the alloys. Notably, the compressive strength of Zr–8Ag reaches an impressive 1932 MPa, while exhibiting an elongation of 22.2 %. Furthermore, analysis conducted using an electrochemical workstation reveals that the self-corrosion potentials of Zr-xAg (where x = 2, 4, 6, 8 at.%) tend to exhibit more positive values compared to pure Zr. Additionally, the corrosion current densities of Zr–6Ag and Zr–8Ag are lower, indicating an improved corrosion resistance. Upon conducting an examination of the antimicrobial test and hemolysis test, it was ascertained that the inclusion of Ag elements significantly augments the antimicrobial characteristics of the alloy. Furthermore, the Zr–Ag alloys exhibit hemolysis rates below 5 %, rendering them appropriate for implementation in biological research endeavors.

Effective photothermal-photodynamic treatment of hepatocellular carcinoma based on polydopamine-coated mesoporous silica nanoparticles

The lack of tumor targeting ability and the ineffectiveness of single treatment modalities are two obstacles in the treatment of hepatocellular carcinoma (HCC). Herein, mesoporous silica nanoparticles (MSNs) with different sizes (MSN100 and MSN50) were synthesized and functionalized with polydopamine (PDA), hypericin B (HB), and arginine-glycine-aspartic acid (RGD) peptides for combined photothermal therapy (PTT) and photodynamic therapy (PDT) of HCC targeting αvβ3 integrin. MSN@PDA-RGD-HB were characterized by various techniques and showed selective drug release, high photothermal conversion efficiency, and reactive oxygen species (ROS) generation under laser irradiation. MSN100@PDA-RGD-HB exhibited higher cellular uptake and HCC toxicity than MSN50@PDA-RGD-HB in vitro, and specifically targeted HCC cells over normal cells. MSN100@PDA-RGD-HB demonstrated high biocompatibility in vivo, as confirmed by the hemolytic test and H&E staining. Importantly, MSN100@PDA-RGD-HB achieved a synergistic effect of PTT and PDT for HCC treatment in mice xenograft tumor model. Therefore, MSN100@PDA-RGD-HB has great potential for tumor therapy in the future.

WITHDRAWN: Preparation, evaluation and application of MRI detectable sunitinib-loaded calcium alginate/poly(acrylic acid) hydrogel microspheres

Transcatheter arterial chemoembolization (TACE) is an effective method for the treatment of unresectable hepatocellular carcinoma. Although many embolic agents have been developed in TACE, there are few ideal embolic agents that combine drug loading, imaging properties and vessel embolization. Here, we developed novel magnetic embolic microspheres that could simultaneously load sunitinib malate (SU), be detected by magnetic resonance imaging (MRI) and block blood vessels. Calcium alginate/poly (acrylic acid) hydrogel microspheres (CA/PAA-MDMs) with superparamagnetic iron oxide nanoparticles (SPIONs) modified by citric acid were prepared by a drip and photopolymerization method. The embolization and imaging properties of CA/PAA-MDMs were evaluated through a series of experiments such as morphology, X-ray diffraction and X-ray photoelectron spectroscopy, magnetic responsiveness analysis, elasticity, cytotoxicity, hemolysis test, in vitro MRI evaluation, rabbit ear embolization and histopathology. In addition, the ability of drug loading and drug release of CA/PAA-MDMs were investigated by using sunitinib (SU) as the model drug. In conclusion, CA/PAA-MDMs showed outstanding drug loading capability, excellent imaging property and embolization effect, which would be expected to be used as a potential biodegradable embolic agent in the clinical interventional therapy.

Development of pH-Sensitive Nanoparticle Incorporated into Dissolving Microarray Patch for Selective Delivery of Methotrexate.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that attacks human joints. Methotrexate (MTX), as one the most effective medications to treat RA, has limitations when administered either orally or by injection. To overcome this limitation, we formulated MTX through a smart nanoparticle (SNP) combined with dissolving microarray patch (DMAP) to achieve selective-targeted delivery of RA. SNP was made using the combination of polyethylene glycol (PEG) and polycaprolactone (PCL) polymers, while DMAP was made using the combination of hyaluronic acid and polyvinylpyrrolidone K-30. SNP-DMAP was then evaluated for its mechanical and chemical characteristics, ex vivo permeation test, in vivo pharmacokinetic study, hemolysis, and hen's egg test-chorioallantoic membrane (HET-CAM) test. The results showed that the characteristics of the SNP-DMAP-MTX formulas meet the requirements for transdermal delivery, with the particle size of 189.09 ±12.30 nm and absorption efficiency of 65.40 ± 5.0%. The hemolysis and HET-CAM testing indicate that this formula was non-toxic and non-irritating. Ex vivo permeation shows a concentration of 51.50 ± 3.20 µg/mL of SNP-DMAP-MTX in PBS pH 5.0. The pharmacokinetic profile of SNP-DMAP-MTX showed selectivity and sustained release compared with oral and DMAP-MTX with values of t1/2 (4.88 ± 0 h), Tmax (8 ± 0 h), Cmax (0.50 ± 0.04 μg/mL), AUC (3.15 ± 0.54 μg/mL.h), and mean residence time (MRT) (9.13 ± 0 h). The developed SNP-DMAP-MTX has been proven to deliver MTX transdermal and selectively at the RA site, potentially avoiding conventional MTX side effects and enhancing the effectiveness of RA therapy.

Abstract 5762: Engineering cell membrane nanoparticlesenhance immunotherapy and radiodynamic therapy for lung adenocarcinoma by promoting cGAS-STING and ferroptosis

Abstract Background: Immunotherapy is a promising approach for lung cancer, but its effectiveness is limited by tumor heterogeneity. Nanoparticle-based targeted drug delivery systems offer advantages such as targeted drug-loading capacity, and hypotoxicity, showing great potential in improving therapeutic efficacy. This study introduces a targeted drug delivery strategy based on immune-activating nanoparticles that achieves precise drugs delivery in tumor areas. Consequently, this approach remodels the microenvironment and induces tumor cytotoxicity. Method: In this study, we first constructed a targeted nanoparticle delivery system FP@CC. The vehicle utilized lung adenocarcinoma cell membranes as the outer shell and encapsulated iron-platinum nanoparticles and STING agonist Vadimezan. Then, TREM2 antibodies were conjugated on the outer layer. The structure of FP@CC-aT2 was characterized by HADDF-STEM and its safety was evaluated by hemolysis test. In vitro, its ability to induce death was verified by AM/PI staining, and to induce ferroptosis was detected by C11 assay. Furthermore, we evaluated its antitumor effects in subcutaneous tumor models. The composition of immune cells within the tumor microenvironment was analyzed using flow cytometry. Results: We initially performed HADDF-STEM analysis on the nanoparticle precursor FP/Vad@CC, revealing its uniform sphericity. For security, no significant hemolysis was observed at different concentrations of FP/Vad@CC. Then, the structures of FP/Vad@CC-aT2 and FP/Vad@CC are relatively stable (Fig 1).The number of dead cells significantly increased after deal with FP@CC, and the effect was significantly enhanced when combined with X-ray. Moreover, C11, utilizing to assess the changes in intracellular reactive oxygen species (ROS), found that FP@CC led to a significant increase in ROS, comparable to induction by H2O2. Both could be effectively prevented by ferroptosis inhibitor(Fig 2).In vivo, FP/Vad@CC-aT2 was applied to a subcutaneous tumor model in C57. It exhibited excellent therapeutic efficacy and biocompatibility. Flow cytometry revealed that FP/Vad@CC-aT2 had the anticipated impact on macrophages within the tumor: a significant reduction in CD11high/CD206high cells and opposite in the ratio of M1/M2-like macrophages (Fig 3). Conclusion: This study developed a targeted drug delivery strategy, FP/Vad@CC-aT2, to improve the tumor microenvironment (TME). We conducted preliminary experiments in vitro and in vivo to validate its physicochemical properties and tumor inhibitory effects, including the induction of ferroptosis. We assessed its safety in mouse and obtained initial evidence of FP/Vad@CC-aT2's ability to remodel the subtypes of macrophage.Moving forward, we will validate its therapeutic and impact on the TME after surgical procedures. Citation Format: Haiyu Zhou, Hongrui Qiu, Shiying Li, Bin Xu, Shengbo Liu, Huili Wang, Hengliang Hou. Engineering cell membrane nanoparticlesenhance immunotherapy and radiodynamic therapy for lung adenocarcinoma by promoting cGAS-STING and ferroptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5762.

Research on strategies for reducing the irritability of potassium cocoyl glycinate in cleansing

BackgroundPotassium cocoyl glycinate, a kind of amino acid surfactant, is widely used as major component for facial cleansing. However, the interaction between surfactants and compounds in the stratum corneum has been suggested as a contributing factor to their potential skin irritancy. ObjectiveThis study aimed to the effects of adding surfactants, polymers, lipids and active substances on diminishing skin irritation caused by potassium cocoyl glycinate. Materials and methodsZein test and Hemolysis test enabled the identification of irritation to proteins and cells in the studied binary systems. The critical micelle concentration (CMC) was evaluated by the surface tension method. The power of the analyzed polymers to diminish irritation of potassium cocoyl glycinate was represented by the reduction rate. Results(1) Amphoteric surfactants and nonionic surfactants are effective at reducing the irritation of potassium cocoyl glycinate in some cases by reducing CMC; (2) Addition of polymers to the potassium cocoyl glycinate solutions improves its safety of use. Hydroxypropyl methylcellulose could reduce the irritation by 88.63%; (3) Combining most lipids can assist in declining the irritation, Lppg-28-Buteth-35 could inhibit the irritation of red blood cells by 16.52%; (4) The effects of active substances varied, hydrolyzed sodium hyaluronate results in a significant reduction in irritation to cells. ConclusionIn facial cleansers based on potassium cocoyl glycinate, supplying co-surfactant, polymers, lipids and active substances can reduce the irritation to some extent, however, the effects are various.

Challenging the dogma: Red blood cell-directed autoimmunity as risk factor for red blood cell alloimmunisation after blood transfusion.

Red blood cell autoimmunity and alloimmunity are potentially linked. Quantification of this association can tailor extensively matched red blood cell transfusions in patients with autoimmunity. Using an incident new-user cohort comprising 47 285 previously non-transfused, non-alloimmunised patients, we compared transfusion-induced red blood cell alloimmunisation incidences in direct antiglobulin test (DAT)-positive and control patients. Additionally, we performed case-control analyses to handle potential confounding by clinical immunomodulators. Among (IgG and/or C3d) DAT-positive patients (N = 380), cumulative red blood cell alloimmunisation incidences after 10 units transfused reached 4.5% (95% confidence interval [CI] 2.5-8.2) versus 4.2% (CI 3.9-4.5, p = 0.88) in controls. In case-control analyses, alloimmunisation relative risks among DAT-positive patients increased to 1.7 (CI 1.1-2.8). Additional adjustments for pre-DAT transfusion exposure or the extent of Rh/K mismatching did not impact results. In conclusion, while patients with DAT positivity show an intrinsically increased alloimmune red blood cell response, their absolute risk is comparable to control patients due to counteracting co-existing immunosuppressive conditions. Consequently, isolated DAT positivity in patients lacking overt haemolysis or complicated alloantibody testing does not seem to warrant extended matching strategies.

Carrageenan-xanthan nanocomposite film with improved bioadhesion and permeation profile in human skin: A cutaneous-friendly platform for ketoprofen local delivery

Ketoprofen (KET), commonly used for inflammation in clinical settings, leads to systemic adverse effects with prolonged use, mitigated by topical administration. Nanotechnology-based cutaneous forms, like films, may enhance KET efficacy. Therefore, this study aimed to prepare and characterize films containing KET nanoemulsions (F-NK) regarding mechanical properties, chemical composition and interactions, occlusive potential, bioadhesion, drug permeation in human skin, and safety. The films were prepared using a κ-carrageenan and xanthan gum blend (2 % w/w, ratio 3: 1) plasticized with glycerol through the solvent casting method. Non-nanoemulsioned KET films (F-K) were prepared for comparative purposes. F-NK was flexible and hydrophilic, exhibited higher drug content and better uniformity (94.40 ± 3.61 %), maintained the NK droplet size (157 ± 12 nm), and was thinner and lighter than the F-K. This film also showed increased tensile strength and Young's modulus values, enhanced bioadhesion and occlusive potential, and resulted in more of the drug in the human skin layers. Data also suggested that nano-based formulations are homogeneous and more stable than F-KET. Hemolysis and chorioallantoic membrane tests suggested the formulations' safety. Thus, the nano-based film is suitable for cutaneous KET delivery, which may improve the drug's efficacy in managing inflammatory conditions.

A novel approach to wound healing: Green synthetic nano-zinc oxide embedded with sodium alginate and polyvinyl alcohol hydrogels for dressings

Wound healing constitutes a formidable challenge within the healthcare system, attributable to infection risks and protracted recovery periods. The pressing need for innovative wound healing methods has spurred the urgency to develop novel approaches. This study sought to advance wound healing by introducing a novel approach employing a composite sponge dressing. The composite sponge dressing, derived from LFL-ZnO (synthesized through the green methodology utilizing Lactobacillus plantarum ZDY2013 fermentation liquid), polyvinyl alcohol (PVA), and sodium alginate (SA) via a freeze–thaw cycle and freeze-drying molding process, demonstrated notable properties. The findings elucidate the commendable swelling, moisturizing, and mechanical attributes of the SA/LFL-ZnO/PVA composite sponge dressing, characterized by a porous structure. Remarkably, the dressing incorporating LFL-ZnO exhibited substantial inhibition against both methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus (S. aureus). Hemolysis and cytotoxicity tests corroborated the excellent biocompatibility of the sponge dressing. In vivo evaluation of the therapeutic efficacy of the 1 mg/mL LFL-ZnO composite dressing on scald wounds and S. aureus-infected wounds revealed its capacity to accelerate wound healing and exert pronounced antibacterial effects. Consequently, the composite sponge dressings synthesized in this study hold significant potential for application in wound treatment.

Quinalizarin as a potential antifungal drug for the treatment of Candida albicans fungal infection in cancer patients.

This study aims to analyze the antifungal properties of quinalizarin, a plant-derived compound with proven anticancer effects. Quinalizarin exhibited antifungal activity against opportunistic pathogenic Candida species and Geotrichum capitatum. The treatment with this anthraquinone reduced hyphal growth, inhibited biofilm formation, and damaged mature Candida albicans biofilms. Real-time RT-PCR revealed that quinalizarin downregulated the expression of hyphae-related and biofilm-specific genes. The flow cytometry method used in the study showed that both apoptosis and necrosis were the physiological mechanisms of quinalizarin-induced C. albicans cell death, depending on the dose of the antifungal agent. A further study revealed an increase in the levels of intracellular reactive oxygen species and alterations in mitochondrial membrane potential after treatment with quinalizarin. Finally, quinalizarin was found to have low toxicity in a hemolytic test using human erythrocytes. In conclusion, we have identified quinalizarin as a potential antifungal compound.IMPORTANCEThis article is a study to determine the antifungal activity of quinalizarin (1,2,5,8-tetrahydroxyanthraquinone). Quinalizarin has potential antitumor properties and is effective in different types of tumor cells. The aim of the present study was to prove that quinalizarin can be used simultaneously in the treatment of cancer and in the treatment of intercurrent fungal infections. Quinalizarin was identified as a novel antifungal compound with low toxicity. These results may contribute to the development of a new drug with dual activity in the treatment of cancer-associated candidiasis.

The time course of complement blockage by ravulizumab in neuromyelitis optica spectrum disorder

AbstractThe classical 50% hemolytic complement test is commonly used to monitor complement function. However, its use in monitoring the effect of ravulizumab on complement assays is unknown. A 47‐year‐old man with anti‐aquaporin 4 antibody‐positive neuromyelitis optica spectrum disorder who was treated with oral prednisolone and azathioprine experienced several episodes of optic neuritis or myelitis. After his latest relapse, ravulizumab was administered to prevent relapse. Serum classical 50% hemolytic complement activity decreased below detectable levels (<10 U/mL) within 1 h of ravulizumab injection. Eighteen weeks later, the patient showed no adverse events or relapse, suggesting that serum classical 50% hemolytic complement activity can monitor ravulizumab effect. In Japan, where 3.5% of the population shows a reduced response to anti‐C5 therapy, serum classical 50% hemolytic complement activity proves useful in confirming ravulizumab efficacy.

Characterization of endophytic bacteria of Mindi (Melia azedarach) and Balsa (Ochroma pyramidale) plants and potential inhibition in plant diseases

Plant diseases attack all parts of the plant, such as leaves, stems, and roots. The cause of pest attacks is due to several factors, namely environmental conditions that support pathogens, selected plant conditions that are susceptible to disease, and poor maintenance. Disease attacks on the roots include attacks that are harmful to plants, so it is necessary to have effective control carried out by considering the ecology and economy. One of the techniques for controlling plant-disturbing organisms (OPT) can be carried out by utilizing endophytic microbes. This study aimed to explore the types of endophytic bacteria found in balsa and mindi, which have the potential as biological control agents, as well as the effect of using endophytic bacteria in suppressing the growth of root diseases in vitro. The study found that as many as 20 endophytic bacteria were purified from balsa, and 25 individuals were purified from mindi. The hemolysis test and plant hypersensitivity test results showed seven pure isolates with endophytic potential, namely three isolates from balsa and four from mindi.

Formulation and evaluation of dissolving microneedle for transdermal delivery of piperine: the effect of polymers concentration

This research aims to develop the formulation of Dissolving Microneedle Piperine (DMNs PIP) and evaluate the effect of polymer concentration on characterisation and permeation testing results in ex vivo. DMNs PIP were prepared from varying concentrations of piperine (PIP) (10, 15, and 20% w/w) and polymers of polyvinyl alcohol (PVA): Polyvinyl pyrrolidone (30:60 and 60:25), respectively. Then the morphological evaluation of the formula was carried out, followed by mechanical strength testing. Furthermore, the density, LOD, and weight percentage of piperine in the dried microneedle were calculated and the determination of volume, needle weight and piperine weight and analysed. Ex vivo testing, X-Ray Diffraction, FTIR and hemolysis tests were carried out. PIP with PVA and PVP (F1) polymers produced DMN with mechanical strength (8.35 ± 0.11%) and good penetration ability. In vitro tests showed that the F1 polymer mixture gave good penetration (95.02 ± 1.42 μg/cm2), significantly higher than the F2, F3, F4, and F5 polymer mixtures. The DMNs PIP characterisation results through XRD analysis showed a distinctive peak in the 20–30 region, indicating the presence of crystals. The FTIR study showed that the characteristics of piperine found in DMNs PIP indicated that piperine did not undergo interactions with polymers. The results of the ex vivo study through DMNs PIP hemolytic testing showed no hemolysis occurred, with the hemolysis index below the 5% threshold reported in the literature. These findings indicate that DMNs PIP is non-toxic and safe to use as alternative for treating inflammation

Targeted Screening of Fiber Degrading Bacteria with Probiotic Function in Herbivore Feces.

Cellulolytic bacteria with probiotic functions play a crucial role in promoting the intestinal health in herbivores. In this study, we aimed to correlate the 16S rRNA gene amplicon sequencing and fiber-degrading enzyme activity data from six different herbivore feces samples. By utilizing the separation and screening steps of probiotics, we targeted and screened high-efficiency fiber-degrading bacteria with probiotic functions. The animals included Maiwa Yak (MY), Holstein cow (CC), Tibetan sheep (TS), Southern Sichuan black goat (SG), Sichuan white rex rabbit (CR), and New Zealand white rabbit (ZR). The results showed that the enzymes associated with fiber degradation were higher in goat and sheep feces compared to cattle and rabbit's feces. Correlation analysis revealed that Bacillus and Fibrobacter were positively correlated with five types of fiber-degrading related enzymes. Notably, the relative abundance of Bacillus in the feces of Tibetan sheep was significantly higher than that of other five herbivores. A strain TS5 with good cellulose decomposition ability from the feces of Tibetan sheep by Congored staining, filter paper decomposition test, and enzyme activity determination was isolated. The strain was identified as Bacillus velezensis by biological characteristics, biochemical analysis, and 16S rRNA gene sequencing. To test the probiotic properties of Bacillus velezensis TS5, we evaluated its tolerance to acid and bile salt, production of digestive enzymes, antioxidants, antibacterial activity, and adhesion ability. The results showed that the strain had good tolerance to pH 2.0 and 0.3% bile salts, as well as good potential to produce cellulase, protease, amylase, and lipase. This strain also had good antioxidant capacity and the ability to antagonistic Staphylococcus aureus BJ216, Salmonella SC06, Enterotoxigenic Escherichia coli CVCC196, and Escherichia coli ATCC25922. More importantly, the strain had good self-aggregation and Caco-2 cell adhesion rate. In addition, we tested the safety of Bacillus velezensis TS5 by hemolysis test, antimicrobial susceptibility test, and acute toxicity test in mice. The results showed that the strain had no hemolytic phenotype, did not develop resistance to 19 commonly used antibiotics, had no cytotoxicity to Caco-2, and did not have acute toxic harm to mice. In summary, this study targeted isolated and screened a strain of Bacillus velezensis TS5 with high fiber-degrading ability and probiotic potency. This strain can be used as a potential probiotic for feeding microbial preparations for ruminants.

Preparation of Folic acid@Arsenic nanoparticles and evaluation of their antioxidant properties and cytotoxic effects.

In this study, arsenic nanoparticles containing folic acid (FA@As NPs) were synthesized by microwave irradiating a mixture of As2O3 and sodium borohydride solution in the presence of folic acid. The physicochemical characteristics of the prepared NPs were studied by UV-visible spectroscopy, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) analyses. Antioxidant activities, hemocompatibility, and cytotoxic effects of the prepared NPs were then evaluated. The attained results showed that the hexagonal FA@As NPs have a size range between 12.8 nm and 19.5 nm. The DPPH scavenging activity of FA@As NPs was found to be significantly greater than that of As NPs at concentrations ranging from 40 µg/mL to 2560 µg/mL (p<0.05). The hemolytic test confirmed that the measured hemolysis percentage (HP) for FA@As NPs and As NPs was 0% at concentrations between 20 to160 µg/mL, and for FA@As NPs, the measured HP was not significantly higher than As NPs at concentrations higher than 320 µg/mL (p>0.05). The necessary concentration for the death of half of the cells (IC50) for MDA-MB-231, MCF-7, and HUVEC cells treated (24 h) with FA@As NPs was measured to be 19.1±1.3 µg/mL, 15.4±1.1 µg/mL, and 16.8±1.2 µg/mL, respectively. However, further investigations are necessary to clarify the mechanisms behind the biological activities of FA@As NPs.

Rational Design of Antifungal Peptides Based on the γ-Core Motif of a Neosartorya (Aspergillus) fischeri Antifungal Protein to Improve Structural Integrity, Efficacy, and Spectrum.

Antifungal peptides offer promising alternative compounds for the treatment of fungal infections, for which new antifungal compounds are urgently needed. Constant and broad antifungal spectra of these peptides play essential roles in their reliable therapeutic application. It has been observed that rationally designed peptides using the evolutionarily conserved γ-core region (GXC-X3-9-C) of an antifungal protein from Neosartorya (Aspergillus) fischeri highly inhibit the growth of fungi. The cysteines in these peptides have free sulfhydryl groups, which allow cyclization and dimerization under oxidative conditions, thereby impairing antifungal efficacy. To overcome this problem, one or two cysteine residues were substituted by serines or S-tert-butyl was applied as a cysteine-protecting group. Furthermore, structural integrity and antifungal efficacy investigations before and after oxidative exposure revealed that substituting both cysteines with serines and S-tert-butylation helped maintain the structural integrity. However, it slightly decreased the antifungal efficacy against a yeast, Candida albicans. Interestingly, S-tert-butylation maintained the efficacy and could extend the antifungal activity to a mold, Aspergillus fumigatus. Usually, cyclization and dimerization did not influence the antifungal efficacy of most peptides. Additionally, hemolysis tests and Galleria mellonella toxicity model experiments indicated that none of the applied modifications made the peptides harmful to animals.

Distearoylphosphatidylglyceroland dioleoyl phosphatidylglycerol increase the retention and reduce the toxicity of amphotericin B-loaded in nanoemulsions.

Aim: To develop nanoemulsions (NEs) loading amphotericin B (AmB) and to evaluate the influence of different excipients on the stability and the supramolecular organization, retentionand toxicity of AmB. Materials & methods: The NEs were developed from different oils, surfactants, external mediaand anionic lipids (disteaoryl phosphatidylglycerol [DSPG]and dioleoyl phosphatidylglycerol [DOPG]). Their impact on the size, pH, zeta potential, AmB encapsulation efficiency, AmB retentionand hemolytic potential of the NEs was evaluated. Results& conclusion: The use of soybean oil (lipid matrix), Span 80 (surfactant), phosphate buffer (external phase) and DSPG or DOPG (hydrophobic ion pair) provided better NE stability, higher AmB retention within the NEs and a safer formulationprofile in hemolysis tests.

Azomycin Orchestrate Colistin-Resistant Enterobacter cloacae Complex's Colistin Resistance Reversal In Vitro and In Vivo.

The Enterobacter cloacae complex (ECC) is a group of nosocomial pathogens that pose a challenge in clinical treatment due to its intrinsic resistance and the ability to rapidly acquire resistance. Colistin was reconsidered as a last-resort antibiotic for combating multidrug-resistant ECC. However, the persistent emergence of colistin-resistant (COL-R) pathogens impedes its clinical efficacy, and novel treatment options are urgently needed. We propose that azomycin, in combination with colistin, restores the susceptibility of COL-R ECC to colistin in vivo and in vitro. Results from the checkerboard susceptibility, time-killing, and live/dead bacterial cell viability tests showed strong synergistic antibacterial activity in vitro. Animal infection models suggested that azomycin-colistin enhanced the survival rate of infected Galleria mellonella and reduced the bacterial load in the thighs of infected mice, highlighting its superior in vivo synergistic antibacterial activity. Crystal violet staining and scanning electron microscopy unveiled the in vitro synergistic antibiofilm effects of azomycin-colistin. The safety of azomycin and azomycin-colistin at experimental concentrations was confirmed through cytotoxicity tests and an erythrocyte hemolysis test. Azomycin-colistin stimulated the production of reactive oxygen species in COL-R ECC and inhibited the PhoPQ two-component system to combat bacterial growth. Thus, azomycin is feasible as a colistin adjuvant against COL-R ECC infection.

Biomimetic mineralization of platelet lysate/oxidized dextran cryogel as a macroporous 3D composite scaffold for bone repair

Scaffold development approaches using autologous sources for tissue repair are of great importance in obtaining bio-active/-compatible constructs. Platelet-rich plasma (PRP) containing various growth factors and platelet lysate (PL) derived from PRP are autologous products that have the potential to accelerate the tissue repair response by inducing a transient inflammatory event. Considering the regenerative capacity of PRP and PL, PRP/PL-based scaffolds are thought to hold great promise for tissue engineering as a natural source of autologous growth factors and a provider of mechanical support for cells. Here, a bio-mineralized PRP-based scaffold was developed using oxidized dextran (OD) and evaluated for future application in bone tissue engineering. Prepared PL/OD scaffolds were incubated in simulated body fluid (SBF) for 7, 14 and 21 d periods. Mineralized PL/OD scaffolds were characterized using Fourier transform infrared spectroscopy, x-ray diffraction spectroscopy, scanning electron microscopy (SEM), thermogravimetric analysis, porosity and compression tests. SEM and energy-dispersive x-ray spectroscopy analyses revealed mineral accumulation on the PL/OD scaffold as a result of SBF incubation. In vitro cytotoxicity and in vitro hemolysis tests revealed that the scaffolds were non-toxic and hemocompatible. Additionally, human osteoblasts (hOBs) exhibited good attachment and spreading behavior on the scaffolds and maintained their viability throughout the culture period. The alkaline phosphatase activity assay and calcium release results revealed that PL/OD scaffolds preserved the osteogenic properties of hOBs. Overall, findings suggest that mineralized PL/OD scaffold may be a promising scaffold for bone tissue engineering.

A novel flavonol-polysaccharide from Tamarix chinensis alleviates influenza A virus-induced acute lung injury. Evidences for its mechanism of action

BackgroundTamarix chinensis Lour. is a Chinese medicine used for treating inflammation-related diseases and its crude polysaccharides (MBAP90) exhibited significant anticomplement activities in vitro. PurposeTo obtain anticomplement homogenous polysaccharides from MBAP90 and explore its therapeutic effects and potential mechanism on influenza A virus (IAV)-induced acute lung injury (ALI). MethodsAnticomplement activity-guided fractionation of the water-soluble crude polysaccharides from the leaves and twigs of T. chinensis were performed by diethylaminoethyl-52 (DEAE-52) cellulose and gel permeation columns to yield a homogeneous polysaccharide MBAP-5, which was further characterized using ultra-high-performance liquid chromatography-ion trap tandem mass spectrometry (UPLC-IT-MS) and nuclear magnetic resonance (NMR) analysis. In vitro, the anticomplement activity of MBAP-5 through classical pathway was measured using a hemolytic test. The therapeutic effects of MBAP-5 on ALI were evaluated in H1N1-infected mice. H&E staining, enzyme linked immunosorbent assay (ELISA), immunohistochemistry, and western blot were used to systematically access lung histomorphology, inflammatory cytokines, degree of complement component 3c, 5aR, and 5b-9 (C3c, C5aR, and C5b-9) deposition, and inflammasome signaling pathway protein expressions in lung tissues. ResultsMBAP-5 was a novel flavonol-polysaccharide with the molecular weight (Mw) of 153.6 kDa. Its structure was characterized to process a backbone of →4)-α-D-GlcpA-(1→, →6)-α-D-Glcp-(1→, →3,4)-α-D-Glcp-(1→, →3,4,6)-α-D-Glcp-(1→, and →4,6)-β-D-Glcp-(1→, as well as branches of α-L-Araf-(1→ and β-D-Galp-(1→. Particularly, O-3 of →3,4,6)-α-D-Glcp-(1→ was substituted by quercetin. In vitro assay showed that MBAP-5 had a potent anticomplement activity with a CH50 value of 102 ± 4 µg/ml. Oral administration of MBAP-5 (50 and 100 mg/kg) effectively attenuated the H1N1-induced pulmonary injury in vivo by reducing pulmonary edema, virus replication, and inflammatory responses. Mechanistically, MBAP-5 inhibited the striking deposition and contents of complement activation products (C3c, C5aR, and C5b-9) in the lung. Toll-like receptor 4 (TLR4) /transcription factor nuclear factor κB (NF-κB) signaling pathway was constrained by MBAP-5 treatment. In addition, MBAP-5 could suppress activation of the inflammasome pathways, including Nod‐like receptor pyrin domain 3 (NLRP3), cysteinyl aspartate specific proteinase-1/12 (caspase-1/12), apoptosis‑associated speck‑like protein (ASC), gasdermin D (GSDMD), interleukin (IL)-1β, and IL-18 expressions. ConclusionsA novel flavonol-polysaccharide MBAP-5 isolated from T. chinensis demonstrated a therapeutic effect against ALI induced by IAV attack. The mechanism might be associated with inhibition of complement system and inflammasome pathways activation.