Hemolysis Of Red Blood Cells Research Articles

Investigation of chemical constituents, antioxidant, anti-inflammatory and nutritional properties of oil of Persea americana (Avocado) seeds

This study investigated the phytochemical composition, in vitro antioxidant, and anti-inflammatory properties of the oil of Persea americana seeds (OPAS). The flavonoid and total phenol compositions were quantified using standard colorimetric methods; the antioxidant potentials of the seed were determined using 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, while the anti-inflammatory potentials were investigated using membrane stabilization (hypotonicity) and heat-induced hemolysis of human red blood cells (HRBC) assays. In addition, gas chromatography-mass spectrometry (GC–MS) analysis was employed to characterize the phytochemicals in the oil. The nutritional indices (atherogenic index, thrombogenic index, and polyene index) were also calculated. Results indicated high total phenolic and flavonoid contents (25.78 ± 0.68 mg gallic acid equivalent per gram (GAE/g) and 72.69 ± 1.11 mg quercetin equivalent per 100 mg (QE/100 mg), respectively). The oil of P. americana seeds showed dose-dependent (25–400 μg/mL) antioxidant and anti-inflammatory activities compared to the standard agents used (olive oil and ascorbic acid). The GC–MS analysis showed the presence of fatty acids including tetradecanoic acid (33.33 %), 9, 12-octadecadienoic acid, methyl ester (20.12 %), n-hexadecenoic acid (7.07 %), and oleic acid (11.60 %). Nutritional indices showed that OPAS had a very low polyunsaturated/saturated (PUFA/SFA) ratio (0.45), hypocholesterolemic/hypercholesterolemic (0.26), but high atherogenicity index (IA) (4.13) and thrombogenicity index IT (9.47) values. The beneficial potentials of OPAS revealed in this study suggest that it can be harnessed as a potent antioxidant and anti-inflammatory agent in phytomedicine. However, the nutritional quality and culinary suitability of OPAS with different oils require further study.

Chemical profiling and bioactivity studies on aerial parts Ammoides atlantica (Coss. et Durieu) H. Wolff

The Algerian endemic plant Ammoides atlantica (Coss. et Durieu) H. Wolff was studied for the chemical profiling and biological activities of its essential oil (EO) and ethanolic extract (EE). The chemical analysis by GC/MS and HPLC/DAD/UV revealed, respectively, the major compounds thymol (39.46%), γ-terpinene (31.74%), and p-cymene (19.01%) in the EO, and apigenin (33.58%), luteolin 7-O-glucoside (20.09%), and luteolin (14.39%) in the EE. The EO exhibited strong antioxidant activity, with a significant ABTS•+ scavenging capacity (IC50 = 2.79 µg/mL) compared to EE, Trolox, and BHT. The EE showed comparable effects to BHT in DPPH scavenging and reducing power tests. Moreover, the EO demonstrated noteworthy antibacterial activity against S. aureus, E. coli, and P. aeruginosa, with inhibition zone diameters ranging from 32.1 to 70 mm and MICs below 0.3 to 5 mg/mL. Furthermore, the EE exhibited strong anti-inflammatory activity by inhibiting hemolysis of red blood cells >70% at a concentration of 20 µg/mL.

Ginsenoside Rh2 Regulates the Calcium/ROS/CK1α/MLKL Pathway to Promote Premature Eryptosis and Hemolysis in Red Blood Cells.

Ginsenoside Rh2 (GRh2) exhibits significant potential as an anticancer agent; however, progress in developing chemotherapeutic drugs is impeded by their toxicity toward off-target tissues. Specifically, anemia caused by chemotherapy is a debilitating side effect and can be caused by red blood cell (RBC) hemolysis and eryptosis. Cells were exposed to GRh2 in the antitumor range and hemolytic and eryptotic markers were examined under different experimental conditions using photometric and cytofluorimetric methods. GRh2 caused Ca2+-independent, concentration-responsive hemolysis in addition to disrupted ion trafficking with K+ and Cl- leakage. Significant increases in cells positive for annexin-V-fluorescein isothiocyanate, Fluo4, and 2,7-dichlorofluorescein were noted upon GRh2 treatment coupled with a decrease in forward scatter and acetylcholinesterase activity. Importantly, the cytotoxic effects of GRh2 were mitigated by ascorbic acid and by blocking casein kinase 1α (CK1α) and mixed lineage kinase domain-like (MLKL) signaling. In contrast, Ca2+ omission, inhibition of KCl efflux, and isosmotic sucrose aggravated GRh2-induced RBC death. In whole blood, GRh2 selectively targeted reticulocytes and lymphocytes. Altogether, this study identified novel mechanisms underlying GRh2-induced RBC death involving Ca2+ buildup, loss of membrane phospholipid asymmetry and cellular volume, anticholinesterase activity, and oxidative stress. These findings shed light on the hematologic toxicity of GRh2 which is crucial for optimizing its utilization in cancer treatment.

Rejuvenation of Meropenem by Conjugation with Tilapia Piscidin-4 Peptide Targeting NDM-1 Escherichia coli.

Gram-negative pathogens that produce β-lactamases pose a serious public health threat as they can render β-lactam antibiotics inactive via hydrolysis. This action contributes to the waning effectiveness of clinical antibiotics and creates an urgent need for new antimicrobials. Antimicrobial peptides (AMPs) exhibiting multimodal functions serve as a potential source in spite of a few limitations. Thus, the conjugation of conventional antibiotics with AMPs may be an effective strategy to leverage the advantages of each component. In this study, we conjugated meropenem to the AMP Tilapia piscidin 4 (TP4) using a typical coupling reaction. The conjugate was characterized by using HPLC-MS, HR-MS, and MS-MS fragmentation analysis. It was then evaluated in terms of antibacterial potency, hemolysis, and cytotoxicity toward RAW264.7 and CCD-966SK cell lines. The conjugation of meropenem with TP4 significantly reduced the cytotoxicity compared to TP4. Conjugation of unprotected TP4 with meropenem resulted in cross-linking at the N-terminal and lysine sites. The structural activity relationship of the two isomers of the TP4-meropenem conjugate was investigated. Both the isomers showed notable antibacterial activities against NDM-1 Escherichia coli and reduced red blood cell hemolysis as compared to TP4. Lysine conjugate (TP4-K-Mero) showed lesser hemolysis than the N-terminal conjugate (TP4-N-Mero). Molecular modeling further revealed that the conjugates can bind to lipopolysaccharides and inhibit NDM-1 β-lactamase. Together, these data show that conjugation of antibiotics with AMP can be a feasible approach to increase the therapeutic profile and effectively target multidrug-resistant pathogens. Furthermore, antibiotic conjugation at different AMP sites tends to show unique biological properties.

Biological and Genetic Determinants of Glycolysis: Phosphofructokinase Isoforms Boost Energy Status of Stored Red Blood Cells and Transfusion Outcomes.

Blood donor age and sex affect glycolysis in stored RBCs from 13,029 volunteers;Ancestry, genetic polymorphisms in PFKP, HK1, CD38/BST1 influence RBC glycolysis;Modeled PFKP effects relate to preventing loss of the total AXP pool in stored RBCs;ATP and hypoxanthine are biomarkers of hemolysis in vitro and in vivo.

Ferroptosis regulates hemolysis in stored murine and human red blood cells.

Steap3 regulates lipid peroxidation and extravascular hemolysis in 350 diversity outbred miceSteap3 SNPs are linked to RBC iron, hemolysis, vesiculation in 13,091 blood donorsmQTL analyses of oxylipins identified ferroptosis-related gene products FADS1/2, EPHX2, LPCAT3Ferroptosis markers are linked to hemoglobin increments in transfusion recipients.

Antimicrobial and Membrane Stabilization Activity of Plant Extracts Against Pathogenic Bacteria and Fungi

The use and search for antimicrobial drugs derived from plants have accelerated in recent years. The present study was aimed to determine the antimicrobial, antibiofilm and membrane stabilization activities of plant extracts. Crude extracts of four plants namely, Mikania scandens (L.), Mimosa pudica (L.), Murraya paniculata (L.), and Syzygium aromaticum (L.) were tested against eleven human pathogens including four biofilm producing bacterial strains Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus, and one fungal strain Candida albicans. The antimicrobial activities as well as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extracts were evaluated using disc diffusion and macro-broth dilution methods respectively. Among the plant extracts, ethanol extract of S. aromaticum exhibited the largest zone of inhibition 35 mm in diameter against Bacillus subtilis at 500 µg/disc concentrations. The lowest MIC (1000 µg/ml) and MBC (2500 µg/ml) were determined against P. aeruginosa with the same extract of S. aromaticum. In case of time kill assay, S. aromaticum extract showed the lowest optical density (OD600) 0.2 against E coli in 3h at MBC concentration. Moreover, the same extract of S. aromaticum displayed the strong antibiofilm activity, inhibiting 100% biofilm formation of E. coli at 2×MIC concentration. Furthermore, in vitro experiment was performed to evaluate membrane stabilization activity of plant extracts and S. aromaticum showed 100% activity in stabilizing cell membranes, preventing hemolysis of red blood cells. Therefore, this study provides valuable insight for designing antimicrobial products to efficiently eliminating human infections, and the plant extracts could be the potential antimicrobial agent. Bangladesh J Microbiol, Volume 40, Number 2, December 2023, pp 76-85

“Dark and bright sides” of Gilbert’s syndrome

Gilbert’s syndrome, also known as benign hyperbilirubinemia, was described more than 100 years ago. It has usually been considered a physiological abnormality characterized by a mild elevation of the systemic level of unconjugated bilirubin, in the absence of any underlying liver or overt RBC hemolysis. The molecular basis of Gilbert’s syndrome lies in the impairment of the conjugation of bilirubin with glucuronic acid in the hepatocytes, which is mediated by a specific hepatic enzyme named bilirubin-UDP-glucuronosyl transferase 1A1 that forms bilirubin diglucuronoside. Clearance of various xenobiotics, which are not substrates for glucuronosylation, is impaired in patients with Gilbert’s syndrome; their detailed list is provided in the article. Fatigue, asthenia, and various vaguely defined dyspeptic complaints attributed to Gilbert’s syndrome in the past are no longer considered a part of this condition, and proper evaluation of possible causes is required in these cases. Since the re-discovery of the potent antioxidant effects of bilirubin in the late 1980s, as well as the multiple intracellular signalling pathways affected by bilirubin, an ever-increasing body of evidence suggests that individuals with Gilbert’s syndrome may benefit from the mild hyperbilirubinemia and are actually protected from the development of a wide range of “diseases of civilization”, such as cardiovascular diseases, certain cancers, and autoimmune or neurodegenerative diseases. Gilbert’s syndrome is defined phenotypically, and therefore not according to predisposing genetic markers, as the elevation of serum unconjugated bilirubin concentration above the upper limit of normal, with no laboratory signs of hemolysis or liver damage. This review analyses the current state of medical knowledge given recent discoveries in this rapidly developing field, as well as their possible clinical significance, and provides a new perspective on this condition.

Exploring the Cryopreservation Mechanism and Direct Removal Strategy of TAPS in Red Blood Cell Cryopreservation.

Cryopreservation of red blood cells (RBCs) plays an indispensable role in modern clinical transfusion therapy. Researchers are dedicated to finding cryoprotectants (CPAs) with high efficiency and low toxicity to prevent RBCs from cryopreservation injury. This study presents, for the first time, the feasibility and underlying mechanisms of a novel CPA called tris(hydroxymethyl)aminomethane-3-propanesulfonic acid (TAPS) in RBCs cryopreservation. The results demonstrated that the addition of TAPS achieved a post-thaw recovery of RBCs at 79.12 ± 0.67%, accompanied by excellent biocompatibility (above 97%). Subsequently, the mechanism for preventing RBCs from cryopreservation injury was elucidated. On one hand, TAPS exhibits a significant amount of bound water and effectively inhibits ice recrystallization, thereby reducing mechanical damage. On the other hand, TAPS demonstrates high capacity to scavenge reactive oxygen species and strong endogenous antioxidant enzyme activity, providing effective protection against oxidative damage. Above all, TAPS can be readily removed through direct washing, and the RBCs after washing showed no significant differences in various physiological parameters (SEM, RBC hemolysis, ESR, ATPase activity, and Hb content) compared to fresh RBCs. Finally, the presented mathematical modeling analysis indicates the good benefits of TAPS. In summary, TAPS holds potential for both research and practical in the field of cryobiology, offering innovative insights for the improvement of RBCs cryopreservation in transfusion medicine.

The search for pyruvate kinase-R activators; from a HTS screening hit via an impurity to the discovery of a lead series

Pyruvate kinase (PK) is an essential component of cellular metabolism, converting ADP and phosphoenolpyruvate (PEP) to pyruvate in the final step of glycolysis. Of the four unique isoforms of pyruvate kinase, R (PKR) is expressed exclusively in red blood cells and is a tetrameric enzyme that depends on fructose-1,6-bisphosphate (FBP) for activation. PKR deficiency leads to hemolysis of red blood cells resulting in anemia. Activation of PKR in both sickle cell disease and beta-thalassemia patients could lead to improved red blood cell fitness and survival.The discovery of a novel series of substituted urea PKR activators, via the serendipitous identification and diligent characterization of a minor impurity in an High Throughput Screening (HTS) hit will be discussed.

Pseudohyperkalemia in pediatric patients with newly diagnosed hematological malignancies

Patients with newly diagnosed hematological malignancies often present with a considerable cellular burden, leading to complications including hyperkalemia. However, pseudohyperkalemia, arising from in vitro cell lysis, can pose challenges in clinical practice. Although pseudohyperkalemia is frequently reported in adult hematological malignancies, its occurrence in pediatric patients is underreported, and its incidence in this demographic remains unclear. We retrospectively reviewed the medical records of pediatric patients who received a new diagnosis of hematological malignancies from 2011 to 2022 at Taichung Veterans General Hospital. Hyperkalemia was defined by a serum or plasma potassium level exceeding 5.5 mEq/L. Pseudohyperkalemia was defined by 1) a potassium decrease of over 1 mEq/L in within 4 h without intervention or 2) the absence of electrocardiography changes indicative of hyperkalemia. Cases with apparent red blood cell hemolysis were excluded. A total of 157 pediatric patients with a new diagnosis of hematological malignancies were included, 14 of whom exhibited hyperkalemia. Among these 14 cases, 7 cases (4.5%) were of pseudohyperkalemia. This rate increased to 21.2% in patients with initial hyperleukocytosis. Pseudohyperkalemia was associated with a higher initial white blood cell count and lower serum sodium level. All episodes of pseudohyperkalemia occurred in the pediatric emergency department, where samples were obtained as plasma, whereas all true hyperkalemia cases were observed in the ordinary ward or intensive care unit, where samples were obtained as serum. Timely recognition of pseudohyperkalemia is crucial to avoiding unnecessary potassium-lowering interventions in pediatric patients with newly diagnosed hematological malignancies.

Analysis of non-physiological shear stress-induced red blood cell trauma across different clinical support conditions of the blood pump.

Systematic research into device-induced red blood cell (RBC) damage beyond hemolysis, including correlations between hemolysis and RBC-derived extracellular vesicles, remains limited. This study investigated non-physiological shear stress-induced RBC damage and changes in related biochemical indicators under two blood pump clinical support conditions. Pressure heads of 100 and 350mmHg, numerical simulation methods, and two in vitro loops were utilized to analyze the shear stress and changes in RBC morphology, hemolysis, biochemistry, metabolism, and oxidative stress. The blood pump created higher shear stress in the 350-mmHg condition than in the 100-mmHg condition. With prolonged blood pump operation, plasma-free hemoglobin and cholesterol increased, whereas plasma glucose and nitric oxide decreased in both loops. Notably, plasma iron and triglyceride concentrations increased only in the 350-mmHg condition. The RBC count and morphology, plasma lactic dehydrogenase, and oxidative stress across loops did not differ significantly. Plasma extracellular vesicles, including RBC-derived microparticles, increased significantly at 600min in both loops. Hemolysis correlated with plasma triglyceride, cholesterol, glucose, and nitric oxide levels. Shear stress, but not oxidative stress, was the main cause of RBC damage. Hemolysis alone inadequately reflects overall blood pump-induced RBC damage, suggesting the need for additional biomarkers for comprehensive assessments.

Ginseng total saponin improves red blood cell oxidative stress injury by regulating tyrosine phosphorylation and glycolysis in red blood cells

BackgroundOxidative stress is the main cause of many diseases, but because of its complex pathogenic factors, there is no clear method for treating it. Ginseng total saponin (GTS) an important active ingredients in Panax ginseng C.A. Mey (PG) and has potential therapeutic ability for oxidative stress due to various causes. However, the molecular mechanism of GTS in the treating oxidative stress damage in red blood cells (RBCs) is still unclear. PurposeThis study aimed to examine the protective effect of GTS on RBCs under oxidative stress damage and to determine its potential mechanism. MethodsThe oxidative stress models of rat RBCs induced by hydrogen peroxide (H2O2) and exhaustive swimming in vivo and in vitro was used. We determined the cell morphology, oxygen carrying capacity, apoptosis, antioxidant capacity, and energy metabolism of RBCs. The effect of tyrosine phosphorylation (pTyr) of Band 3 protein on RBCs glycolysis was also examined. ResultsGTS reduced the hemolysis of RBCs induced by H2O2 at the lowest concentration. Moreover, GTS effectively improved the morphology, enhanced the oxygen carrying capacity, and increased antioxidant enzyme activity, adenosine triphosphate (ATP) levels, and adenosine triphosphatase (ATPase) activity in RBCs. GTS also promoted the expression of membrane proteins in RBCs, inhibited pTyr of Band 3 protein, and further improved glycolysis, restoring the morphological structure and physiological function of RBCs. ConclusionsThis study shows, that GTS can protect RBCs from oxidative stress damage by improving RBCs morphology and physiological function. Changes in pTyr expression and its related pTyr regulatory enzymes before and after GTS treatment suggest that Band 3 protein is the main target of GTS in the treating endogenous and exogenous oxidative stress. Moreover, GTS can enhance the glycolytic ability of RBCs by inhibiting pTyr of Band 3 protein, thereby restoring the function of RBCs.

Anti-inflammatory, anti-oxidant and anti-ulcer activities of aqueous lyophilizate of Markhamia lutea (Bignoniaceae).

This work was carried out with a view to determining the antioxidant, anti-inflammatory and anti-ulcer properties of the aqueous lyophilized extract of Markhamia lutea. In vitro proteinases inhibition, albumin denaturation, hemolysis of red blood cells by heat, inhibition ofthe proton pump H+/K+ATPase, FRAP (Ferric reducing antioxidant power) and DPPH (1,1-diphenyl-2-picrylhydrazyl) assays were performed. In vivo, cold water immersion-induced ulceration and methylene blue-induced ulceration was usedto determine the anti-ulcer properties of the lyophilizate (100, 200 and 300 mg/kg). In vitro, the lyophilizate (400 μg/mL) significantly inhibited protein denaturation (66.65 %), hemolysis of red blood cells (56.54 %), proteinase activity (69.22 %); then theIC50 was 26.31 μg/mL on proton pump activity. It has alsodeveloped a strong ferric reducing antioxidant power(EC50=52.96 mmol FeSO4/g) as well as free radicals scavenging activity (EC50=22.38 μg/mL). In vivo, the aqueous lyophilizate (200 and 300 mg/kg) protected the gastric mucosa (70.68 and 79.00 % protection respectively) and reduced (p<0.05) acetylcholine, calcium and corticosterone concentrations. A decrease in malondialdehyde level, an increased glutathione level and an increased in catalase andSOD activities were recorded. In the methylene blue test, it significantly increased gastric fluid pH, while reducing gastric volume and improving hematological parameters inulcer animals. In addition, the histological sections showthat the aqueous lyophilizate of M.lutea protected the gastric mucosa from the deleterious effects of stress. The aqueous lyophilizate of M.lutea has anti-ulcer properties thanks to its anti-inflammatory, antioxidant and anti-secretory properties.

PAM-1: an antimicrobial peptide with promise against ceftazidime-avibactam resistant Escherichia coli infection.

Antibiotic misuse and overuse have led to the emergence of carbapenem-resistant bacteria. The global spread of resistance to the novel antibiotic combination ceftazidime-avibactam (CZA) is becoming a severe problem. Antimicrobial peptide PAM-1 offers a novel approach for treating infections caused by antibiotic-resistant bacteria. This study explores its antibacterial and anti-biofilm activities and mechanisms against CZA-resistant Escherichia. Coli (E. coli), evaluating its stability and biosafety as well. The broth microdilution method, growth curve analysis, crystal violet staining, scanning electron microscopy, and propidium iodide staining/N-phenyl-1-naphthylamine uptake experiments were performed to explore the antibacterial action and potential mechanism of PAM-1 against CZA-resistant E. coli. The biosafety in diverse environments of PAM-1 was evaluated by red blood cell hemolysis, and cytotoxicity tests. Its stability was further assessed under different temperatures, serum concentrations, and ionic conditions using the broth microdilution method to determine its minimum inhibitory concentration (MIC). Galleria mellonella infection model and RT-qPCR were used to investigate the in vivo antibacterial and anti-inflammatory effects. In vitro antibacterial experiments demonstrated that the MICs of PAM-1 ranged from 2 to 8 μg/mL, with its effectiveness sustained for a duration of 24 h. PAM-1 exhibited significant antibiofilm activities against CZA-resistant E. coli (p < 0.05). Furthermore, Membrane permeability test revealed that PAM-1 may exert its antibacterial effect by disrupting membrane integrity by forming transmembrane pores (p < 0.05). Red blood cell hemolysis and cytotoxicity tests revealed that PAM-1 exerts no adverse effects at experimental concentrations (p < 0.05). Moreover, stability tests revealed its effectiveness in serum and at room temperature. The Galleria mellonella infection model revealed that PAM-1 can significantly improve the survival rate of Galleria mellonella (>50%)for in vivo treatment. Lastly, RT-qPCR revealed that PAM-1 downregulates the expression of inflammatory cytokines (p < 0.05). Overall, our study findings highlight the potential of PAM-1 as a therapeutic agent for CZA-resistant E. coli infections, offering new avenues for research and alternative antimicrobial therapy strategies.

A miniaturized wash-free microfluidic assay for electrical impedance-based assessment of red blood cell-mediated microvascular occlusion

The production of HbS — an abnormal hemoglobin (Hb) — in sickle cell disease (SCD) results in poorly deformable red blood cells (RBCs) that are prone to microcapillary occlusion, causing tissue ischemia and organ damage. Novel treatments, including gene therapy, may reduce SCD morbidity, but methods to functionally evaluate RBCs remain limited. Previously, we presented the microfluidic impedance red cell assay (MIRCA) for rapid assessment of RBC deformability, employing electrical impedance-based readout to measure RBC occlusion of progressively narrowing micropillar openings. We describe herein the design, development, validation, and clinical utility of the next-generation MIRCA assay, featuring enhanced portability, rapidity, and usability. It incorporates a miniaturized impedance analyzer and features a simplified wash-free operation that yields an occlusion index (OI) within 15 min as a new metric for RBC occlusion. We show a correlation between OI and percent fetal hemoglobin (%HbF), other laboratory biomarkers of RBC hemolysis, and SCD severity. To demonstrate the assay’s versatility, we tested RBC samples from treatment-naïve SCD patients in Uganda that yielded OI levels similar to those from hydroxyurea (HU)-treated patients in the U.S., highlighting the role of %HbF in protecting against microcapillary occlusion independent of other pharmacological effects. The MIRCA assay could also identify a subset of HU-treated patients with high occlusion risks, suggesting that they may require treatment adjustments including a second-line therapy to improve their outcomes. This work demonstrates the potential of the MIRCA assay for accelerated evaluation of RBC health, function, and therapeutic effect in an ex vivo model of the microcapillary networks.

Evaluation into the Effects of Lead and Cadmium on Hemoglobin and Membrane Proteins in Human Erythrocytes

Heavy metals have negative impacts on human health including red blood cells hemolysis and increases oxidation processes, leading to many health issues. In this study, several blood samples were collected from people who work in North Oil Company, Kirkuk, Iraq. In addition to people who live, work away from oil fields as the control group. This investigation looks at the impact of various concentrations of cadmium and / or lead individually and combined (Cd:Pb) on erythrocytes hemolysis. The findings demonstrated that hemolysis produced by the (100 mM) were (72.0, 88.80, and 96.90\%) for (Cd, Pb, and Cd: Pb) respectively. Vitamins (B2 and C) were applied with different concentrations (0.1, 0.2, 0.3, 0.4, and 0.5 mol/L).to investigate the reduction of hemolysis percentage. The findings showed that both vitamins have positive effect in reducing the hemolysis percentage. Vitamin C has even more significant impact in reducing the hemolysis compared to Vitamin B2. In conclusion, cadmium and lead can cause erythrocyte hemolysis individually or combined, additionally, vitamins C and B2 were proved to in reducing hemolysis effectively.

Liposome Nanomedicine Based on Tumor Cell Lysate Mitigates the Progression of Lynch Syndrome-Associated Colon Cancer.

Treatment with immune checkpoint inhibitors (ICIs) has shown efficacy in some patients with Lynch syndrome-associated colon cancer, but some patients still do not benefit from it. In this study, we adopted a combination strategy of tumor vaccines and ICIs to maximize the benefits of immunotherapy. Here, we obtained tumor-antigen-containing cell lysate (TCL) by lysing MC38Mlh1KD cells and prepared liposome nanoparticles (Lipo-PEG) with a typical spherical morphology by thin-film hydration. Anti-PD-L1 was coupled to the liposome surface by the amidation reaction. As observed, anti-PD-L1/TCL@Lipo-PEG was not significantly toxic to mouse intestinal epithelial cells (MODE-K) in the safe concentration range and did not cause hemolysis of mouse red blood cells. In addition, anti-PD-L1/TCL@Lipo-PEG reduced immune escape from colon cancer cells (MC38Mlh1KD) by the anti-PD-L1 antibody, restored the killing function of CD8+ T cells, and targeted more tumor antigens to bone marrow-derived dendritic cells (BMDCs), which also expressed PD-L1, to stimulate BMDC antigen presentation. In syngeneic transplanted Lynch syndrome-associated colon cancer mice, the combination of anti-PD-L1 and TCL provided better cancer suppression than monoimmunotherapy, and the cancer suppression effect of anti-PD-L1/TCL@Lipo-PEG treatment was even better than that of the free drug. Meanwhile anti-PD-L1/TCL@Lipo-PEG enhanced the immunosuppressive tumor microenvironment. In vivo fluorescence imaging and H&E staining showed that the nanomedicine was mainly retained in the tumor site and had no significant toxic side effects on other major organs. The anti-PD-L1/TCL@Lipo-PEG prepared in this study has high efficacy and good biosafety in alleviating the progression of Lynch syndrome-associated colon cancer, and it is expected to be a therapeutic candidate for Lynch syndrome-associated colon cancer.

Immunogenic and toxic effects of graphene oxide nanoparticles in mouse skeletal muscles and human red blood cells

To investigate immunogenic and toxic effects of graphene oxide (GO) nanoparticles in mouse skeletal muscles and in human blood in vitro. GO nanoparticles prepared using a probe sonicator were supended in deionized H2O or PBS, and particle size and surface charge of the nanoparticles were measured with dynamic light scattering (DLS). Different concentrations (0.5, 1.0 and 2.0 mg/mL) of GO suspension or PBS were injected at multiple sites in the gastrocnemius muscle (GN) of C57BL/6 mice, and inflammatory response and immune cell infiltrations were detected with HE and immunofluorescence staining. We also examined the effects of GO nanoparticles on human red blood cell (RBC) morphology, hemolysis and blood coagulation using scanning electron microscope (SEM), spectrophotometry, and thromboelastography (TEG). GO nanoparticles suspended in PBS exhibited better colloidal dispersity, stability and surface charge effects than those in deionized H2O. In mouse GNs, injection of GO suspensions dose- and time-dependently resulted in sustained muscular inflammation and myofiber degeneration at the injection sites, which lasted till 8 weeks after the injection; immunofluorescence staining revealed obvious infiltration of monocytes, macrophages, dendritic cells and CD4+ T cells around the injection sites in mouse GNs. In human RBCs, incubation with GO suspensions at 0.2, 2.0 and 20 mg/mL, but not at 0.002 or 0.02 mg/mL, caused significant alterations of cell morphology and hemolysis. TEG analysis showed significant abnormalities of blood coagulation parameters following treatment with high concentrations of GO. GO nanoparticles can induce sustained inflammatory and immunological responses in mouse GNs and cause RBC hemolysis and blood coagulation impairment, suggesting its muscular toxicity and hematotoxicity at high concentrations.

100 What Are We Transfusing? A Prospective Trial Evaluating the Quality of Intraoperatively Salvaged Red Blood Cells in Spinal Deformity Surgery

INTRODUCTION: Intraoperative RBC salvage (aka Cell Saver) is widely employed in spinal deformity surgery. Unlike other surgical subspecialities wherein blood is immediately salvaged (with low potential RBC injury), modern approaches to spine surgery inherently result in collection of blood exposed to high-heat electrocautery, prolonged stasis, and abrasive pharmaceuticals, potentially resulting in RBC injury. However, this has not been studied in a scientific manner. METHODS: Patients undergoing spinal deformity surgery with Cell Saver were prospectively enrolled (N = 86). Comparison blood samples include baseline (arterial-line), allogenic (blood bank), and salvage (Cell Saver transfusate). Qualitative laboratory measures of RBC health and hemolysis were collected. Morphological assessment utilized Stimulated Raman Histology (SRH) and artificial intelligence based machine-learning algorithm. Functional RBC evaluation was measured via ektacytometry. RESULTS: Salvage blood differed significantly from baseline and allogenic blood, including an unpredictable range of RBC density (2.11-9.52 x 1000/uL), and significantly lower MCV (smaller RBCs) vs. baseline and allogenic samples (p = 0.015 and 4.05E-8, respectively). SRH revealed a high proportion (30.7%) of often irreversibly injured acanthocytes (shrunken and irregularly spiculated RBCs). Ektacytometry revealed that salvaged RBCs were poorly deformable and had impaired relaxation kinetics. Salvage blood samples had significantly higher intracellular components indicating active lysis: potassium (p = 0.019 and 8.2E-16), LDH (p = 3.3E-11 and 1.66E-6), and free-hemoglobin (p = 4.87E-7 and 0.001). The salvage blood mean hemolysis index (HI) was significantly higher than baseline (155x, p = 1.1E-10) and allogenic blood (7.23x, p = 2.29E-5). CONCLUSIONS: Intraoperative salvaged blood is composed of high proportions of irreversibly injured RBCs with HI even exceeding levels suitable for transfusion by US FDA and Council of Europe standards, properties that may decrease the suitability of salvaged RBCs as a blood replacement. Collection of postoperative laboratory data and perioperative outcomes are currently underway.