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The CARMEN-France registry is a prospective, multicenter registry in France including adult patients with a new diagnosis of immune thrombocytopenia or of autoimmune immune hemolytic anemia (2402 patients included in December 31, 2023). The recording of clinical, biological and treatment data allows detailed epidemiological and pharmacoepidemiological real-world studies. This review summarizes the CARMEN-France registry protocol, gives examples of studies conducted in the registry, and indicates future directions such as inclusion of patient reported outcomes, linkage with the French national health insurance database and linkage with other registries in Europe. [ABSTRACT FROM AUTHOR]

Shiga toxin-associated hemolytic uremic syndrome (STEC HUS) is the main cause of acute kidney injury in children and may be responsible for adverse outcomes despite an apparent quiescent period. To describe the medium- and long-term kidney outcomes of pediatric STEC HUS in a French region. A single-center, descriptive, retrospective study of STEC HUS cases that occurred at Besançon University Hospital between 1999 and 2017 in children up to 17 years of age was conducted. The primary study endpoint was the proportion of chronic kidney disease (CKD) cases at 5 years of follow-up. We included 98 consecutive patients. Among the 71 patients at the 5-year follow-up, we found 24 (34 %) patients with no adverse kidney outcome, 18 (25 %) with moderate adverse kidney outcome, and one (1.4 %) with severe adverse kidney outcome. Among the 96 patients at 1 year from the diagnosis, these figures were, respectively, 25 (26 %), 51 (53 %), and two (2 %); and among the 38 patients at 10 years, they were, respectively, nine (24 %), 12 (32 %), and one (3 %). The glomerular filtration rate level and oliguria–anuria beyond 8 days at baseline were significantly associated with more severe kidney outcomes at 10 years (p = 0.03 and 0.005, respectively). Two patients died during the acute phase. Overall, 33 patients (34 %) were lost to follow-up. Adverse kidney outcomes may appear many years after an episode of STEC HUS despite an apparent quiescent period. Regular long-term monitoring is required. The challenge is to reduce the proportion of patients lost to follow-up with potentially severe adverse kidney outcomes and no evaluation or treatment. [ABSTRACT FROM AUTHOR]

Alpha hemolysin (HlyA) is a hemolytic and cytotoxic protein secreted by uropathogenic strains of E. coli. The role of glycophorins (GPs) as putative receptors for HlyA binding to red blood cells (RBCs) has been debated. Experiments using anti -GPA/GPB antibodies and a GPA-specific epitope nanobody to block HlyA-GP binding on hRBCs, showed no effect on hemolytic activity. Similarly, the hemolysis induced by HlyA remained unaffected when hRBCs from a GPAnull/GPBnull variant were used. Surface Plasmon Resonance experiments revealed similar values of the dissociation constant between GPA and either HlyA, ProHlyA (inactive protoxin), HlyAΔ914-936 (mutant of HlyA lacking the binding domain to GPA) or human serum albumin, indicating that the binding between the proteins and GPA is not specific. Although far Western blot followed by mass spectroscopy analyses suggested that HlyA interacts with Band 3 and spectrins, hemolytic experiments on spectrin-depleted hRBCs and spherocytes, indicated these proteins do not mediate the hemolytic process. Our results unequivocally demonstrate that neither glycophorins, nor Band 3 and spectrins mediate the cytotoxic activity of HlyA on hRBCs, thereby challenging the HlyA-receptor hypothesis. This finding holds significant relevance for the design of anti-toxin therapeutic strategies, particularly in light of the growing antibiotic resistance exhibited by bacteria. [Display omitted] • Anti- GPA, anti GPA-GPB and nanoantibody iH4 do not affect HlyA hemolytic activity. • GPAnull/GPBnull RBCs are as sensitive to HlyA as control RBCs. • The K D values between GP proteins and HlyA, HlyA Δ914–936 and ProHlyA are similar. • Band 3 is not involved in the lytic mechanism of HlyA. • HlyA binds in the same extent to control and spectrin-depleted ghosts. [ABSTRACT FROM AUTHOR]

• Positive direct antiglobulin tests (DAT) have been reported in cases of post-artesunate delayed hemolysis (PADH) during severe malaria. • Patients with DAT positivity were not associated with PADH. • DAT does not appear to be a marker of PADH. • Overall, outcomes were favorable without corticosteroids, even in cases of PADH. Objectives: Positive direct antiglobulin tests (DATs) have been reported in cases of post-artesunate delayed hemolysis (PADH), but the causal role of auto-immune hemolysis remains unclear. We aimed to analyze a cohort of patients with PADH and DAT during severe malaria. Methods: We describe PADH and DAT results in a 7-year multi-center retrospective cohort of patients receiving artesunate for severe imported malaria. Results: Of 337 patients treated with artesunate, 46 (13.6%) had at least one DAT result within 30 days of treatment initiation, and 25/46 (54.3%) had at least one positive DAT. Among 40 patients with available data, 17 (42.5%) experienced PADH. Patient characteristics were similar for patients with a positive or negative DAT, and DAT positivity was not associated with PADH occurrence (P = 0.36). Among patients, 5/13 (38.5%) with a positive DAT after day 7 experienced PADH, compared to 10/13 (76.9%) of those with a negative DAT after day 7 (P = 0.11). Overall, 41% of patients required blood transfusions, and outcome was favorable without corticosteroids, even in cases of PADH. Conclusions : DAT does not appear to be a marker of PADH, but rather an indirect marker of an immune-mediated mechanism. DAT positivity should not lead to the administration of systemic corticosteroids during PADH. [ABSTRACT FROM AUTHOR]

A 79-year-old female with chronic kidney disease (CKD) and transthoracic aortic valve replacement presented with exertional dyspnea and was found to have hemolysis due to moderate paravalvular leak. Balloon dilatation resolved symptoms and anemia. Detecting hemolysis related to paravalvular leak is challenging due to CKD, possibly leading to bone marrow suppression. • Challenges associated with of detecting hemolytic anemia post-TAVR • Hemolytic anemia in a patient with anemia of kidney disease might not be straight-forward. • The timing to repair para-valvular leak post-TAVR needs to be further studied. [ABSTRACT FROM AUTHOR]

We aimed to verify the relationship between blood vessel shape and hemolysis risk by using a blood rheological model that reflects the physiological processes related to blood flow. Blood rheology depends on many factors including the red blood cell concentration and local shear stress, which affect the hemolysis process. We introduced a rheology model suitable for modeling hemolysis flows observed in arteries with atherosclerotic lesions in vivo based on the population balance. The model predicts the concentration of single red blood cells and the concentration and size of red blood cell agglomerates, which affect blood rheology and hemolysis. Atherosclerotic lesion geometries were obtained based on image reconstructions from tomographic projections. Computational fluid dynamics (CFD) simulation results were compared with particle image velocimetry measurements of the geometries printed with a 3D printer. Based on the CFD simulation data, a correlation function was established by combining the essential parameters of vessel shape and flow rate with the maximum shear stress, which governs the hemolysis. The chemical engineering methodology was successfully applied to the analysis of biological systems. In future, the model can be implemented in image reconstruction software using tomographic projections to quickly analyze hemolysis risk in medical practice. • Blood vessel shape and hemolysis risk are verified using a rheology model. • The introduced model predicts red blood cell concentration and agglomerate size. • Atherosclerotic lesion geometries are found using tomographic image reconstruction. • Model parameters are correlated with maximum shear stress. • The model can be used for quick hemolysis risk analysis in medical practice. [ABSTRACT FROM AUTHOR]

Sporadic mechanically-induced hemolysis associated with kinks in extracorporeal blood circuits during hemodialysis is a rare but potentially serious complication that exhibits laboratory features consistent with both in vivo and in vitro hemolysis. Misclassification of clinically significant hemolysis as in vitro can lead to inappropriate test cancellation and delayed medical interventions. Here, we report three cases of hemolysis attributed to kinked hemodialysis blood lines, which we have defined as " ex vivo " hemolysis. All three cases demonstrated an initial mixed picture of laboratory features consistent with both classifications of hemolysis. Specifically, absent features of in vivo hemolysis on blood film smear despite normal potassium led to the misclassification of these samples as in vitro hemolysis and their cancellation. A proposed mechanism for these overlapping laboratory features is the recirculation of damaged red blood cells from the kinked or pinched hemodialysis line back into the patient circulation producing an " ex vivo " hemolysis presentation. In two of the three cases, the patients developed acute pancreatitis as a result of hemolysis and required urgent medical follow up. We developed a decision pathway to help laboratories in identifying and handling these samples by recognizing that in vitro and in vivo hemolysis have overlapping laboratory features. These cases highlight the need for laboratorians and the clinical care team to be vigilant about mechanically-induced hemolysis from the extracorporeal circuit during hemodialysis. Communication is critical to identify the cause of hemolysis in these patients and prevent unnecessary delays in result reporting. [ABSTRACT FROM AUTHOR]

Sickle cell disease (SCD) is a genetic disorder that causes physical and cognitive impairment due to hemolysis, painful vaso-occlusion episodes, joint avascular necrosis, and strokes. As individuals with SCD age and develop conditions impacting their physical and cognitive function, their ability to multitask successfully and safely may decline. Cognitive-motor dual-task interference occurs when there is deterioration in one or both tasks while dual-tasking relative to single-tasking. Dual-task assessment (DTA) is a valuable measure of physical and cognitive function; however, there is limited data on DTA in adults with SCD. Is DTA a feasible and safe method of measuring physical and cognitive function in adults with SCD? What patterns of cognitive-motor interference occur in adults with SCD? We enrolled 40 adults with SCD (mean age 44 years, range 20–71) in a single-center prospective cohort study. We used usual gait speed as the measure of motor performance and verbal fluency (F, A, and S) as the measure of cognitive performance. We measured feasibility as the proportion of consented participants able to complete the DTA. We calculated the relative dual-task effect (DTE %) for each task and identified patterns of dual-task interference. Most consented participants completed the DTA (91%, 40/44) and there were no adverse events. There were 3 main dual-task interference patterns for the first trial using letter 'A′: Motor Interference (53%, n = 21), Mutual Interference (23%, n = 9), and Cognitive-Priority Tradeoff (15%, n = 6). For the second trial using letter 'S′, there were two main dual-task interference patterns: Cognitive-Priority Tradeoff (53%, n = 21) and Motor Interference (25%, n = 10). DTA was feasible and safe in adults with SCD. We identified specific patterns of cognitive-motor interference. This study supports further evaluation of DTA as a potentially useful tool to measure physical and cognitive function in ambulatory adults with SCD. • The main types of interference were motor interference, mutual interference, and cognitive-priority tradeoff (first trial) and cognitive priority tradeoff and motor interference (second trial). The change in interference patterns was mainly in participants 18–49 years old. • Age had a minor impact on performance; slightly more participants 50 or older showed cognitive or mutual interference than those 49 and younger. • The dual-task assessment is a feasible method of evaluating physical and cognitive function in ambulatory adults with SCD. [ABSTRACT FROM AUTHOR]

Quantity not sufficient (QNS) specimens with minimal blood volume for testing are common in clinical laboratories. However, there is no universal definition of minimum volume for a QNS specimen and little data is available addressing the impact of QNS / low volume specimens on turnaround time (TAT) and sample hemolysis. We compared the TAT and hemolysis index from samples ≤1.0 mL to all specimens received and quantified the number of specimens with reduced blood volume. A new QNS policy requiring ≥1.5 mL of sample in a blood tube for laboratory analysis was implemented and the results were assessed by sample hemolysis and TAT. The median laboratory TAT for samples with ≤1.0 mL of blood was 61 min (Interquartile Range, IQR: 50–82), in contrast to 28 min (26–34) for all samples. The hemolysis index for samples ≤1.0 mL was 112 (65–253) and 15 (8–29) for all samples. Requirement of a minimum volume of 1.5 mL of blood resulted in the proportion of samples with TAT ≥ 60 min to decrease from 10.4% to 4.24% in the ED, and for specimens cancelled due to hemolysis to decrease from 4.24% to 3.38%. This policy was introduced hospital wide with similar effects. Together, we correlate limited specimen volume with an increase in laboratory TAT and hemolysis. Implementation of a QNS policy of ≥1.5 mL and provider education provided a significant and durable reduction in TAT and specimen hemolysis. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a common bacterium that causes a variety of infections in humans. This microorganism produces several virulence factors, including hemolysins, which contribute to its disease-causing ability. The treatment of S. aureus infections typically involves the use of antibiotics. However, the emergence of antibiotic-resistant strains has become a major concern. Therefore, vaccination against S. aureus has gained attention as an alternative approach. Vaccination has the advantage of stimulating the immune system to produce specific antibodies that can neutralize bacteria and prevent infection. However, developing an effective vaccine against S. aureus has proven to be challenging. This study aimed to use in silico methods to design a multi-epitope vaccine against S. aureus infection based on hemolysin proteins. The designed vaccine contained four B-cell epitopes, four CTL epitopes, and four HTL epitopes, as well as the ribosomal protein L7/L12 and pan-HLA DR-binding epitope, included as adjuvants. Furthermore, the vaccine was non-allergenic and non-toxic with the potential to stimulate the TLR2-, TLR-4, and TLR-6 receptors. The predicted vaccine exhibited a high degree of antigenicity and stability, suggesting potential for further development as a viable vaccine candidate. The population coverage of the vaccine was 94.4 %, indicating potential widespread protection against S. aureus. Overall, these findings provide valuable insights into the design of an effective multi-epitope vaccine against S. aureus infection and pave the way for future experimental validations. [Display omitted] • S. aureus is an important pathogen worlwide with increasing antibiotic resistance. • Hemolysin proteins from S. aureus are highly antigenic and good candidates for a vaccine design. • Epitopes were predicted and used for the design of a multi-epitope vaccine against S. aureus using. immunoinformatic tools. • Immunological characteristics of the final multi-epitope vaccine showed promising results. [ABSTRACT FROM AUTHOR]

Ethno-pharmacological relevance : Globally, the prevalence of sickle cell disease is on the rise, with developing countries experiencing particularly alarming mortality rate compared to developed nations. The World Health Organization (WHO) and United Nations (UN) have acknowledged sickle cell disease as a significant global public health concern. Unfortunately, a cure for this condition is yet to be discovered, and existing allopathic treatments, while offering relief, come with serious side effects. In recent times, there has been a growing interest in exploring the potential of medicinal plants for treating sickle cell disease due to their content of secondary metabolites that may impact the disease's mechanisms. Cajanus cajan , a crucial grain legume in rain-fed agriculture in semi-arid tropics, has been traditionally used in folk medicine to manage various illnesses and is suggested to possess anti-sickling properties. Aim of the study : The present study investigated two varieties of C. cajan for their effectiveness in treating sickle cell beta thalassemia, a variant of sickle cell disease. The study was divided into four groups consisting of the untreated group (group 1), group treated with standard drug (group 2), group treated with white C. cajan (group 3) and group treated with brown C. cajan (group 4). The effects of the two variety of C. cajan were measured by polymerization test, reversibility test, osmotic fragility test, deoxygenation and beta globin synthesis test. The results revealed that both varieties of C. cajan demonstrated a reduction in polymerization rates, reversed sickled red blood cells, increased the oxygen affinity of Hb-S/β, elevated the Fe2+/Fe3+ ratio, and maintained the membrane stability of red blood cells. Notably, the white variety exhibited superior anti-sickling properties compared to the brown variety. This suggests that this significant leguminous crop could be utilized for the treatment and management of sickling disorders, particularly in low-income countries where conventional treatments may be financially inaccessible to patients. [Display omitted] • Two varieties of Cajanus cajan were investigated to ascertain their anti-sickling activities. • Analysis was done to measure the anti-sickling activities of the two plant varieties. • White C. cajan showed stronger anti-sickling activity compared to brown C. cajan. [ABSTRACT FROM AUTHOR]

We present here a case of documented paraprosthetic valvular leak following TAVI treated medically initially. This led to a poorly tolerated hemolytic anemia. We were able to correct this paraprosthetic valvular leak by a postdilation of the TAVI valve with a good result and uncomplicated follow-up. [ABSTRACT FROM AUTHOR]

• HPLC-DAD and HPLC-ESI-MS-MS showed that Chamaemelum nobile and Mentha pulegium are rich in phytoconstituents. • Hydroalcoholic extract of C. nobile and M. pulegium extracts showed good antioxidant activity in vitro. • Hydroalcoholic extract of C. nobile and M. pulegium significantly reduce AAPH-induced oxidative damage on human RBCs. • Hydroalcoholic extract of C. nobile and M. pulegium significantly attenuated lipidid peroxidation in human RBCs Chamaemelum nobile and Mentha pulegium are two herbs used in traditional medicine throughout the world, including Algeria. The present research aimed to study the phenolic composition of C. nobile (CN) and M. pulegium (MP) hydroalcoholic extracts, as well as their antioxidant and antihemolytic effects in AAPH-induced hemolysis on human erythrocytes. Colorimetric estimation of total polyphenols, total flavonoids, flavones and flavonols and condensed tannins showed that CN was richer in total flavonoids, flavones and flavonols and condensed tannins than MP, while MP was richer in total polyphenols. A total of 46 compounds were found using HPLC/DAD and HPLC/ESI-MS-MS analysis, of which 39 were identified. All these identified compounds are divided into five structural groups; phenolic acids, quinic acid esters, flavonoids, caffeic acid oligomers and fatty acids. For the CN, 27 compounds were identified. In contrast, for the MP, only 18 compounds were identified. Both extracts showed good antioxidant activity in vitro. The IC 50 for DPPH radical scavenging activity were 19.98 ± 0.91 µg/mL for MP and 52.77 ± 1.53 µg/mL for CN. The MP had a reducing power of 223.74 ± 1.08 mg ascorbic acid equivalent/g extract, while CN's was 130.65 ± 1.69 mg Asc E/g. Both extracts were also significantly effective and in a dose-dependent manner in reducing AAPH-induced oxidative damage on erythrocyte membranes with IC50s of 127.48 ± 3.14 µg/mL for CN and 129.52 ± 2.15 µg/mL for MP. Lipid peroxidation induced by AAPH and estimated by MDA levels revealed that both extracts significantly reduced MDA levels at doses of 200 and 300 µg/mL, particularly MP. Our findings provide evidence that C. nobile and M. pulegium have a considerable protective effect on the erythrocyte membrane against free radicals generated by AAPH. [ABSTRACT FROM AUTHOR]

• Hydroxocobalamin infusion can mimic hemolysis in plasma samples. • Hydroxocobalamin does not interfere with 3,3′,5,5′-tetramethylbenzidine based methodologies for plasma hemoglobin measurement. • Hydroxocobalamin may continue to be present for >5 days following infusion. Hemolysis is one of the most common preanalytical concerns in the clinical laboratory. Hydroxocobalamin administration causes red pigmentation of plasma that may mimic hemolysis and may interfere with chemistry assays. A male patient in his sixties was placed on extracorporeal membrane oxygenation (ECMO) as a bridge to transplantation. Daily plasma free hemoglobin measurements were ordered to monitor for adverse ECMO events. An intensely red plasma specimen was inconsistent with modestly elevated hemoglobin levels and became pink on dilution. Follow-up with providers indicated that the red plasma could be attributed to hydroxocobalamin administration. Performance of scanning spectrophotometry and assessment of a sample spiked with hydroxocobalamin indicated that the red colored hydroxocobalamin did not interfere with our 3,3′,5,5′-tetramethylbenzidine based methodology for free plasma hemoglobin measurement. It is important for the laboratory professionals to be aware of the possibility of interference in hemoglobin assays due to hydroxocobalamin. [ABSTRACT FROM AUTHOR]

• Patients with wasp stings experience a significant reduction in blood lipid levels. • Hemolysis and inflammatory responses might lead to hypolipidemia. • Early blood lipid levels have a relationship with the clinical severity of wasp stings. • Lipid metabolism might be a new target for future treatment of wasp stings. Wasp stings are a serious problem worldwide, and patients in severe cases may experience multi-organ failure. However, the mechanism of hypolipidemia in patients with wasp stings is unknown. To investigate the relationship between early changes in lipid levels and clinical severity and the possible underlying mechanisms. A retrospective analysis of 212 patients (mild: 77; moderate: 50; severe: 85) with wasp stings was conducted. Clinical data, including lipid test results within 24 h of admission, were analysed. A total of 1060 healthy age- and gender-matched controls were used. Patients with wasp stings had lower lipid levels than healthy controls (P <0.01). Lipid levels decreased with disease severity, except for triglycerides (P <0.05). The number of stings, degree of organ failure, need for mechanical ventilation and extracorporeal blood purification, and mortality were higher in the severe group than in the mild and moderate groups (P <0.01). A decrease in lipid levels was accompanied by an increase in inflammatory indicators. In the severe group, a reduction in lipid levels was associated with ventilator application and blood purification, independent of survival status. Patients with wasp stings experience a reduction in lipid levels, which is related to the severity of clinical manifestations. Early lipid levels may serve as a simple indicator for the severity of wasp stings, and targeting lipid metabolism may be a novel treatment. [ABSTRACT FROM AUTHOR]

• FDA centrifugal blood pump benchmark is investigated under non-deterministic conditions. • Stochastic condition can explain a part of discrepancy between the numerical and experimental results. • Uncertainties lead to remarkable variations in the pump hemodynamics and performance characteristics. • The hemolysis index is profoundly affected by the uncertainties in model parameters. • Impeller diameter and rotational speed are the most important parameters on the hydrodynamic characteristics. The present study is set out to systematically investigate the combined impact of operational, geometrical, and model uncertainties on the hemodynamics and performance characteristics in the U.S. Food and Drug Administration (FDA) benchmark centrifugal blood pump. Non-intrusive Polynomial Chaos Expansion (NIPCE) has been utilized to propagate the uncertainty of 12 random input variables in the flow field and the performance characteristics of the blood pump at three working conditions. The global sensitivity of the Quantities of Interest (QoI) to the uncertain input parameters was measured through the Sobol' indices. The Multiple Reference Frames (MRF) approach and the SST k − ω turbulence model were employed to conduct the 3D CFD computations of the pump. In addition, we quantified the device-related hemolysis using the semi-empirical power-law model. The stochastic CFD results of the pump velocity field and hydraulic performance parameters showed a satisfactory agreement with measurements. Statistical analysis indicated that the effect of operational and geometrical uncertainties on the velocity field of the blood pump chiefly emerges at the outlet diffuser region, more specifically along the center-line of the fluid jet formed inside the diffuser. This study has also clarified that the uncertainties in the flow field and the hydraulic performance of the blood pump are mainly due to the variations of impeller diameter, rotational speed, radial clearance, and flow rate. By contrast, the hemolysis index is profoundly affected by the model parameters. Additionally, higher robustness against uncertainties was observed for hydraulic efficiency compared to the pump head. Eventually, it was shown that the maximum value of the distribution function of the relative hemolysis index lies within the bounds of the measurements. [ABSTRACT FROM AUTHOR]

• Latent hemolytic patients showed negative results for usual hemolysis indicators. • Erythrocyte creatine (EC) is known to reflect the mean erythrocyte age (M RBC). • Recently, we reported a formula for obtaining M RBC based on EC. • M RBC values based on EC were lower in two patients with latent hemolysis. • Measurement of EC might be useful for the diagnosis of latent hemolysis. Whereas HbA1c values are low relative to glycemia in patients with hemolytic anemia, including compensatory anemia, low HbA1c levels along with negative results for conventional hemolysis indicators have been reported in patients with latent hemolysis. Conversely, glycated albumin (GA) is a glycemic control indicator unaffected by hemolysis. Erythrocyte creatine (EC) is a hemolysis indicator that reflects the mean age of red blood cells (M RBC). We recently reported a formula for obtaining M RBC based on EC. The present study examined the usefulness of EC measurements and M RBC calculated with EC for diagnosing latent hemolysis. Two patients with latent hemolysis and low HbA1c values relative to glycemia were investigated, while controls comprised 214 patients (including patients with hemolysis and/or type 2 diabetes mellitus). HbA1c was expressed in International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) units (iA1c). GA/iA1c ratios, reticulocyte counts, EC, and M RBC in patients with latent hemolysis were compared to non-hemolysis, compensatory hemolysis, and hemolytic anemia patients. Both reticulocyte counts and haptoglobin levels were within reference ranges in patients with latent hemolysis. GA/iA1c ratios and EC were higher than reference values in patients with latent hemolysis, and M RBC values were 41.6 and 48.4 days, respectively, shorter than the reference range (49.1–66.8 days). EC measurement and M RBC values calculated on the basis of EC might be useful for diagnosing latent hemolysis. [ABSTRACT FROM AUTHOR]

Absolute coronary flow can be measured by intracoronary continuous thermodilution of saline through a dedicated infusion catheter (RayFlow®). A saline infusion rate at 15–20 mL/min induces an immediate, steady-state, maximal microvascular vasodilation. The mechanism of this hyperemic response remains unclear. We aimed to test whether local hemolysis is a potential mechanism of saline-induced coronary hyperemia. Twelve patients undergoing left and right catheterization were included. The left coronary artery and the coronary sinus were selectively cannulated. Absolute resting and hyperemic coronary flow were measured by continuous intracoronary thermodilution. Arterial and venous samples were collected from the coronary artery and the coronary sinus in five phases: baseline (BL); resting flow measurement (Rest , saline infusion at 10 mL/min); hyperemia (Hyperemia, saline infusion at 20 mL/min); post-hyperemia (Post-Hyperemia, 2 min after the cessation of saline infusion); and control phase (Control , during infusion of saline through the guide catheter at 30 mL/min). Hemolysis was visually detected only in the centrifugated venous blood samples collected during the Hyperemia phase. As compared to Rest , during Hyperemia both LDH (131.50 ± 21.89 U/dL [ Rest ] and 258.33 ± 57.40 U/dL [ Hyperemia ], p < 0.001) and plasma free hemoglobin (PFHb, 4.92 ± 3.82 mg/dL [ Rest ] and 108.42 ± 46.58 mg/dL [ Hyperemia ], p < 0.001) significantly increased in the coronary sinus. The percentage of hemolysis was significantly higher during the Hyperemia phase (0.04 ± 0.02% [ Rest ] vs 0.89 ± 0.34% [ Hyperemia ], p < 0.001). Saline-induced hyperemia through a dedicated intracoronary infusion catheter is associated with hemolysis. Vasodilatory compounds released locally, like ATP, are likely ultimately responsible for localized microvascular vasodilation. [Display omitted] • Absolute coronary flow and resistance can be measured with continuous intracoronary thermodilution of saline through the RayFlow catheter. • The mechanisms underlying saline-induced hyperemia are still unknown. Local hemolysis might be involved. • Serum markers of hemolysis were measured from blood samples collected from the coronary artery and the coronary across different phases of the study. • There was a significant increase in LDH, plasma free Hb and % of hemolysis during hyperemia as compared to baseline and resting conditions. • Saline-induced coronary hyperemia is mediated by intravascular hemolysis, likely through the local release of vasodilatory compounds (i.e. ATP, NO). [ABSTRACT FROM AUTHOR]

Hemolysis is the most prevalent pre-analytical interfering factor and a major source of error in laboratory analysis. The examination of samples post-centrifugation can provide valuable information regarding pre-analytical interferences. In this unusual case, a patient's plasma specimen was cherry-red after centrifugation, which is most usually indicative of hemolysis. However, subsequent investigations ruled out common hemolysis causes. We eventually determined that the patient's cherry-red plasma was most likely caused by other factors in the patient's medical history, including cancer treatment with PV-10 (rose bengal disodium 10%). We then conducted an interference study to comprehensively assess the effects of PV-10 on various biochemical tests, especially liver function tests and bilirubin levels. The findings indicate that PV-10 has varying effects on different biochemical assays and test results should be examined individually. This report underlines the need for awareness of potential drug interference on laboratory tests for better result interpretation and making clinical decisions. [ABSTRACT FROM AUTHOR]

Intravascular hemolysis is associated with massive release of hemoglobin and consequently labile heme into the blood, resulting in prothrombotic and proinflammatory events in patients. Though heme is well-known to participate in these adverse effects, it is not monitored. Instead, haptoglobin and hemoglobin serve as clinical biomarkers. The quantification of labile heme together with hemoglobin, however, should be considered in clinical diagnosis as well, to obtain a complete picture of the hemolytic state in patients. So far, quantification techniques for labile heme were not yet systematically analyzed and compared for their clinical application potential, especially in the presence of hemoglobin. Two commercial assays (Heme Assay Kit®, Hemin Assay Kit®) and five common approaches (pyridine hemochromogen assay, apo-horseradish peroxidase-based assay, UV/Vis spectroscopy, HPLC, mass spectrometry) were analyzed concerning their linearity, accuracy, and precision, as well as their ability to distinguish between hemoglobin-bound heme and labile heme. Further, techniques for the quantification of hemoglobin (Harboe method, SLS method, Hemastix®) were included to study their selectivity for hemoglobin and potential interference by the presence of labile heme. Both, indirect and direct approaches were suitable for the determination of a wide concentration of heme (∼0.02–45 μM) and hemoglobin (∼0.002–17 μM). A clear distinction between hemoglobin-bound heme and labile heme with one method was not possible. Thus, a novel combined approach is presented and applied to human and porcine plasma samples for the determination of hemoglobin and labile heme. Our results demonstrate the need to develop improved techniques to differentiate labile and protein-bound heme for early detection of intravascular hemolysis. Here, we present a novel strategy by combining two spectroscopic methods, which is most reliable as an easy-to-use tool for the determination of hemoglobin and heme levels in plasma samples for the diagnosis of intravascular hemolysis and in basic biomedical research. [Display omitted] • Heme initiates prothrombotic and proinflammatory conditions in hemolytic events. • Heme and hemoglobin detection methods are evaluated by bioanalytical test criteria. • Available methods cannot distinguish between hemoglobin-bound heme and labile heme. • A strategy for heme detection in blood plasma samples is provided. [ABSTRACT FROM AUTHOR]

Cytotoxicity studies determining hemolytic properties of antimicrobial peptides or other drugs are an important step in the development of novel therapeutics for clinical use. Hemolysis is an affordable, accessible, and rapid method for initial assessment of cellular toxicity for all drugs under development. However, variability in species of red blood cells and protocols used may result in significant differences in results. AMPs generally possess higher selectivity for bacterial cells but can have toxicity against host cells at high concentrations. Knowing the hemolytic activity of the peptides we are developing contributes to our understanding of their potential toxicity. Computational approaches for predicting hemolytic activity of AMPs exist and were tested head-to-head with our experimental results. Starting with an observation of high hemolytic activity of LL-37 peptide against human red blood cells that were collected in EDTA, we explored alternative approaches to develop a more robust, accurate and simple hemolysis assay using defibrinated human blood. We found significant differences between the sensitivity of defibrinated red blood cells and EDTA treated red blood cells. Accurately determining the hemolytic activity using human red blood cells will allow for a more robust calculation of the therapeutic index of our potential antimicrobial compounds, a critical measure in their pre-clinical development. We introduce a standardized, more accurate protocol for assessing hemolytic activity using defibrinated human red blood cells. This approach, facilitated by the increased commercial availability of de-identified human blood and defibrination methods, offers a robust tool for evaluating toxicity of emerging drug compounds, especially AMPs. [Display omitted] [ABSTRACT FROM AUTHOR]

Hemocompatibility evaluation is an important step in nanotoxicological studies. It is generally accepted that nanomaterials promote lysis of erythrocytes, blood clotting, alter phagocytosis, and upregulate pro-inflammatory cytokines. However, there are no standardized guidelines for testing nanomaterials hemocompatibility despite the fact that nanomaterials enter the bloodstream and interact with blood cells. In this review, the current knowledge on the ability of nanomaterials to induce distinct cell death modalities of erythrocytes is highlighted primarily focusing on hemolysis and eryptosis. This review aims to summarize the molecular mechanisms underlying erythrotoxicity of nanomaterials and critically compare the sensitivity and efficiency of hemolysis or eryptosis assays for nanomaterials blood compatibility testing. The list of eryptosis-inducing nanomaterials is growing, but it is still difficult to generalize how physico-chemical properties of nanoparticles affect eryptosis degree and molecular mechanisms involved. Thus, another aim of this review is to raise the awareness of eryptosis as a nanotoxicological tool to encourage the corresponding studies. It is worthwhile to consider adding eryptosis to in vitro nanomaterials hemocompatibility testing protocols and guidelines. • Diverse nanomaterials induce eryptosis, a regulated apoptosis-like cell death mode of mature erythrocytes. • Eryptosis detection is an efficient tool in nanotoxicological studies. • Eryptosis assays can supplement hemolysis assays in studies investigating hemocompatibility of nanomaterials. [ABSTRACT FROM AUTHOR]

• The Alinity c HI allows the quantitative determination of plasma free hemoglobin. • The method is rapid and cost-effective with a measuring range from 80 to 7250 mg/L. • Bilirubin up to 710 mg/L and Intralipid® up to 5580 mg/L did not affect the measurement. • The method can be used for STAT analysis in a routine laboratory. Quantitative measurement of plasma free hemoglobin (fHb) concentrations is essential for monitoring pediatric ECMO patients, since hemolysis has a great impact on the patient's clinical outcome. The aim of this study was to validate the hemolysis index (HI) assay on Abbott's Alinity c system as a quantitative method to measure fHb. The performance of the HI assay, based on an automated spectrophotometric method recording the absorption at four different wavelength pairs, was evaluated using the 20 × 2 × 2 design according to the CLSI-EP05-A3 guidelines. LLOQ and LLOD were calculated according to CLSI-EP17 guidelines with CVs set to 10% and 20%, respectively. Furthermore, the method was tested for interferences with bilirubin and Intralipid®. Linearity was ensured over an analytical measurement range of 30–7250 mg/L and the calculated LLOQ and LLOD were 80 mg/L and 50 mg/L, respectively. Intra-run and total imprecisions ranged from 0.9–3.4% and 1.0–3.4%, respectively. The HI assay correlated well with the Harboe method (HI (mg/L) = 0.998 * fHb (mg/L) + 28 mg/L, R = 0.998, n = 50) and interference testing showed no impact of bilirubin and Intralipid® up to 709 mg/L and 5580 mg/L, respectively. The HI assay on Abbott's Alinity c system allows a precise and accurate determination of fHb concentrations with no significant interferences in a simple, rapid and cost-effective way. [ABSTRACT FROM AUTHOR]

In response to increasing emergency department presentations and wait times in Australia, multiple strategies and models of care have been implemented with varying results. One effective strategy has been the implementation of pre-hospital blood collection by paramedics when they insert an intravenous cannula. This research aims to determine the efficiency of and barriers to wider implementation of a pre-hospital blood collection trial in a regional context. In particular, to evaluate the impact of the pre-hospital blood collection on time to pathology results and error rates, and paramedic opinion. This retrospective controlled cohort study was conducted over 12 months from August 2018. Emergency and pathology data were used to determine the haemolysis and error rates, as well as the time to result availability of pre-hospital blood collection samples compared to in hospital samples arrived by ambulance. To determine the facilitators and barriers to wider implementation a survey of 48 paramedics was conducted following completion of the 12-month trial. The survey was informed by the Theoretical Domains Framework, a behavior change theory associated with improved uptake when applied. Overall 237 samples were collected. There was a 65% (51 min) reduction in time taken for samples to be received at pathology and a 38% (50 min) improvement in time taken for results to return from pathology for patients arrived by ambulance. There were no labelling errors in the pre-hospital blood collection group or change in haemolysis rates. The majority (79%) of paramedics who completed the survey were optimistic about the protocol improving patient outcomes and 89% regarded the change in practice as acceptable. Three main themes emerged: 1. Training, environmental challenges and adequate equipment; 2. increased efficiency and improved patient care and 3. Prerequisites to implementing a new practice. Integration of Quantitative and Qualitative data resulted in 10 key influencers of behavior that need to be addressed in any future implementation. The introduction of pre-hospital phlebotomy reduced the time to blood results availability by 38% and resulted in fewer labelling errors. Wider implementation is supported by paramedics, but more training is required. [ABSTRACT FROM AUTHOR]

• A new experiment was used to investigate potassium correction in hemolytic samples. • A hemolytic index restricted correction procedure was designed using 20 samples. • Validation was performed in an independent validation cohort. • Correction within TEa was possible for 70/70 validation samples (100% concordance) • Potassium correction is feasible and increased the accuracy of potassium results. For several decades, there has been an ongoing debate about the appropriateness and reliability of correcting potassium concentration results for hemolyzed samples. As part of implementing a new Roche Cobas Pro analyzer system the possibility of correcting potassium results in hemolytic samples using a new, thorough experimental design, was investigated. The relationship between hemolytic index (HI) and increases in potassium concentration was studied by performing a linear regression on hemolysate dilution series (HI 0–160 mg/dL) from 20 left-over patient samples. The obtained correction procedure was validated using another 20 left-over patient samples. Corrections were accepted according to a correction concordance of 100% within the total allowable error criterion of 4.85%. The obtained reporting procedure was: HI 0–17 quantitative potassium reporting, HI 18–100 correct potassium for HI, and report as text including a disclaimer for in vivo hemolysis; samples were rejected for HI > 100. In the validation cohort, 70/70 samples eligible for correction were within the TEa criterion. The maximum negative and positive errors were −2.8% and 2.9%, respectively. Correcting potassium concentration results in a designated HI range is feasible and increases the accuracy the potassium results in samples with mild in vitro hemolysis. [ABSTRACT FROM AUTHOR]

To assess the impact of hemolysis on laboratory results under local conditions and to verify the hemolysis index cut-off for potassium using real-world data. The statistical bootstrapping method was performed on 54,125 samples collected at the University Hospital of Örebro (USÖ). The results were compared to a method based on stratification of samples according to hemolysis level, and on paired difference testing. Setting the acceptable allowable limit of error to 10%, the three assessed strategies yielded comparable results with respect to the impact of haemolytic interference on test results for potassium. The suggested cut-offs were 111 mg Hb/dL for the bootstrapping method, between 125–150 mg Hb/dL for the method based on stratification, and around 150 mg/dL for the paired difference testing strategy. The impact of hemolysis on potassium measurement is likely different between primary care patients and inpatients. Using the effect of hemolysis on potassium measurement as a model, a novel approach towards finding clinically acceptable limits for analytical interference is presented, that relies on the bootstrapping method and on actual patient data from routine laboratory operation, hence incorporating local population characteristics, equipment and instrumental settings. [ABSTRACT FROM AUTHOR]

Sanguinarine (SGN) is a benzophenathridine alkaloid extracted from Sanguinaria canadensis plant. SGN is incriminated in epidemic dropsy (ED) characterized by multiple-organ failure and anemia. Nevertheless, how SGN leads to anemia of ED remains poorly understood. This study was thus initiated to investigate the interaction of SGN with human red blood cells (RBCs) and to delineate associated molecular mechanisms. Heparin- and EDTA-anticoagulated blood was collected from healthy participants and whole blood was analyzed for a complete blood count, while isolated RBCs were examined for hemolytic and eryptotic markers following exposure to 1–100 μM SGN for 24 h at 37 °C. Calcium was measured by Fluo4/AM, hemolysis by hemoglobin leakage, membrane scrambling by Annexin V-FITC, cell size by forward scatter (FSC), cell granularity by side scatter (SSC), and oxidative stress by H 2 DCFDA. SGN led to increased Fluo4 fluorescence and dose-dependent hemolysis which was not ameliorated by exclusion of extracellular Ca2+ but was nevertheless sensitive to hyperosmotic conditions and to the presence of aspirin. SGN also caused significant increase in Annexin V-positive cells, decreased FSC and SSC values, and elevated DCF fluorescence. Moreover, significantly reduced lymphocyte and basophil percentages along with selective toxicity to platelets was noted. Collectively, SGN possesses sucrose- and cyclooxygenase-sensitive hemolytic potential and elicits eryptosis characterized by Ca2+ accumulation, phosphatidylserine externalization, morphological alterations including cell shrinkage and loss of granularity, and oxidative stress. In conclusion, this report reveals a novel activity of SGN against human RBCs and informs prospective policies in ED prevention and management. • Sanguinarine chloride induces sucrose- and cyclooxygenase-sensitive hemolysis dose-responsively in human red blood cells. • Sanguinarine chloride elicits eryptosis with phosphatidylserine externalization, calcium overload, and oxidative stress. • Sanguinarine chloride is selectively cytotoxic to lymphocytes and platelets in a whole blood ex vivo system. [ABSTRACT FROM AUTHOR]

In this work, for the first time, zinc substituted baghdadite (Ca 3-x Zn x ZrSi 2 O 9 ; x = 0, 0.1, 0.25 and, 0.5) ceramic was successfully prepared. A solid-state ceramic synthesis route was utilized to prepare a composite for biomedical applications. The incorporation of zinc oxide reduces the synthesis temperature. The properties of composites were presented in terms of Zinc oxide amount. In-vitro bioactivity and degradation results indicated lower degradation in comparison with pure baghdadite. The hemolysis rate was observed < 5 %. Spreading of MG-63 cells in a large number was found on the baghdadite in comparison with 0.5 mol % zinc oxide containing composite. The antibacterial activity of composites analyzed over E. Coli and S. Aureus bacterial strain using MTT assay. Results showed that anti-bacterial ability was more in zinc oxide containing composite and found more active towards E. Coli. However, the antibacterial activity was found less in baghdadite ceramic. Therefore considering the improved antibacterial activities, and controlled degradation, baghdadite/zinc oxide composite can be highly considered as a promising material for biomedical application. • Baghdadite/zinc oxide composite is prepared by the solid-state method. • The incorporation of zinc oxide lowers the processing temperature. • Zinc oxide in baghdadite reduces the degradation rate and improves the antibacterial activity. • Composites are hemolytic. [ABSTRACT FROM AUTHOR]

The high burden of disease and the long-lasting sequelae following Streptococcus pyogenes (Strep A) infections make the development of an effective vaccine a global health priority. Streptolysin O (SLO), is a key toxin in the complex pathogenesis of Strep A infection. Antibodies are elicited against SLO after natural exposure and represent a key target for vaccine-induced immunity. Here we present the setup and characterization of a hemolysis assay to measure functionality of anti-SLO antibodies in human sera. Assay specificity, precision, linearity, reproducibility, and repeatability were determined. The assay was demonstrated to be highly sensitive, specific, reproducible, linear and performed well in assessing functionality of anti-SLO antibodies induced by exposed individuals. Moreover, different sources of critical reagents, in particular red- blood cells, have been compared and had minimal impact on assay performance. The assay presented here has throughput suitable for evaluating sera in vaccine clinical trials and sero-epidemiological studies to gain further insights into the functionality of infection- and vaccine-induced antibodies. [ABSTRACT FROM AUTHOR]

Mechanical intravascular hemolysis is frequently observed following procedures on heart valves and uncommonly observed in native valvular disease. In most cases, its severity is mild. Nevertheless, it can be clinically significant and even life threatening, requiring multiple blood transfusions and renal replacement therapy. This paper reviews the current knowledge on mechanical intravascular hemolysis in valvular disease, before and after correction, focusing on pathophysiology, approach to diagnosis, and impact of other hematological conditions on the resultant anemia. The importance of a multidisciplinary management is underscored. Laboratory data are provided about subclinical hemolysis that is commonly observed following the implantation of surgical and transcatheter valve prostheses and devices. Finally, clinical scenarios are reviewed and current medical and surgical treatments are discussed, including alternative options for inoperable patients. [ABSTRACT FROM AUTHOR]

During red blood cell (RBC) lysis hemoglobin and heme leak out of the cells and cause damage to the endothelium and nearby tissue. Protective mechanisms exist; however, these systems are not sufficient in diseases with increased extravascular hemolysis e.g. hemolytic anemia. α 1 -microglobulin (A1M) is a ubiquitous reductase and radical- and heme-binding protein with antioxidation properties. Although present in the circulation in micromolar concentrations, its function in blood is unclear. Here, we show that A1M provides RBC stability. A1M−/− mice display abnormal RBC morphology, reminiscent of macrocytic anemia conditions, i.e. fewer, larger and more heterogeneous cells. Recombinant human A1M (rA1M) reduced in vitro hemolysis of murine RBC against spontaneous, osmotic and heme-induced stress. Moreover, A1M is taken up by human RBCs both in vitro and in vivo. Similarly, rA1M also protected human RBCs against in vitro spontaneous, osmotic, heme- and radical-induced hemolysis as shown by significantly reduced leakage of hemoglobin and LDH. Addition of rA1M resulted in decreased hemolysis compared to addition of the heme-binding protein hemopexin and the radical-scavenging and reducing agents ascorbic acid and Trolox (vitamin E). Furthermore, rA1M significantly reduced spontaneous and heme-induced fetal RBC cell death. Addition of A1M to human whole blood resulted in a significant reduction of hemolysis, whereas removal of A1M from whole blood resulted in increased hemolysis. We conclude that A1M has a protective function in reducing hemolysis which is neither specific to the origin of hemolytic insult, nor species specific. Spontaneous, osmotic, radical- and heme-induced stress (1) all lead to hemolysis, which is further accelerated by a feed-forward loop (2) caused by leakage of hemoglobin, free heme, ROS and iron (III/IV) ions. The reductase and heme- and radical-binding protein A1M inhibits hemolysis by interfering with both steps. Image 1 • A1M-knockout mice have a steady state macrocytic anemia phenotype. • A1M reduces RBC lysis from spontaneous, osmotic, heme- and radical induced stress. • A1M reduces hemolysis in three different RBC sources: adult, fetal and murine. • A1M is internalized by RBCs both in vivo and in vitro. [ABSTRACT FROM AUTHOR]

The polysaccharides from blackcurrant (Ribes nigrum L.) fruits were degraded by ultrasonic irradiation. Results showed that viscosity-average molecular weight decreased with increasing ultrasonic time or power. The degradation was fitted to the second-order kinetics model and midpoint chain scission model. Gas chromatographic analysis demonstrated that the native polysaccharide and three degraded polysaccharides were composed of the same monosaccharides but in different ratios. Fourier transform infrared and nuclear magnetic resonance spectroscopic analyses revealed the presence of α -, β -pyranose rings and the same six sugar residues in the four blackcurrant polysaccharides. Compared to the native polysaccharide, three degraded polysaccharides displayed better rheological properties and stronger protective effects against erythrocyte hemolysis. Collectively, the results support the potential utility of blackcurrant polysaccharides as natural antioxidants. • Kinetics and chain scission models of polysaccharides degradation were established. • Three degraded polysaccharides displayed excellent rheological behaviors. • Physicochemical properties and structure of polysaccharides were analysed. • Degraded polysaccharides possessed better protection effects on erythrocyte hemolysis. [ABSTRACT FROM AUTHOR]

Leukocidin ED (LukED) is a pore-forming toxin produced by Staphylococcus aureus, which lyses host cells and promotes virulence of the bacteria. LukED enables S. aureus to acquire iron by lysing erythrocytes, which depends on targeting the host receptor Duffy antigen receptor for chemokines (DARC). The toxin also targets DARC on the endothelium, contributing to the lethality observed during bloodstream infection in mice. LukED is comprised of two monomers: LukE and LukD. LukE binds to DARC and facilitates hemolysis, but the closely related Panton-Valentine leukocidin S (LukS-PV) does not bind to DARC and is not hemolytic. The interaction of LukE with DARC and the role this plays in hemolysis are incompletely characterized. To determine the domain(s) of LukE that are critical for DARC binding, we studied the hemolytic function of LukE-LukS-PV chimeras, in which areas of sequence divergence (divergence regions, or DRs) were swapped between the toxins. We found that two regions of LukE's rim domain contribute to hemolysis, namely residues 57-75 (DR1) and residues 182-196 (DR4). Interestingly, LukE DR1 is sufficient to render LukS-PV capable of DARC binding and hemolysis. Further, LukE, by binding DARC through DR1, promotes the recruitment of LukD to erythrocytes, likely by facilitating LukED oligomer formation. Finally, we show that LukE targets murine Darc through DR1 in vivo to cause host lethality. These findings expand our biochemical understanding of the LukE-DARC interaction and the role that this toxin-receptor pair plays in S. aureus pathophysiology. [ABSTRACT FROM AUTHOR]

• The impact of hemolysis and lipemia was analyzed on a Roche cobas® c701 system. • Assessment of hemolysis and lipemia interferences is clinically important. • Native analyte level may influence the degree of acceptable endogenous interferents. • Specimen acceptability thresholds may differ in practice compared to vendor's claim. A key component of laboratory medicine is the evaluation of specimen suitability for downstream analytical testing. Accurate identification and characterization of the impact of interferents on clinical chemistry analytes is important for patient care. To empirically assess the influence of hemolysis and lipemia on clinical chemistry tests analyzed on a Roche cobas® c701 system, we evaluated serum pools spiked with increasing concentrations of hemolysate and Intralipid®. Using an interferent acceptance threshold of within ± 10% of the non-hemolyzed or non-lipemic results, 31 routine chemistry analytes were evaluated. The majority of analytes were determined to have the same or very similar acceptability thresholds as those listed in the vendor package insert. However, several analytes resulted in new thresholds that deviated from manufacturer recommendations (9 higher and 2 lower for lipemia, 7 higher and 6 lower for hemolysis). Samples with high enzyme activities (LDH, ALT, AST, ALP, and CK) were observed to tolerate higher levels of hemolysis, and tiered hemolysis thresholds were established for these enzymes. Independent evaluation of indices is recommended to enable thoughtful implementation of specimen quality criteria and to provide guidance to laboratorians and providers on the nature of these interferences. [ABSTRACT FROM AUTHOR]

• We found that the wavelength used for hemolysis index measurement should be 571 nm. • Hemolysis index measured at or near 571 nm is not influenced by conjugated bilirubin, free bilirubin or turbidity. • We developed a method to produce artificially hemolyzed specimens by using steel beads. Hemolysis is a common problem in the handling of serum specimens. The hemolysis index (HI) provides a warning of hemolysis in auto-analyzers. However, HI has not been standardized, and each laboratory's original method is applied. Especially, the wavelength used for HI measurement is different in each laboratory. Thus, we investigated the warning ability of HI at various wavelengths. We selected 4 wavelength types, and each HI was measured and calculated (410 nm/HI-1, 451 nm/HI-2, 545 nm/HI-3, and 571 nm/HI-4). To compare the 4 HI types, we investigated the influence of 3 interference components using artificially hemolyzed specimens (AHSs). We also investigated both the relationship between HI and hemoglobin concentration (Hb) and that between HI and 31 biochemical test values in AHSs. In the interference assessment, only HI-4 showed no influence on the 3 interference components. The correlation between Hb and HI-4 was very strong (r S = 0.9987). A 1-unit increase in HI-4 corresponded to a 14.8-mg/dL increase in Hb. We found the best wavelength for HI to be at or near 571 nm. [ABSTRACT FROM AUTHOR]

• There are some clues that antioxidants reduce the process of hemolysis. • Omega-3 fatty acids seem to be able to reduce hemolysis in sickle cell disease. • Efficacy of antioxidants in reducing hemolysis should be more explored. Hemolysis is one of the main pathophysiological characteristics of sickle cell disease (SCD) and might cause or could be the result of oxidative stress. Antioxidants are studied in SCD due to their potential to ensure redox balance and minimize deleterious effects on erythrocyte membranes. The objective of this systematic review was to evaluate the efficacy of antioxidant nutrient supplementation on reducing hemolysis in SCD patients through randomized clinical trials. We conducted our study according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses and the Cochrane Handbook for Systematic Reviews of Interventions investigating whether antioxidants could improve the hemolytic status of SCD patients. This study included 587 articles published until April 2020. We reduced this pool to 12 articles by excluding duplicates, reviews, comments, and studies with non-human subjects. Omega-3 fatty acids, vitamin A, and zinc were the antioxidants that reportedly improved the indirect hemolysis parameters such as hemoglobin, hematocrit, mean corpuscular volume, or red blood cells. High-dose vitamin C and E supplementation worsened hemolysis, causing increased reticulocytes, lactate dehydrogenase, indirect bilirubin, and haptoglobin. More intervention studies especially high-quality controlled randomized clinical trials are needed to investigate the effects of antioxidant nutrients in reducing hemolysis in SCD. [ABSTRACT FROM AUTHOR]