Frequent detection of functional hyposplenism via assessment of pitted erythrocytes in patients with advanced liver cirrhosis.

BACKGROUND: Asplenia or functional hyposplenism are risk factors for severe infections, and vaccinations against encapsulated bacteria are advised. There are only limited data regarding the spleen function of cirrhotic patients. METHODS: We evaluated spleen function in patients with liver cirrhosis, who were prospectively enrolled in this study. Spleen function was evaluated by the measurement of pitted erythrocytes. Functional hyposplenism was defined as a percentage of PE of >15%. RESULTS: 117 patients, mean age 58.4 years and 61.5% (n = 72) male with liver cirrhosis were included. Functional hyposplenism was diagnosed in 28/117 patients (23.9%). Pitted erythrocytes correlated with albumin (p = 0.024), bilirubin (p<0.001), international normalized ratio (INR; p = 0.004), model of end-stage liver disease (MELD) score (p<0.001) and liver stiffness (p = 0.011). Patients with functional hyposplenism had higher MELD scores (median 13 vs. 10; p = 0.021), liver stiffness (46.4 kPa vs. 26.3 kPa; p = 0.011), INR (1.3 vs. 1.2; p = 0.008) and a higher Child-Pugh stage (Child C in 32.1% vs. 11.2%; p = 0.019) as compared to patients without functional hyposplenism. Functional hyposplenism was not associated with the etiology of cirrhosis. Importantly, 9/19 patients with Child C cirrhosis had functional hyposplenism. CONCLUSION: A quarter of patients with liver cirrhosis and almost 50% of patients with Child C cirrhosis have functional hyposplenism. Functional hyposplenism is associated with poor liver function and the degree of portal hypertension, which is characterized by higher liver stiffness measurements in transient elastography.

Assessment of positive iron balance in end-stage renal disease: Could hepcidin-25 be useful?

INTRODUCTION: The aim of our study was to examine the relationship of hepcidin-25 with red blood cell and reticulocyte indices and to evaluate the diagnostic properties of hepcidin-25 in the assessment of positive iron balance in end-stage renal disease (ESRD) patients. METHODS: Eighty anemic ESRD patients (hemoglobin < 110 g/L) were classified as having iron deficiency (ID, N = 20), iron sufficiency (IS, N = 29), and positive iron balance (PB, N = 31) using the conventional biomarkers for iron status evaluation. Hepcidin-25 was determined by a chemiluminescent direct ELISA. RESULTS: Hepcidin-25 was significantly negatively correlated with the proportion of hypochromic erythrocytes (%HYPO) (P = .034) and immature reticulocyte fraction (P = .010) in ID and with the absolute reticulocyte concentration in ID (P = .048) and PB (P = .040). In multivariate models, hepcidin-25 was independently negatively associated with the mean reticulocyte hemoglobin content (CHr; ß = -0.493, P = .004) and red blood cell size factor (RSf) (ß = -0.334, P = .036) only in the PB group. The best hepcidin-25 value to exclude PB was 66.13 µg/L, showing a sensitivity of 61.3%, a specificity of 75.5%, and an AUC of 0.808. CONCLUSION: Our results suggest that hepcidin-25 levels are independently negatively associated with the iron demand for the most recent erythropoiesis only in PB. Hepcidin-25 performed acceptable in discriminating anemic ESRD patients with positive iron balance and may prove to be a useful additional tool in the evaluation of iron status.

Assessment of Fibrinogen Macromolecules Interaction with Red Blood Cells Membrane by Means of Laser Aggregometry, Flow Cytometry, and Optical Tweezers Combined with Microfluidics.

An elevated concentration of fibrinogen in blood is a significant risk factor during many pathological diseases, as it leads to an increase in red blood cells (RBC) aggregation, resulting in hemorheological disorders. Despite the biomedical importance, the mechanisms of fibrinogen-induced RBC aggregation are still debatable. One of the discussed models is the non-specific adsorption of fibrinogen macromolecules onto the RBC membrane, leading to the cells bridging in aggregates. However, recent works point to the specific character of the interaction between fibrinogen and the RBC membrane. Fibrinogen is the major physiological ligand of glycoproteins receptors IIbIIIa (GPIIbIIIa or αIIßß3 or CD41/CD61). Inhibitors of GPIIbIIIa are widely used in clinics for the treatment of various cardiovascular diseases as antiplatelets agents preventing the platelets' aggregation. However, the effects of GPIIbIIIa inhibition on RBC aggregation are not sufficiently well studied. The objective of the present work was the complex multimodal in vitro study of the interaction between fibrinogen and the RBC membrane, revealing the role of GPIIbIIIa in the specificity of binding of fibrinogen by the RBC membrane and its involvement in the cells' aggregation process. We demonstrate that GPIIbIIIa inhibition leads to a significant decrease in the adsorption of fibrinogen macromolecules onto the membrane, resulting in the reduction of RBC aggregation. We show that the mechanisms underlying these effects are governed by a decrease in the bridging components of RBC aggregation forces.

Comparative Assessment of Genotoxic Impacts Induced by Zinc Bulk- and Nano-Particles in Nile tilapia, Oreochromis niloticus.

Fish were separately exposed to 1/2 LC50/96 h values of bulk-Zn and nano-Zn for 7, 14, and 28 days. The induction of micronuclei (MN) and other eight nuclear abnormalities in erythrocytes showed marked time and size dependence. The frequencies of all nuclear anomalies were progressively elevated (p < 0.05) with increasing the time of exposure to both bulk-Zn and nano-Zn. Throughout the study periods, fish exposed to nano-Zn showed the maximum elevation in all studied nuclear anomalies. Based on the fragmented DNA values, both Zn forms induced tissue-specific DNA damage as following gills > liver > muscles. Moreover, nano-Zn exposed groups revealed a maximum percentage of DNA damage among all studied groups, especially after 14 days. The percentage of DNA damage was decreased in all tissues on the 28th day, which reflected the presence of an effective repair mechanism. Finally, nano-Zn exhibited more genotoxic effects than that of its bulk counterparts.

Applications of imaging flow cytometry in the diagnostic assessment of red cell membrane disorders.

BACKGROUND: Red cell membranopathies refers to phenotypically and morphologically heterogeneous disorders. High throughput imaging flow cytometry (IFC) combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy to produce high content image analysis with quantitative analysis. We have evaluated the applications of IFC to examine both the morphology as well as fluorescence signal intensity in red cell membranopathies. METHODS: Fluorescence intensity of eosin-5-maleimide (EMA) labeled red cells was measured for diagnosis of RBC membrane protein defect on Amnis ImageStreamX followed by Image analysis on IDEAS software to study features such as circularity and shape ratio. RESULTS: The hereditary spherocytosis (HS) group showed significantly decreased MFI (52,800 ± 9,100) than normal controls (81,100 ± 4,700) (p < .05) whereas non-HS showed 78,300 ± 9,900. The shape ratio of hereditary elliptocytosis (HE) was significantly higher (43.8%) than normal controls (14.6%). The circularity score is higher in HS (64.15%) than the normal controls (44.3%) whereas the circularity score was very less in HE (10%) due to the presence of elliptocytes. CONCLUSIONS: The advantages of the IFC over standard flow cytometry is its ability to provide high-content image analysis and measurement of parameters such as circularity and shape ratio allow discriminating red cell membranopathies (HS and HE) due to variations in shape and size. It could be a single, effective, and rapid IFC test for detection and differentiation of red cell membrane disorders in hematology laboratories where an IFC is available.

Assessment of hematological, hepato-renal, antioxidant, and hormonal responses of Clarias gariepinus exposed to sub-lethal concentrations of oxyfluorfen.

Little is known about the effects of oxyfluorfen, a diphenyl ether herbicide, exposure on the African catfish (Clarias gariepinus) health. Consequently, the existing investigation was designed to highlight the impacts of oxyfluorfen exposure on C. gariepinus hematological indices, liver and kidney functions, reproductive hormones, and oxidative status. Furthermore, a consequent 10-day depuration period was adopted to evaluate the recovery of the disturbed indicators to normal values. In the first experiment, the 96-h lethal concentration 50 (LC50) of oxyfluorfen for C. gariepinus was determined using probit analysis. Next, in a second experiment, 180 healthy fish (average initial body weight: 164.23 ±â€¯0.24) were randomly assigned to 4 experimental groups exposed to 0, 1/10, 1/8, or 1/5 96-h LC50 of oxyfluorfen. The hematological profile, hepatic enzymes, kidney damage byproducts, reproductive hormones, oxidative stress, and lipid peroxidation indicators together with acetylcholinesterase (AChE) content were assessed. A histopathological examination of the hepatic, renal, brain, and testicular tissues was accomplished. Moreover, the expression of the oxidative stress-related gene was carried out. The results showed that 96-h LC50 of oxyfluorfen for C. gariepinus was 11.698 mg/L. Exposure to sublethal levels of oxyfluorfen induced macrocytic hypochromic anemia, leukopenia, lymphopenia, monocytopenia, and eosinopenia. Also, a concentration-dependent increase in alanine transaminase, alkaline phosphatase, aspartate transaminase, urea, creatinine, catalase, and malondialdehyde was detected following oxyfluorfen exposure together with upregulation of catalase gene. But, significant concentration-dependent reductions in AChE, glutathione transferase, reduced to oxidized glutathione ratio, estradiol, and testosterone activities were recorded. These biochemical alterations were accompanied by pathological perturbations in hepatic, renal, brain, and testicular tissues. Following 10 days of recovery, only the hematological impairments were abolished. Conclusively, the herbicides oxyfluorfen could induce multiple negative impacts on C. gariepinus with oxidative stress as a probable underlying mechanism. Additionally, a recovery period of 10 days was not enough to restore these impairments.

Machine learning in multiexposure laser speckle contrast imaging can replace conventional laser Doppler flowmetry.

Laser speckle contrast imaging (LSCI) enables video rate imaging of blood flow. However, its relation to tissue blood perfusion is nonlinear and depends strongly on exposure time. By contrast, the perfusion estimate from the slower laser Doppler flowmetry (LDF) technique has a relationship to blood perfusion that is better understood. Multiexposure LSCI (MELSCI) enables a perfusion estimate closer to the actual perfusion than that using a single exposure time. We present and evaluate a method that utilizes contrasts from seven exposure times between 1 and 64 ms to calculate a perfusion estimate that resembles the perfusion estimate from LDF. The method is based on artificial neural networks (ANN) for fast and accurate processing of MELSCI contrasts to perfusion. The networks are trained using modeling of Doppler histograms and speckle contrasts from tissue models. The importance of accounting for noise is demonstrated. Results show that by using ANN, MELSCI data can be processed to LDF perfusion with high accuracy, with a correlation coefficient R = 1.000 for noise-free data, R = 0.993 when a moderate degree of noise is present, and R = 0.995 for in vivo data from an occlusion-release experiment.

Human erythrocyte band 3 is a host receptor for Plasmodium falciparum glutamic acid-rich protein.

Malaria remains a major global threat to human health and economic development. Microvascular lesions caused by Plasmodium falciparum-infected human erythrocytes/red blood cells are hallmarks of severe pathogenesis contributing to high mortality, particularly in children from sub-Saharan Africa. In this study, we used a phage display complementary DNA library screening strategy to identify P falciparum glutamic acid-rich protein (PfGARP) as a secreted ligand that recognizes an ectodomain of human erythrocyte anion-exchanger, band 3/AE1, as a host receptor. Domain mapping of PfGARP revealed distinct nonoverlapping repeats encoding the immune response epitopes and core erythrocyte-binding activity. Synthetic peptides derived from the erythrocyte-binding repeats of PfGARP induced erythrocyte aggregation reminiscent of the rosetting phenomenon. Using peptides derived from the immunogenic repeats, a quantitative immunoassay was developed to detect a selective immune response against PfGARP in human plasma samples obtained from patients in rural Mali, suggesting the feasibility of PfGARP as a potential biomarker of disease progression. Collectively, our results suggest that PfGARP may play a functional role in enhancing the adhesive properties of human erythrocytes by engaging band 3 as a host receptor. We propose that immunological and pharmacological inhibition of PfGARP may unveil new therapeutic options for mitigating lesions in cerebral and pregnancy-associated malaria.

In vitro assessment of the toxicity of small silver nanoparticles and silver ions to the red blood cells.

This work reports the toxicity of small silver nanoparticles (nanoAg, 20 nm) and silver ions (Ag+) to the red blood cells with the silver concentration level of 10-6 g/mL. Results show that red blood cells (RBCs) start hemolysis when treated by nanoAg of 1.5 × 10-5 g/mL or Ag+ of 2.9 × 10-7 g/mL. A low ATPase activity of 30% has been observed after RBCs being treated with Ag+ of 2.6 × 10-7 g/mL, while the nanoAg does not obviously affect the ATPase activity. In molecular level, Ag+ is more toxic to the amino acid residues than nanoAg according to the change of fluorescence characteristics of hemoglobin (Hb). However, the nanoAg has been found to be more toxic than Ag+ to the secondary structure of Hb in terms of the loss of α-helix content.

Genotoxic and mutagenic assessment of iron oxide (maghemite-γ-Fe2O3) nanoparticle in the guppy Poecilia reticulata.

The environmental risk of nanomaterials (NMs) designed and used in nanoremediation process is of emerging concern, but their ecotoxic effects to aquatic organism remains unclear. In this study, the citrate-coated (maghemite) nanoparticles (IONPs) were synthesized and its genotoxic and mutagenic effects were investigated in the female guppy Poecilia reticulata. Fish were exposed to IONPs at environmentally relevant iron concentration (0.3 mg L-1) during 21 days and the animals were collected at the beginning of the experiment and after 3, 7, 14 and 21 days of exposure. The genotoxicity and mutagenicity were evaluated in terms of DNA damage (comet assay), micronucleus (MN) test and erythrocyte nuclear abnormalities (ENA) frequency. Results showed differential genotoxic and mutagenic effects of IONPs in the P. reticulata according to exposure time. The IONP induced DNA damage in P. reticulata after acute (3 and 7 days) and long-term exposure (14 and 21 days), while the mutagenic effects were observed only after long-term exposure. The DNA damage and the total ENA frequency increase linearly over the exposure time, indicating a higher induction rate of clastogenic and aneugenic effects in P. reticulata erythrocytes after long-term exposure to IONPs. Results indicated that the P. reticulata erythrocytes are target of ecotoxicity of IONPs.

[Assessment of Erythropoiesis Status in Roach (Rutilus rutilus) of the Radioactively Contaminated Techa River].

At present volumetric activity of ß-emitting radionuclides in water at various locations of the Techa River ranges from 5 to 40-Bq/L; a specific activity of ß-emitting radionuclides in the bottom sediments at various locations ranges 10 Ito 106 Bq/kg dry weight. A significant increase of the erythroblast content in blood as compared to that in the roach from the reference watercourse (the Miass River) was observed during spawning in the spring. Due to this fact the number of erythrocytes was equal to that in the control animals under chronic radiation exposure at the dose rates of 0.9 and 16 µGy/day, and was insufficient at the dose rate of 108 gGy/day. During summer feeding no changes in the indexes of erythropoiesis in roach were observed under chronic radiation exposure at the dose rate of 0.9 µGy/day; the number of erythrocytes in the peripheral blood declines when the dose rates are 16 and 108 µGy/day. When performing a regression analysis, we revealed a dose-rate-dependent decrease in the absolute number of erythrocytes, normocytes, polychromatocytes, dividing and non-dividing erythroid cells in the peripheral blood of roach from the Techa River and an increase of a relative number of normochromatophylic erythrocytes.

MIF-Mediated Hemodilution Promotes Pathogenic Anemia in Experimental African Trypanosomosis.

Animal African trypanosomosis is a major threat to the economic development and human health in sub-Saharan Africa. Trypanosoma congolense infections represent the major constraint in livestock production, with anemia as the major pathogenic lethal feature. The mechanisms underlying anemia development are ill defined, which hampers the development of an effective therapy. Here, the contribution of the erythropoietic and erythrophagocytic potential as well as of hemodilution to the development of T. congolense-induced anemia were addressed in a mouse model of low virulence relevant for bovine trypanosomosis. We show that in infected mice, splenic extramedullary erythropoiesis could compensate for the chronic low-grade type I inflammation-induced phagocytosis of senescent red blood cells (RBCs) in spleen and liver myeloid cells, as well as for the impaired maturation of RBCs occurring in the bone marrow and spleen. Rather, anemia resulted from hemodilution. Our data also suggest that the heme catabolism subsequent to sustained erythrophagocytosis resulted in iron accumulation in tissue and hyperbilirubinemia. Moreover, hypoalbuminemia, potentially resulting from hemodilution and liver injury in infected mice, impaired the elimination of toxic circulating molecules like bilirubin. Hemodilutional thrombocytopenia also coincided with impaired coagulation. Combined, these effects could elicit multiple organ failure and uncontrolled bleeding thus reduce the survival of infected mice. MIF (macrophage migrating inhibitory factor), a potential pathogenic molecule in African trypanosomosis, was found herein to promote erythrophagocytosis, to block extramedullary erythropoiesis and RBC maturation, and to trigger hemodilution. Hence, these data prompt considering MIF as a potential target for treatment of natural bovine trypanosomosis.

Detection and quantification of subtle changes in red blood cell density using a cell phone.

Magnetic levitation has emerged as a technique that offers the ability to differentiate between cells with different densities. We have developed a magnetic levitation system for this purpose that distinguishes not only different cell types but also density differences in cells of the same type. This small-scale system suspends cells in a paramagnetic medium in a capillary placed between two rare earth magnets, and cells levitate to an equilibrium position determined solely by their density. Uniform reference beads of known density are used in conjunction with the cells as a means to quantify their levitation positions. In one implementation images of the levitating cells are acquired with a microscope, but here we also introduce a cell phone-based device that integrates the magnets, capillary, and a lens into a compact and portable unit that acquires images with the phone's camera. To demonstrate the effectiveness of magnetic levitation in cell density analysis we carried out levitation experiments using red blood cells with artificially altered densities, and also levitated those from donors. We observed that we can distinguish red blood cells of an anemic donor from those that are healthy. Since a plethora of disease states are characterized by changes in cell density magnetic cell levitation promises to be an effective tool in identifying and analyzing pathologic states. Furthermore, the low cost, portability, and ease of use of the cell phone-based system may potentially lead to its deployment in low-resource environments.

Assessment of the relationship between red blood cell distribution width and preganecy hypertension disease.

AIMS: The aim of this study was to explore the correlation between red blood cell distribution width (RDW) and Preganecy Heypertion Disease (PHD) . METHODS: A retrospective study was carried out involving 149 pregnancies with PHD(67 gestational hypertension, 39 Mild preeclampsia, 24 severe preeclampsia and 19 eclampsia) and 70 health pregnant women as controls. The data of RDW were reviewed from antenatal and delivery records. Explored the correlation between RDW and PDH through Logistic Regression analysis, analyzed the clinical value of RDW to predict PHD by receiver operating characteristic (ROC) curve analysis. RESULTS: RDW in different gestational time(20th week, 24th week, 28th week) of different pregnant women groups had differences (P < 0.05), but pregnant women in the same group had no difference from 20th week to 28th week(P > 0.05). Logistic regression analysis showed that RDW was a risk factor for PHD (odds ratio 2.683; 95 % confidence interval 1.472-6.096), the optimal RDW-CV threshold was 14.1 % to predict PHD by the ROC curve, the sensitivity and speciality were 72.5 % and 77.9 %. CONCLUSIONS: RDW as a new chronic inflammation mediator, which was a high-risk factor of PHD, also had certain clinical value to predict the occurrence of PHD.

Intravital Microscopy Imaging of the Liver following Leishmania Infection: An Assessment of Hepatic Hemodynamics.

Intravital microscopy (IVM) is a powerful optical imaging technique that has made possible the visualization, monitoring and quantification of various biological events in real time and in live animals. This technology has greatly advanced our understanding of physiological processes and pathogen-mediated phenomena in specific organs. In this study, IVM is applied to the mouse liver and protocols are designed to image in vivo the circulatory system of the liver and measure red blood cell (RBC) velocity in individual hepatic vessels. To visualize the different vessel subtypes that characterize the hepatic organ and perform blood flow speed measurements, C57Bl/6 mice are intravenously injected with a fluorescent plasma reagent that labels the liver-associated vasculature. IVM enables in vivo, real time, measurement of RBC velocity in a specific vessel of interest. Establishing this methodology will make it possible to investigate liver hemodynamics under physiological and pathological conditions. Ultimately, this imaging-based methodology will be important for studying the influence of L. donovani infection on hepatic hemodynamics. This method can be applied to other infectious models and mouse organs and might be further extended to pre-clinical testing of a drug's effect on inflammation by quantifying its effect on blood flow.

In Vitro Comparative Assessment of Mechanical Blood Damage Induced by Different Hemodialysis Treatments.

Gradual deterioration of red blood cells (RBCs) due to mechanical stress (chronic hemolysis) is unavoidable during treatments that involve extracorporeal blood circulation, such as hemodialysis (HD). This effect is generally undetectable and does not generate any acute symptoms, but it leads to an increase in plasma free hemoglobin (fHb). There are no absolute safety levels for fHb increase, indicating the need for an empirical evaluation using comparative testing. The increase in fHb levels was investigated in vitro by applying double-needle double-pump HD (HD-DNDP), a new modality in which arterial and venous pumps both run continuously. fHb was measured during typical and worst-case simulated dialysis treatments (double-needle single-pump HD [HD-DNSP], hemodiafiltration [HDF-DN], single-needle double-pump HD [HD-SNDP], and HD-DNDP) performed in vitro using bovine blood for 4 h. Hemolysis-related indices (fHb%; index of hemolysis, IH; and normalized IH) were calculated and used for comparison. The increase in fHb during either HDF-DN or HD-SNDP with Artis and AK200 dialysis machines was similar, while the fHb at the maximum real blood flow rate (Qbreal ) at the completion of the HD-DNDP treatment on Artis was higher than that for HD-DNSP using a Phoenix dialysis machine (fHb% = 1.24 ± 0.13 and 0.92 ± 0.12 for the Artis machine with HD-DNDP at Qbreal = 450 mL/min and Phoenix with HD-DNSP at Qbreal = 500 mL/min, respectively). However, the fHb levels increased linearly, and no steep changes were observed. The increases observed during HD-DNDP were the same order of magnitude as those for widely used bloodlines and treatment modes for delivering dialysis treatments. The observed results matched literature findings, and thus the measured fHb trends are not predicted to have clinical side effects. HD-DNDP treatment with Artis does not merit any additional concern regarding mechanical stress to RBCs compared with that observed for routinely used dialysis treatments, bloodlines and machines. Although the in vitro measurement of the fHb increase in bovine blood does not allow a prediction of the absolute level of blood mechanical damage or the possible effects in humans, such measurements are valuable for assessing hemolytic harm by performing tests comparing the proposed treatment with existing devices.

In vivo genotoxicity and cytotoxicity assessment of cadmium chloride in peripheral erythrocytes of Labeo rohita (Hamilton).

Cadmium chloride (CdCl2) induced genotoxicity and cytotoxicity has been assessed in the peripheral blood erythrocytes of freshwater fish Labeo rohita exposed to 0.37 and 0.62mg/L of CdCl2 in water for 100 days. The blood samples of the fish were collected at different intervals (days 1, 3, 5, 10, 15, 30, 60 and 100) of exposure period to analyze DNA damage using comet assay and the occurrence of micronuclei and other cellular anomalies. The results of comet assay showed a significant increase in the mean percentage of tail DNA at both the concentrations. Exposure to CdCl2 also induced micronuclei in addition to many nuclear abnormalities such as nuclear bud, binucleates, lobed, notched and vacuolated nuclei. Cytoplasmic abnormalities like echinocytes, acanthocytes, notched, microcytes and cells with vacuolated cytoplasm were also observed. The metal exposed groups showed significant variation in the frequency of cellular abnormalities as well as the extent of DNA damage in comparison to controls. These frequencies increased significantly (p<0.05) in concentration dependent manner, peaking on 10th day while a decreasing trend was observed after 15 days of the exposure period.

An analysis of the NIH-supported sickle cell disease research portfolio.

Sickle cell disease (SCD), an inherited blood disorder is due to a single amino acid substitution on the beta chain of hemoglobin, and is characterized by anemia, severe infections, acute and chronic pain, and multi-organ damage. The National Institutes of Health (NIH) is dedicated to support basic, translational and clinical science research to improve care and ultimately, to find a cure for SCD that causes such suffering. This report provides a detailed analysis of grants funded by the NIH for SCD research in Fiscal Years 2007 through 2013. During this period, the NIH supported 247 de novo grants totaling $272,210,367 that address various aspects of SCD. 83% of these funds supported research project grants investigating the following 5 scientific themes: Pathology of Sickle Red Blood Cells; Globin Gene Expression; Adhesion and Vascular Dysfunction; Neurological Complications and Organ-specific Dysfunction; and Pain Management and Intervention. The remaining 17% of total funds supported career development and training grants; Small Business Innovation Research (SBIR) and Small Business Technology Transfer (STTR) grants; large Center grants; and Conference grants. Further analysis showed that the National Heart, Lung, and Blood Institute (NHLBI) is the largest funder of SCD research within NIH with 67% of total grants, contributing 77% of total funds; followed by the National Institute for Digestive Diseases and Kidney (NIDDK) that is funding 19% of grants, contributing 13% of total funds. The remaining 14% of grants totaling 10% of the funds were supported by all other NIH Institutes/Centers (ICs) combined. In summary, the NIH is using multiple funding mechanisms to support a sickle cell disease research agenda that is intended to advance the detection, treatment, and cure of this debilitating genetic disease.