Standard Complete Blood Count Research Articles

Value of Red Cell Distribution Width in Children with Congenital Heart Disease Associated- Pulmonary Arterial Hypertension

Background: Pulmonary arterial hypertension (PAH) commonly occurs as a consequence following untreated congenital heart disease (CHD). It’s often related to high morbidity and mortality rates. The work was aimed at assessing the RDW measurements in children who had PAH-CHD.
 Methods: Thirty children who have PAH-CHD took part in this study. Electrocardiography and echocardiographic evaluation were applied to all participants. RDW, a parameter included in complete blood count, was measured.
 Results: Our study involved thirty children. Up to 53.3% of them were males, whose age varied between three months and twelve months with median of 5 months. The optimal cutoff RDW measurement reached 17.0% (mean); a significant relation between RDW level and low oxygen saturation was reported (p=0.02), right ventricular fractional area change (p=0.023), and left ventricular ejection fraction (p=0.13).
 Conclusion: RDW, a parameter included in the standard complete blood count, showed a sagnificant elevation in children who had PAH-CHD children. In addition, it was significantly correlated with hypoxia and right ventricular fractional area change and left ventricular ejection fraction.

The effect of nasal septum deviation type on the systemic inflammatory index and blood markers of inflammation

Aims: The aim of this study was to identify the changes in blood markers of inflammation and the systemic immune inflammatory index in patients with nasal septum deviation (NSD). An additional aim was to determine if there was a significant difference in the findings according to septum deviation type.
 Methods: This retrospective study included 321 patients diagnosed with NSD via physical examination and CT (NSD group), and 101 healthy controls. Standard complete blood count was performed. Main blood parameters and mean platelet volume (MPV), the neutrophil-to-lymphocyte ratio (NLR), the platelet-to-lymphocyte ratio (PLR), and the systemic immune inflammatory index (SII) were recorded. NSD was classified as 3 types according to Dreher classification, and the types were compared to each other and the control group.
 Results: The hemoglobin level, and platelet and neutrophil counts were significantly higher in the NSD group than in the control group (P

A-144 Three-wavelength Polarization Imaging Method for the Analysis of Individual Human Blood Cells With Dye-Enhanced Dispersion of Dilution Buffer

Abstract Background Performing basic hematology analysis in a near-patient setting enables more rapid clinical decision making in the process of diagnosis. Traditional blood analyzers in central labs use a complex combination of sensing principles and chemical modification of the sample to derive the necessary parameters for a complete blood count (CBC). To reduce cost and maintenance, varying approaches have been employed to determine a CBC with a single-step measurement technique. One common option is the use of optical microscopy to deliver highly reliable results, examples are fluorescence microscopy, digital holographic microscopy, or spatial light interference microscopy. However, most of these technologies lack a proper determination of cellular-level RBC parameters. Therefore, a non-interferometric, quantitative-phase measurement technique was developed to derive relevant CBC parameters from a single measurement, comparable to a standard lab analyzer. Methods The optical setup employed by this method enables the parallel acquisition of brightfield and differential interference contrast (DIC) images by polarization-sensitive cameras. By using three cameras with individual optical filters, quantitative phase and absorption information can be calculated, which enables the determination of mean cellular volume (MCV), mean cellular hemoglobin (MCH), and mean cellular hemoglobin concentration (MCHC) for red blood cells (RBCs). The whole-blood sample is diluted in a buffer containing a non-cell-penetrating dye to enhance optical refraction contrast and therefore precision in the dependent calculated quantities. The sample is then introduced to a glass flow cell, and the cells are allowed to sediment to the bottom for imaging. Machine learning is applied to the derived images to classify unstained native white blood cells (WBCs) to yield a 5-part differential, which distinguishes between neutrophil, monocyte, lymphocyte, eosinophil and basophil. The algorithm is trained by imaging FACS-validated, purified WBC subgroups. All blood samples are measured on an ADVIA® 2120 Hematology System for control. Results A one-step, optical CBC hematology system based on quantitative phase and absorption image analysis was developed. The correlations for MCV and MCHC compared to a standard lab CBC analyzer were 0.77 and 0.84. The accuracy for the 5-part differential WBC classification was approximately 0.8. To enable a visual inspection of the classification results, a pseudo-colored image of each cell was generated and uploaded in a review tool. Conclusion With a more-refined, less-expensive optical setup, a high-quality CBC analyzer suitable for use at the point of care can be derived with the presented method. A possible simplification could be the use of low-cost but high-resolution Fourier ptychography microscopy (FPM) technology. Imaging of blood samples on a single-cell level offers the potential for the analysis of even more morphological parameters, for example, disease-related changes in the cell membrane. Also, the adhesion behavior of the WBCs in the glass flow cell could indicate cell activation status and therefore give information about infection processes. *Trademark disclaimer: “ADVIA, and all associated marks are trademarks of Siemens Healthcare Diagnostics, Inc. or its affiliates. All other trademarks and brands are the property of their respective owners.”

Relationship of red cell distribution width-coefficient of variance with the severity of systolic dysfunction in patients of heart failure with reduced ejection fraction

Background: Red blood cells distribution width (RDW)-coefficient of variance (RDW-CV) is a measure of red blood cell width variation, reported as part of a standard complete blood count. Although it is usually measured as a routine test, actually its values have been only used in the differential diagnosis of anemia and its high values indicate the presence of anisocytosis. It has been demonstrated that RDW could be considered as an independent additional marker of cardiovascular events in patients with heart failure with reduced ejection fraction. However, the role of RDW in patients with heart failure is less known. This study was designed to evaluate the relationship between RDW-CV and severity of systolic dysfunction in patients with heart failure with reduced ejection fraction (HFrEF). Methods: This cross-sectional study was carried out at the Department of Cardiology, Sir Salimullah Medical College and Mitford Hospital, Dhaka, Bangladesh from April 2019 to March 2020. This study included 100 consecutive patients with dyspnea, fatigue and fluid retention suggestive of heart failure. Participating subjects were subjected to a detailed history taking, complete physical examination, New York Heart Association (NYHA) class, 12-lead electrocardiogram (ECG) and routine laboratory investigations including RDW-CV calculate from complete blood count were performed. Left ventricular ejection fraction (LVEF) was measured by echocardiography. Results: The study showed statistically significant negative correlation between RDW-CV and LVEF (r = -0.498, p value<0.001). Conclusion: In the study showed that elevated RDW was associated with decreased ejection fraction assessed by echocardiography in patients of HFrEF. Therefore, increased RDW could be used as an additional marker for identifying severity of left ventricular dysfunction. BIRDEM Med J 2023; 13(3): 124-129

Estimation of cell type proportions from bulk RNA-Seq of porcine whole blood samples using partial reference-free deconvolution

Whole blood has become increasingly utilized in transcriptomic studies because it is easily accessible and can be collected from live animals with minimal invasiveness. However, whole blood represents an extremely complex mixture of cell types, and cell type proportions can confound downstream statistical analyses. Information on cell type proportions may be missing from blood transcriptome studies for a variety of reasons. Experimental approaches for cell counting, such as cell sorting, are arduous and expensive, and therefore may not feasible for studies conducted on a limited budget. Statistical deconvolution can be applied directly to transcriptomic data sets to estimate cell type proportions. In addition to being financially advantageous, computational deconvolution can readily be applied to old datasets, where it may be difficult or impossible to re-analyze for cell type information. In an effort to assist researchers in recovering cell type proportions from porcine whole blood transcriptome samples, we present a manually curated set of porcine blood cell markers that can be utilized in a partial reference-free deconvolution framework to obtain estimates of cell types measured in a standard complete blood count (CBC) panel, which includes neutrophils, lymphocytes, monocytes, eosinophils, basophils, and red blood cells.

Relationship between hematologic parameters related to systemic inflammation and insulin resistance-associated metabolic parameters in women with polycystic ovary syndrome.

The aim of the present study was to evaluate the associations between hematologic parameters related to systemic inflammation and insulin resistance-associated metabolic parameters in women with polycystic ovary syndrome (PCOS). Eighty-two women between the ages of 18 and 35 years who were diagnosed with PCOS were included in this study. A 2-hour 75-g oral glucose tolerance test (OGTT) was administered to all study participants; fasting and postprandial glucose and insulin levels were measured simultaneously during the 2-hour OGTT. Hematologic parameters were derived from a standard complete blood count and a differential count of fasting-state blood samples. The correlations between hematologic parameters and insulin resistance-associated clinical and metabolic parameters were evaluated using the Spearman rank correlation and partial correlation coefficients. Hematologic parameters related to systemic inflammation were compared between the two groups, categorized by the presence or absence of insulin resistance. Significant differences in the absolute neutrophil count, absolute monocyte count, platelet count, and neutrophil-lymphocyte ratio were found between the insulin-resistant group and insulin-nonresistant group. Correlation analysis found that all hematological parameters, except for the platelet-lymphocyte ratio, were associated with at least one insulin resistance-associated metabolic parameter. However, these significant correlations between hematological and metabolic parameters were attenuated after controlling for the effects of other covariates using partial correlation analysis. The association between hematologic parameters indicative of systemic inflammation and insulin resistance-associated metabolic parameters seems to be strongly influenced by other anthropometric covariates in women with PCOS.

Abstract 2292: Novel method for the detection and evaluation of disease biomarkers on white blood cells from liquid biopsies

Abstract Purpose: Blood based liquid biopsies of circulating tumor cells (CTCs) are a non-invasive tool with proven effectiveness in the prognosis, treatment selection, and stratification of cancer patients. In this study we demonstrate a novel application of the RareCyte platform, which was developed to find and analyze rare CTCs from blood samples, in evaluating white blood cell (WBC) sub-populations for disease biomarkers. Experimental Procedures: Using AccuCyte®, an unbiased density-based method for collecting nucleated cells from whole blood, we transferred the nucleated cells to slides and stained for CD14, CD66b, and CK/EpCAM to identify monocytes (CD14+/CD66b-), granulocytes (CD14-/CD66b+), and CTCs (CK/EpCAM+), as well as a drug-target biomarker to evaluate its expression across all cells. These slides were then imaged with CyteFinder®, an automated multiparameter immunofluorescent (IF) microscopy system that applies machine learning algorithms for cell identification. Each slide contains millions of cells, so we developed novel scanning and analysis algorithms to quantify WBC sub-populations in addition to rare CTCs. Results: In addition to detection and enumeration of CTCs, the RareCyte system was able to detect individual white blood cells and quantify monocyte and granulocyte subpopulations. Our results correlated very well with standard CBC (Complete Blood Count) readings from multiple donors. Additionally, the monocyte and granulocyte populations were evaluated for expression of a drug-target biomarker. While expression was observed on both granulocytes and monocytes, monocytes had a significantly higher expression of the drug-target biomarker, a critical finding for future implementation of this drug. Conclusions: Our results show that through modifications to the RareCyte platform we can identify rare and non-rare cell populations simultaneously and evaluate expression of biomarkers and drug-targets across multiple cells of interest. This new application combines the benefits of high throughput data analysis of flow cytometry with the specificity and sensitivity of rare cell detection. Additionally, high-resolution multiplexed immunofluorescent imaging provides cell morphology and biomarker localization information. Our ability to add multiple biomarkers in conjunction with CTC and other rare cell detection provides multiple avenues for future research. One possible application is characterizing white blood cells that cluster with CTCs, which are known to portend a worse prognosis for patients. Our modified workflow could be implemented to precisely characterize these WBCs that contribute to the metastatic process. Utilization of the RareCyte platform for both rare and non-rare cell analysis presents an exciting opportunity for a deeper evaluation of patients and their prognosis through liquid biopsies. Citation Format: Erin Bayer, Jon Ladd, Joshua Nordberg, Arturo B. Ramirez, Jessica A. Baker, Thomas Daly. Novel method for the detection and evaluation of disease biomarkers on white blood cells from liquid biopsies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2292.

Hemoglobin Determination Using Pulse Co-Oximetry and Reduced-Volume Blood Gas Analysis in the Critically Ill: A Prospective Cohort Study.

Hospital-acquired anemia is common in patients hospitalized in the intensive care unit (ICU). A major source of iatrogenic blood loss in the ICU is the withdrawal of blood for laboratory testing. The aim of our study was to analyze the feasibility and accuracy of non-invasive spot-check pulse co-oximetry (SpHb), and a reduced-volume blood gas analysis (ABG Hb) for the determination of Hb concentration in critically ill patients. Comparisons between Hb determined with test devices and the gold standard-complete blood count (CBC)-were performed using Bland-Altman analysis and concordance correlation coefficient (CCC). The limits of agreement between SpHb and CBC Hb were -2.0 [95%CI -2.3-(-1.7)] to 3.6 (95%CI 3.3-3.9) g/dL. The limits of agreement between ABG Hb and CBC Hb were -0.6 [95%CI -0.7-(-0.4)] to 2.0 (95%CI 1.9-2.2) g/dL. Spearman's coefficient and CCC between ABG Hb and CBC Hb were 0.96 (95%CI 0.95-0.97, p < 0.001) and 0.91 (95%CI 0.88-0.92), respectively. Non-invasive spot-check Hb co-oximetry is not sufficiently accurate for the monitoring of hemoglobin concentration in critically ill patients. Reduced volume arterial blood gas analysis has acceptable accuracy and could replace complete blood count for the monitoring of Hb concentration in critically ill patients, leading to a significant reduction in blood volume lost for anemia diagnostics.

Comparison of two point-of-care measurements of hemoglobin against standard laboratory values during pediatric liver transplantation: A retrospective review.

Point-of-care hemoglobin testing devices play an important role in intraoperative anesthetic management where significant hemorrhage is anticipated; however, the reliability of these devices has not been examined in the context of pediatric liver transplantation. In this retrospective observational study, we aimed to determine whether 95% of results from two point-of-care hemoglobinometers, the HemoCue and iSTAT, would fall within a difference of ±1g/dl, our a priori-defined clinically acceptable level of agreement, of the hemoglobin measures on a core laboratory complete blood count. We retrospectively collected data from 70 patients presenting for a liver transplant at a single center, tertiary care pediatric hospital over a 3.5-year period. We analyzed 92 contemporaneous pairs of hemoglobin values from the HemoCue and complete blood count, and 252 pairs of hemoglobin values from the iSTAT and complete blood count. Agreement between the point-of-care devices and complete blood count was assessed using Bland-Altman analysis, which was the primary outcome. Secondary analyses included an error grid analysis and Cohen's kappa statistic. Both point-of-care devices underestimated complete blood count hemoglobin values and neither device satisfied our a priori-defined clinically acceptable level of agreement that 95% of values would fall within ±1g/dl of the complete blood count measurement. The mean difference [limits of agreement] of the HemoCue was 0.4g/dl (p < .001) [-0.9 to 1.6g/dl] and of the iSTAT was 0.6g/dl (p < .001) [-1.4to 2.6g/dl]. Secondary error grid analysis revealed that neither device performed well enough to replace a complete blood count at critical thresholds of hemoglobin values. While the HemoCue and iSTAT contribute information in a timely manner during dynamic intraoperative situations, there is significant imprecision compared to the gold standard complete blood count. If clinical stability allows, the results of these point-of-care hemoglobinometers should be confirmed with a complete blood count, rather than being used as the sole factor in determining transfusion needs during pediatric liver transplantation.

Toxicity Assessment of Mesoporous Silica Nanoparticles upon Intravenous Injection in Mice: Implications for Drug Delivery.

Nanoparticles are popular tools utilized to selectively deliver drugs and contrast agents for identification and treatment of disease. To determine the usefulness and translational potential of mesoporous silica nanoparticles (MSNs), further evaluations of toxicity are required. MSNs are among the most utilized nano-delivery systems due to ease of synthesis, pore structure, and functionalization. This study aims to elucidate toxicity as a result of intravenous injection of 25 nm MSNs coated with chitosan (C) or polyethylene glycol (PEG) in mice. Following acute and chronic injections, blood was evaluated for standard blood chemistry and complete blood count analyses. Blood chemistry results primarily indicated that no abnormalities were present following acute or chronic injections of MSNs, or C/PEG-coated MSNs. After four weekly administered treatments, vital organs showed minor exacerbation of pre-existing lesions in the 35KPEG-MSN and moderate exacerbation of pre-existing lesions in uncoated MSN and 2KPEG-MSN treatment groups. In contrast, C-MSN treatment groups had minimal changes compared to controls. This study suggests 25 nm MSNs coated with chitosan should elicit minimal toxicity when administered as either single or multiple intravenous injections, but MSNs coated with PEG, especially 2KPEG may exacerbate pre-existing vascular conditions. Further studies should evaluate varying sizes and types of nanoparticles to provide a better overall understanding on the relation between nanoparticles and in vivo toxicity.

Investigation of the Efficacy of Home-Based Oral Chloroquine Treatment among Under-Five Children with &amp;lt;i&amp;gt;Plasmodium falciparum&amp;lt;/i&amp;gt; Malaria in Some Parts of Jos, Plateau State, Nigeria

Background: Nigeria is currently a malaria endemic country with an estimated 76% of her population at risk of contracting malaria [1]. According to a study in Nigeria, the first line of action mothers took when their children under 5 years have malaria showed that over 50% of them used non-prescription drugs they have at home or bought from pharmacy stores. And 60% of the most commonly used drugs for malaria treatment were chloroquine [2]. Many recent studies have demonstrated re-emergence of chloroquine-sensitive P. falciparum, suggesting a possible role in future malaria control [3]. Objective: The aim of this study was to investigate the effect of home-based oral chloroquine treatment among children under 5 years with Plasmodium falciparum malaria attending Jos University Teaching Hospital and OLA Hospital in Jos Metropolis. Method: This is a cross-sectional study of 93 malaria and non-malaria children. Malaria diagnosis was carried out using microscopical examination of Leishman’s stained thick and thin blood films, P. falciparum parasitemia was assessed by standard microscopy techniques and complete blood count was done using Beckman Coulter Analyzer. Results: The body temperature on admission was significantly lower (p ˚C ± 0.07˚C) than in the three malaria groups. The mean body temperature of chloroquine treated children with malaria was significantly lower (p Plasmodium falciparum parasitemia and degree of anemia in children under 5 years with Plasmodium falciparum in Jos Metropolis.

Reliability of transcutaneous hemoglobin measurements in an outpatient pediatric clinic

ABSTRACT Objective Our study assessed the reliability of a transcutaneous hemoglobin (tcHgb) measurement as compared to a standard capillary hemoglobin (cHgb) measurement in screening for iron deficiency anemia in a single university-based pediatric outpatient clinic. Methods Study participants included all pediatric patients requiring a hemoglobin (Hgb) assessment from July 2019 to June 2020. A tcHgb measurement was attempted on all children who received a cHgb measurement. Additional variables evaluated were age, visit type, gender, insurance type, weight, BMI percentile and presence of comorbid conditions. Results Of 777 attempts, both cHgb and tcHgb were obtained in 196 children aged 9 months to 21 years. Attempts were most successful in children > 2 years of age due to finger size and ability to remain still for one to two minutes. The mean cHgb was 12.5 ± 1.5 g/dL, mean tcHgb value 13.1 ± 2.1 g/dL, and the mean difference was 0.6 ± 2.1 g/dL (tcHgb-cHgb). An intraclass correlation coefficient was 0.29. There were no differences with regards to age, visit type, gender, insurance type, weight, BMI percentile and presence of comorbid conditions. Bland-Altman analysis displayed a lack of agreement between the tcHgb and cHgb measurements and tcHgb tended to over-estimate Hgb values when the cHgb was low. Conclusions The measurement of a transcutaneous Hgb is less invasive for pediatric patients but has significant limitations. Smaller children (probe specifications) and movement limited the ability to obtain a tcHgb measurement in ~75% of children tested. Falsely normal tcHgb values occurred due to overestimation of hemoglobin when compared to the traditional cHgb device. The suboptimal sensitivity of the tcHgb device may cause providers to miss a diagnosis of anemia. Future research should compare both methods of Hgb assessment to the gold standard laboratory-analyzed complete blood count and use a smaller probe for children under 2, once available.

Complete blood count and cell population data parameters from the Abbott Alinity hq analyzer are useful in differentiating myelodysplastic syndromes from other forms of cytopenia.

Myelodysplastic syndromes (MDS) are characterized by morphologic dysplasia and cytopenia and have a propensity for acute leukemic transformation. However, dysplasia is diagnosed by morphology, thus having cell population data (CPD) that can differentiate cytopenic patients with MDS from other conditions may facilitate accurate diagnosis. We assessed the utility of complete blood count (CBC) parameters and CPD derived from an Abbott Alinity hq analyzer to discriminate MDS-related cytopenia. The patient cohort (n=345) included 64 samples from patients with MDS, 162 from patients with other cytopenia, and 119 from healthy controls. The hematological parameters and research use-only parameters of the Abbott Alinity hq analyzer were compared between the cytopenic groups. The effectiveness of the individual standard and research CBC parameters to differentiate MDS from other forms of cytopenia was assessed through a receiver operating characteristics (ROC) analysis. The percentage of MAC (Macrocytic RBCs) and hemoglobin distribution width (HDW) were higher in the MDS group than in the other cytopenia group and showed the greatest difference between both groups, with an area under the curve (AUC) of 0.766 (0.678-0.855) and 0.786 (0.702-0.870), respectively. The platelet distribution width was higher in the MDS group than in the other cytopenia group, with an AUC of 0.697 (0.623-0.770). WBC CPD extracted from histograms, especially Atyp-PMN-loc and Neu-ALL-M, showed high AUCs of 0.815 (0.750-0.879) and 0.778 (0.711-0.845), respectively. Our findings demonstrate the clinical utility of CPD and hematology parameters of the Abbott Alinity hq analyzer in the differential diagnosis of MDS.

Usability and Performance Testing of Pointcheck™: A Noninvasive Neutropenia Screening Device for Chemotherapy Outpatients

Usability and Performance Testing of Pointcheck™: A Noninvasive Neutropenia Screening Device for Chemotherapy Outpatients

Neonatal Reference Intervals for the Complete Blood Count Parameters MicroR and HYPO-He: Sensitivity Beyond the Red Cell Indices for Identifying Microcytic and Hypochromic Disorders

To create neonatal reference intervals for the MicroR and HYPO-He complete blood count (CBC) parameters and to test whether these parameters are sensitive early markers of disease at early stages of microcytic/hypochromic disorders while the CBC indices are still normal. We retrospectively collected the CBC parameters MicroR and HYPO-He, along with the standard CBC parameters, from infants aged 0-90days at Intermountain Healthcare hospitals using Sysmex hematology analyzers. We created reference intervals for these parameters by excluding values from neonates with proven microcytic disorders (ie, iron deficiency or alpha thalassemia) from the dataset. From >11 000 CBCs analyzed, we created reference intervals for MicroR and HYPO-He in neonates aged 0-90days. The upper intervals are considerably higher in neonates than in adults, validating increased anisocytosis and polychromasia among neonates. Overall, 52% of neonates with iron deficiency (defined by reticulocyte hemoglobin equivalent <25 pg) had a MicroR >90% upper interval (relative risk, 4.14; 95% CI, 3.80-4.53; P<.001), and 68% had an HYPO-He >90% upper interval (relative risk, 6.64; 95% CI, 6.03-7.32; P<.001). These 2 new parameters were more sensitive than the red blood cell (RBC) indices (P<.001) in identifying 24 neonates with iron deficiency at birth. We created neonatal reference intervals for MicroR and HYPO-He. Although Sysmex currently designates these as research use only in the US, they can be measured as part of a neonate's CBC with no additional phlebotomy volume or run time and can identify microcytic and hypochromic disorders even when the RBC indices are normal.

Red blood cell distribution width as a novel prognostic marker after myocardial revascularization or cardiac valve surgery

The red blood cell distribution width (RDW) measures the variability in the size of circulating erythrocytes. Previous studies suggested a powerful correlation between RDW obtained from a standard complete blood count and cardiovascular diseases in both primary and secondary cardiovascular prevention. The current study aimed to evaluate the prognostic role of RDW in patients undergoing cardiac rehabilitation after myocardial revascularization and/or cardiac valve surgery. The study included 1.031 patients with available RDW levels, prospectively followed for a mean of 4.5 ± 3.5 years. The mean age was 68 ± 12 years, the mean RDW was 14.7 ± 1.8%; 492 patients (48%) underwent cardiac rehabilitation after myocardial revascularization, 371 (36%) after cardiac valve surgery, 102 (10%) after valve-plus-coronary artery by-pass graft surgery, 66 (6%) for other indications. Kaplan–Meier analysis and Cox hazard analysis were used to associate RDW with mortality. Kaplan–Meier analysis demonstrated worse survival curves free from overall (log-rank p < 0.0001) and cardiovascular (log-rank p < 0.0001) mortality in the highest RDW tertile. Cox analysis showed RDW levels correlated significantly with the probability of overall (HR 1.26; 95% CI 1.19–1.32; p < 0.001) and cardiovascular (HR 1.31; 95% CI 1.23–1.40; p < 0.001) mortality. After multiple adjustments for cardiovascular risk factors, hemoglobin, hematocrit, C-reactive protein, microalbuminuria, atrial fibrillation, glomerular filtration rate,left ventricular ejection fraction and number of exercise training sessions attended, the increased risk of overall (HR 1.10; 95% CI 1.01–1.27; p = 0.039) and cardiovascular (HR 1.13; 95% CI 1.01–1.34; p = 0.036)mortality with increasing RDW values remained significant. The RDW represents an independent predictor of overall and cardiovascular mortality in secondary cardiovascular prevention patients undergoing cardiac rehabilitation.

Computer Vision and Deep Learning Assisted Microchip Electrophoresis for Integrated Anemia and Sickle Cell Disease Screening

Introduction: Anemia affects a third of the world's population with the heaviest burden borne by women and children. Anemia leads to preventable impaired development in children, as well as high morbidity and early mortality among sufferers. Inherited hemoglobin (Hb) disorders, such as sickle cell disease (SCD), are associated with chronic hemolytic anemia causing high morbidity and mortality. Anemia and SCD are inherently associated and are both prevalent in the same regions of the world including sub-Saharan Africa, India, and south-east Asia. Anemia and SCD-related complications can be mitigated by screening, early diagnosis followed by timely intervention. Anemia treatment depends on the accurate characterization of the cause, such as inherited Hb disorders. Meanwhile, Hb disorders or SCD treatments, such as hydroxyurea therapy, requires close monitoring of blood Hb level and the patient's anemia status over time. As a result, it is crucially important to perform integrated detection and monitoring of blood Hb level, anemia status, and Hb variants, especially in areas where anemia and inherited Hb disorders are the most prevalent. Blood Hb level (in g/dL) is used as the main indicator of anemia, while the presence of Hb variants (e.g., sickle Hb or HbS) in blood is the primary indicator of an inherited disorder. The current clinical standards for anemia testing and Hb variant identification are complete blood count (CBC) and High-Performance Liquid Chromatography (HPLC), respectively. State-of-the-art laboratory infrastructure and trained personnel are required for these laboratory tests. However, these resources are typically scarce in low- and middle-income countries, where anemia and Hb disorders are the most prevalent. As a result, there is a dire need for high accuracy portable point-of-care (POC) devices to perform integrated anemia and Hb variant tests with affordable cost and high throughput. Methods: In 2019, the World Health Organization (WHO) listed Hb electrophoresis as an essential in vitro diagnostic (IVD) technology for diagnosing SCD and sickle cell trait. We have leveraged the common Hb electrophoresis method and developed a POC microchip electrophoresis test, Hemoglobin Variant/Anemia (HbVA). This technology is being commercialized under the product name "Gazelle" by Hemex Health Inc. for Hb variant identification with integrated anemia detection (Fig. 1A&amp;B). We hypothesized that computer vision and deep learning will enhance the accuracy and reproducibility of blood Hb level prediction and anemia detection in cellulose acetate based Hb electrophoresis, which is a clinical standard test for Hb variant screening and diagnosis worldwide (Fig. 1C). To test this hypothesis, we integrated, for the first time, a new, computer vision and artificial neural network (ANN) based deep learning imaging and data analysis algorithm, to Hb electrophoresis. Here, we show the feasibility of this new, computer vision and deep learning enabled diagnostic approach via testing of 46 subjects, including individuals with anemia and homozygous (HbSS) or heterozygous (HbSC or Sβ-thalassemia) SCD. Results and Discussion: HbVA computer vision tracked the electrophoresis process real-time and the deep learning neural network algorithm determined Hb levels which demonstrated significant correlation with a Pearson Correlation Coefficient of 0.95 compared to the results of reference standard CBC (Fig.1D). Furthermore, HbVA demonstrated high reproducibly with a mean absolute error of 0.55 g/dL and a bias of -0.10 g/dL (95% limits of agreement: 1.5 g/dL) according to Bland-Altman analysis (Fig. 1E). Anemia determination was achieved with 100% sensitivity and 92.3% specificity with a receiver operating characteristic area under the curve (AUC) of 0.99 (Fig. 1F). Within the same test, subjects with SCD were identified with 100% sensitivity and specificity (Fig. 1G). Overall, the results suggested that computer vision and deep learning methods can be used to extract new information from Hb electrophoresis, enabling, for the first time, reproducible, accurate, and integrated blood Hb level prediction, anemia detection, and Hb variant identification in a single affordable test at the POC. Disclosures An: Hemex Health, Inc.: Patents &amp; Royalties. Hasan:Hemex Health, Inc.: Patents &amp; Royalties. Ahuja:Genentech: Consultancy; Sanofi-Genzyme: Consultancy; XaTec Inc.: Consultancy; XaTec Inc.: Research Funding; XaTec Inc.: Divested equity in a private or publicly-traded company in the past 24 months; Genentech: Honoraria; Sanofi-Genzyme: Honoraria. Little:GBT: Research Funding; Bluebird Bio: Research Funding; BioChip Labs: Patents &amp; Royalties: SCD Biochip (patent, no royalties); Hemex Health, Inc.: Patents &amp; Royalties: Microfluidic electropheresis (patent, no royalties); NHLBI: Research Funding; GBT: Membership on an entity's Board of Directors or advisory committees. Gurkan:Hemex Health, Inc.: Consultancy, Current Employment, Patents &amp; Royalties, Research Funding; BioChip Labs: Patents &amp; Royalties; Xatek Inc.: Patents &amp; Royalties; Dx Now Inc.: Patents &amp; Royalties.

Salivary biochemical variables in liver transplanted children and young adults.

To investigate associations between levels of blood parameters used to monitor liver-transplanted children with their salivary levels, and compare the salivary parameters of transplant recipients with those of healthy controls. Saliva and blood samples from 30 liver transplanted recipients, mean age 11.7years and saliva from age and sex matched 27 healthy patients were analyzed using a standard complete blood count test. Uric acid and alkaline phosphatase levels correlated significantly between saliva and blood samples in the transplanted subjects. Median salivary sodium level was significantly lower and the median salivary potassium level significantly higher in transplant recipients compared with healthy subjects. No differences were found between the groups in salivary glucose, urea, chloride, total protein, albumin, calcium, phosphorus, uric acid, total bilirubin, alkaline phosphatase, lactate dehydrogenase (LDH), glutamic oxaloacetic transaminase (GOT), triglycerides, cholesterol, iron, transferrin, glutamic pyruvic transaminase (GPT) and gamma-glutamyltranspeptidase (GGT). Specific correlations of serum and salivary chemistry were found in liver transplant patients. Such information may lead to the development of noninvasive monitoring tools for this population.

Vitamin B12 Deficiency Among the Healthy Jordanian Adult Population: Diagnostic Levels, Symptomology and Risk Factors.

Compared to the data available for developed countries, there is a marked scarcity of information on the levels and symptomology of vitamin B12 deficiency in developing countries, particularly in the Middle Eastern region. To explore (a) the risk factors associated with a deficiency of vitamin B12, and (b) the baseline (cut-off) serum level of vitamin B12 for a clinically-symptomatic deficiency in the Jordanian adult population. A total of 485 subjects were included in this study. Blood samples were drawn for biochemical analysis and data regarding socio-demographics, general health, anthropometric measures, and past medical, surgical, and medication history were collected. To explore the cut-off point, we compared all parameters included in a standard complete blood count as well as the main symptoms reported to be associated with B12 deficiency between groups of different B12 cut-off values, consisting of those above and below 200, 175, 150 and 125 pg/ml. Low dairy intake habits, age, recurrent headaches, heartburn, and peptic ulcer disease were found to be significantly associated with lower vitamin B12 levels. Surprisingly, daily smoking was associated with significantly higher B12 levels. The results revealed that none of the included potential indicators of B12 deficiency could be considered an indicative feature of deficiency. There were no significant differences neither in the symptoms nor in the CBC parameters between any of the tested study groups. Low dairy intake, older ages, recurrent headaches, heartburn, and peptic ulcer disease all could be considered as risk factors of having low vitamin B12 levels within the Jordanians. Also, they tend to have lower levels of vitamin B12 levels, in comparison to countries in the West, without necessarily having deficiency symptoms. The cut-off value to diagnose functional B12 deficiency could be less than 125 pg/ml for the Jordanians. More local studies are needed to establish an accurate vitamin B12 cut-off value for the population in Jordan.

Impact of relacorilant, a selective glucocorticoid receptor antagonist, on the immunosuppressive effects of endogenous cortisol.

3091 Background: Cortisol, an endogenous glucocorticoid receptor (GR) agonist, controls a broad transcriptional program that affects T-cell activation, pro-inflammatory cytokine secretion, and immune cell trafficking. By selectively antagonizing GR, relacorilant may reverse the immunosuppressive effects of cortisol in solid tumor cancers. Methods: Immune cells and GR expression were assessed by IHC and calculated based on The Cancer Genome Atlas (TCGA) data. Human PBMCs were stimulated with αCD3+IL-12 +/- cortisol or cortisol + relacorilant. EG7 tumor-bearing mice were treated with anti-PD1 (RMP1-14) ip Q5D +/- relacorilant QD. Whole blood mRNA was measured via Nanostring, hematology was performed using standard complete blood count assays, and cytokines were assessed by immunoassays in study NCT02762981. Results: GR expression was observed in human tumor and immune cells. Its abundance was positively correlated with tumor infiltration of TH2, Treg, and PDL1+ cells ( P&lt; .001) and negatively correlated with TH1 cells ( P&lt; .001). In PBMCs, cortisol inhibited, and relacorilant restored, CD8+ T-cell activation ( P&lt; .001) and pro-inflammatory cytokine secretion (TNFα P= .006, IFNγ P&lt; .05). In the EG7 syngeneic model, relacorilant increased αPD1 efficacy ( P= .007) and decreased circulating IL-10 ( P&lt; .002). In patients with advanced solid tumors, relacorilant + nab-paclitaxel systemically suppressed the expression of canonical GR-controlled genes ( ptgs2 P&lt; .001) and genes encoding candidate-immunomodulatory drug targets ( cxcl8, ptger4, ido1; P&lt; .001). In a small subset of patients (n = 11), sustained clinical response was associated with increased T-cell count ( P= .06) and IFNγ ( P= .03), as well as decreased Tregs ( P= .06) and IL-10 ( P= .03). Conclusions: Evidence of T-cell activation by relacorilant was observed in PBMCs, syngeneic mouse tumors, and patients with sustained response in a Phase 1 study. This supports the hypothesis that relacorilant can reverse immune suppression by endogenous cortisol in solid tumor cancers. Clinical studies with immune checkpoint inhibitors and relacorilant are planned.