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Midwest Clinical and Translational Research Meeting of CSCTR And Abstracts J Investig Med: first published as 10.1136/jim-2020-MW on 11 June 2020. Downloaded from Bio-engineering Results Our results showed that individuals with homozygous SCD (HbSS) had significantly higher WBV compared to healthy controls when the hematocrit of the samples were A01 MICRO PARTICLE IMAGE VELOCIMETRY FOR IN VITRO adjusted to 50% (4.8±0.41 cP vs 4.15±0.07 cP, p<0.05, ASSESSMENT OF PATIENT SPECIFIC WHOLE BLOOD Mann-Whiney U-test). However, HbSS SCD usually leads to RHEOLOGY significant anemia and lower hematocrit, which may lead to a 1Erdem Kucukal, 2Yuncheng Man, 3Ran An, 4Jane A Little, 2Umut A Gurkan. 1Case Western lower WBV compared to the non-SCD population. Interest- Reserve University, Cleveland Heights, OH; 2Case Western Reserve University, OH; 3Case ingly, HbSS SCD samples displayed significantly lower WBV Western Reserve University, Cleveland, OH; 4University of North Carolina, NC compared to the healthy controls when unprocessed whole blood samples were tested (3.8±0.67 cP vs 4.49±0.32 cP, 10.1136/jim-2020-MW.1 p<0.05, Mann-Whitney U-test). We observed a significant het- erogeneity in terms of WBV among the HbSS subjects, where Introduction/Background Whole blood visocity (WBV) is a the WBV values ranged between 3.12 cP and 5.82 cP. Nota- pivotal biophysical parameter in many cardiovascular diseases bly, subjects with a recent blood transfusion (<3 months, and hematological disorders such as sickle cell disease (SCD). n=9) had a significantly higher WBV compared to those who It can be influenced by a variety of factors including plasma were on hydroxyurea (HU) (n=15; 4.01±0.71 cP vs 3.74 viscosity, RBC deformability, hematocrit, and plasma protein ±0.71 cP, p<0.05, Mann-Whitney U-test). Subjects who were levels. Although the separate effects of these factors on WBV both on HU and with recent transfusion history were determination have been well documented, a more compre- excluded from this analysis. hensive approach that takes into account all these factors in RBC adhesion to laminin (LN) has been repeatedly shown a patient-specific fashion is needed to better understand the to correlate with disease severity in SCD. Since our results role of WBV in SCD pathophysiology. Here, we describe a here also suggest a significantly heterogeneous WBV profile microfluidic platform integrated with the micro particle among the study group, we next sought to determine whether image velocimetry (PIV) technique to quantify WBV using an association between RBC adhesion and WBV existed. unprocessed whole blood samples from individuals with Because the lowest WBV of HbAA samples that we tested was SCD. Further, for the first time, we report both WBV and approximately 4 cP, we segregated the SCD study population RBC adhesion levels simultaneously using this microfluidic into two groups: lower WBV group (<4 cP, N=8) and higher system. WBV (>4 cP, N=21). Interestingly, subjects with a lower Objective(s) To assess whole blood viscosity (WBV) of subjects WBV exhibited significantly greater RBC adhesion to LN com- with sickle cell disease (SCD) using a high-throughput micro- pared to those with a higher WBV (mean adhesion ± SD: fluidic system integrated with a micro particle image velocime- 957±767 per fov vs 452±331 per fov, p<0.05, Student’s t- try (PIV) technique under normoxic or hypoxic conditions, test, fov: field of view). These findings suggest that WBV and and to examine its clinical associations and impact. RBC adhesion may play distinct roles in the pathophysiology Methods Blood collection – Whole blood samples from de- of homozygous SCD. identified healthy donors and SCD subjects ( 18) were col- Hypoxia plays a crucial role in SCD pathophysiology as it http://jim.bmj.com/ lected in EDTA (Ethylenediaminetetraacetic acid) containing leads to the formation of abnormally adhesive and stiff red vacutainers based on an Institution Review Board (IRB) blood cells due to hemoglobin polymerization. Consequently, approved protocol. Subject clinical information, including we integrated a micro gas exchanger to our microfluidic sys- medical treatments and previous comorbidities, were acquired tem, as we have previously described, in order to probe the after patients had provided a written consent from the Adult change in WBV as well as RBC adhesion to LN from nor- Sickle Cell Clinic at University Hospitals Cleveland Medical moxic to hypoxic conditions. The hypoxic viscosity results Center (UHCMC) in Cleveland, Ohio. A total number of 10 on September 26, 2021 by guest. Protected copyright. showed that WBV of control blood samples (HbAA, N=3) samples from healthy subjects (HbAA), 14 samples from sub- remained relatively unchanged, while the HbSS samples jects with HbSC, and 29 samples from subjects with HbSS (N=10) became more viscous under hypoxia (p=0.007, stu- were tested. dent’s t-test). More interestingly, we observed an inverse rela- Micro PIV for velocity measurements – Whole blood sam- tionship between hypoxic WBV and RBC adhesion to LN in ples were injected into rectangular microfluidic devices (4 mm hypoxia, a similar behavior as in normoxic conditions (Pear- x 0.05 mm) under constant physiological pressure (20 mm. son’s correlation coefficient = -0.6, p=0.03, one-way Hg), and high resolution images of the blood flow were ANOVA). In other words, RBCs from samples with a lower acquired via an inverted microscope (Olympus IX83). The hypoxic WBV had an increased propensity to adhere to LN in images were then cross-correlated to compute the mean flow hypoxia. velocities in a given region of interest, which was a function Conclusions The presented microfluidic system demonstrates of fluid viscosity. The conversion from mean blood velocity to a new approach for simultaneous analysis of RBC adhesion microfluidic WBV was carried out using a standard curve gen- and WBV in SCD. The results of this study suggest that erated by using clinically measured viscosity values (piston WBV is significantly lower in those with HbSS SCD, likely style viscometer). due to severe anemic conditions. Further, WBV inversely RBC adhesion assay – The adhesion tests were performed associates with RBC adhesion both in normoxic and hypoxic in laminin-functionalized microfluidic channels. The blood conditions, which has a subject-specific profile. Future studies samples with a volume of 15 ml were first injected into the will aim to tease out the individual as well as synergistic microchannels at a shear stress of 1 dyne/cm2, and then non- contributions of WBV and RBC adhesion to clinical manifes- adherent cells were removed by flowing a buffer solution (1% tations of SCD. BSA in PBS) at the same shear stress. 1026 J Investig Med 2020;68:1026–1124 Abstracts J Investig Med: first published as 10.1136/jim-2020-MW on 11 June 2020. Downloaded from Bone Marrow Transplant B17 NO ASSOCIATION OF TELOMERE LENGTH WITH FRAILTY IN PRE-HEMATOPOIETIC CELL TRANSPLANT POPULATION 1Hok Sreng Te, 2Troy Lund, 2Bharat Thyagarajan, 2Todd DeFor, 2Mukta Arora. 1University of Minnesota, Minneapolis, MN; 2University of Minnesota 10.1136/jim-2020-MW.2 Introduction/Background Frailty affects about 10% of commun- ity dwelling elderly ( 65 years) and is defined as a state of diminished physiological reserve and increased vulnerability to stress associated with aging and decline in function across multi- ple physiological systems. The hematopoietic cell transplant (HCT) population is unique in that they have already suffered Abstract B17 Figure 1 stresses to the hematopoietic cells imposed by prior chemothera- Proportion of patients with frailty score of 0 (not-frail), 1–2 (pre-frail) peutic treatments (direct cytotoxicity, DNA damage, and replica- and 3+ (frail) across each group of telomere length and age group tive, inflammatory or oxidative stress), which may result in altered leukocyte telomere dynamics even before HCT. These unintentional weight loss (58% versus 42%), weakness (53% stressors are also known physiological correlates of aging and versus 47%), and low energy expenditure (52% versus 48%), may also impact frailty. We hence evaluated the prevalence of but this did not attain statistical significance. frailty and its impact on survival and the predictive capability of Conclusions Independent of comorbidities and age, pre-HCT mean leukocyte telomere length on frailty in a HCT population. frailty was noted in 21% of our HCT population. A higher Objective(s) The objective of this study is to determine the prev- mortality at 1-year post HCT was noted in frail HCT recipi- alence of pre-HCT frailty, the impact of pre-HCT frailty on ents, indicating the need to monitor these patients closely and overall survival at 1-year post-HCT, and the association between plan personalized interventions in these patients. We did not pre-HCT frailty and pre-HCT recipient telomere length. find an association between frailty and telomere length. Future Methods We conducted a prospective, longitudinal study of studies need to evaluate underlying pathophysiology in frail 117 patients undergoing HCT at age 40 years in 2014 and patients to help identify mechanisms of frailty. 2015 at the University of Minnesota. Pre-HCT frailty pheno- type was constructed based on patient performance to the fol- lowing criteria: unintentional weight loss, self-reported exhaustion, weakness, slow walking speed, and low physical http://jim.bmj.com/ activity. Frailty was defined as a clinical syndrome meeting 3 Cardiology/Cardiovascular Disease of the criteria, while pre-frailty was defined as meeting 1 or 2 of the criteria. The study population was analyzed in 2 B21 CARDIAC HEMODYNAMIC AND REMODELING groups: 40–59 years and 60 years. Average telomere length PATTERNS FOLLOWING TRANSCATHETER AORTIC was estimated by Southern Blot analysis. Recipients were VALVE REPLACEMENT grouped by age and quartiles of telomere length into four 1Julien Feghaly, 2Zachary Oman, 2Debapria Das, 2Steven Smart.
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