Reticulocyte Indices Research Articles

Prognostic value of reticulocyte production ability in patients with chronic heart failure

Abstract Background Anaemia is frequently observed in patients with chronic heart failure (CHF), and many studies have elucidated the association between anaemia and mortality in patients with CHF. The quality of erythropoiesis is an intrinsic aspect of pathophysiology of anaemia, and the ability of reticulocyte production would be one of the pivotal factors in the quality of erythropoiesis. Purpose We sought to investigate whether the reticulocyte production ability is associated with clinical outcomes, and clarify the pathophysiology of reticulocyte production in patients with CHF. Methods We prospectively examined 1,328 symptomatic CHF patients (median age 71 [IQR 65-82] years, mean left ventricular ejection fraction 50 ± 16%, median NT-proBNP 743 [IQR 360–1710] pg/mL) in a multicentre registry with 27 Japanese sites between January 2020 and October 2023. The reticulocyte production ability was evaluated by calculating the reticulocyte production index (RPI) using reticulocyte counts and serum haematocrits. Patients were divided into four groups according to the presence or absence of WHO defined anaemia and the median value of RPI. The primary outcome was a composite of all-cause death and hospitalisation for worsening HF. Results During a median follow-up period of 551 (IQR 321–712) days, the primary outcome occurred in 219 (17%) patients. The group with anaemia and high RPI (≥0.978, the median) showed the highest incidence of the primary outcome among the groups (P < 0.001) (Figure 1). Multivariable regressions showed that higher RPI was independently associated with the higher risk of the primary outcome, even after adjustment for NT-proBNP, sodium, haemoglobin, serum iron, erythropoietin, and MAGGIC risk score (HR 1.41, 95% CI 1.09–1.82). Erythrocyte counts were significantly increased in patients with higher RPI compared to those with lower RPI in the groups without anaemia (P < 0.001)whereas there were no significant differences on erythrocyte counts between the groups with anaemia (P = 0.90) (Figure 2A). Patients with higher RPI had higher log erythropoietin levels compared to those with lower RPI in either anaemic or non-anaemic group (Figure 2B). In addition, serum iron levels and transferrin saturation did not significantly differ between the RPI groups in either anaemic or non-anaemic group (Figures 2C and D). Furthermore, in patients without anaemia, erythrocyte counts positively correlated with RPI (R = 0.27, P < 0.001) whereas there was very weak correlation between erythrocyte counts and RPI in those with anaemia (R = 0.07, P = 0.045). Conclusions Higher RPI was independently associated with worse clinical outcomes, suggesting RPI could be a surrogate marker for risk stratification in patients with CHF, especially in anaemic population. Higher reticulocyte production ability might reflect the impaired maturation of reticulocytes to erythrocytes independent of iron parameters.

Fatty Acid β-Oxidation May Be Associated with the Erythropoietin Resistance Index in Stable Patients Undergoing Haemodialysis.

Lipid metabolism and adiponectin modulate erythropoiesis in vitro and in general population studies and may also affect responsiveness to erythropoietin in patients undergoing haemodialysis (HD). However, little is known about the impact of lipid-associated biomarkers on reticulocyte production and erythropoietin resistance index (ERI) in patients undergoing HD. Therefore, we aimed to investigate their impacts in 167 stable patients undergoing HD. Pre-dialysis blood samples were collected and analysed for reticulocyte counts and serum lipid profiles by routine analyses and serum carnitine profiles (C0-C18) by LC-MS/MS. ERI was calculated as erythropoietin dose/kg/week normalized for haemoglobin levels. The independent positive determinants of reticulocyte count were log [Triglyceride (TG)] and logC18:1. A large proportion of longer-chain acylcarnitines was positively correlated with reticulocyte counts, possibly resulting from the accumulation of acylcarnitines in mitochondria undergoing fateful exocytosis from reticulocytes. These results indicate a possible association between reticulocyte formation and reduced β-oxidation, which occurs during the peripheral phase of erythroblast enucleation. Total cholesterol (TC) and log [C2/(C16 + C18:1)] as a putative marker of β-oxidation efficiency were negative independent determinants of ERI. Moreover, acyl chain length had a significantly positive impact on the correlation coefficients of individual acylcarnitines with ERI, suggesting that enhanced β-oxidation may be associated with reduced ERI. Finally, adiponectin had no independent association with reticulocyte counts or ERI despite its negative association with HDL-C levels. Enhanced fatty acid β-oxidation and higher TC levels may be associated with lower ERI, whereas higher TG levels and longer acylcarnitines may be related to the latest production of reticulocytes in stable patients undergoing HD.

Red cell size factor is a sensitive index in the early diagnosis of nondigestive tract cancer-related anemia: An observational study.

Cancer-related anemia (CRA) is a common comorbidity in cancer patients, and it can lead to a worse prognosis. The aim of this cross-sectional study is to investigate the clinical value of the red cell size factor (Rsf) in the early diagnosis of nondigestive tract CRA. A total of 231 patients with nondigestive tract solid cancer were included, and they were divided into anemic and nonanemic subjects according to the hemoglobin (Hb) levels. A BC-7500 blood analyzer was used to detect the indices of red blood cell and reticulocyte, and the mean corpuscular volume (MCV), mean reticulocyte volume (MRV), reticulocyte hemoglobin (RHE) content, and reticulocyte production index were observed. Subsequently, the Rsf was calculated. Receiver operating characteristic curve analysis was used to evaluate the identifying power of Rsf for anemia diagnosed by the combination of RHE and reticulocyte production index. The adjusted-multivariate analysis and quartiles were used to assess the relation of reduced Rsf level with the risk and incidence of anemia diagnosed by combining the MCV, MCH, and mean corpuscular hemoglobin concentration (MCHC), respectively. Rsf levels showed no statistical differences between anemia and nonanemia subjects grouped by Hb (P > .05). Rsf has a high correlation with the RHE level (R > 0.900, P < .001), or MCV, mean corpuscular hemoglobin (MCH), and MCHC in anemia patients (r: 0.435-0.802, P < .001). Receiver operating characteristic curves showed that Rsf had the highest overall area under curve of 0.886 (95% confidence interval: 0.845-0.927) in identifying anemia of cancer patients (P < .001). When the optimal cutoff values of Rsf were set at 97.05 fl in males and 94.95 fl in females, the sensitivity and specificity were 0.94 and 0.76, and 0.98 and 0.75, respectively. Being treated as a categorical variable, Rsf had a highest odds ratio value of 30.626 (12.552-74.726; P < .001) for the risk of anemia. The increment of Rsf quartiles was highly associated with the decreased incidence of overall anemia (P trend < 0.001). The study suggests that decreased Rsf level is a potentially powerful predictor of overt anemia in nondigestive tract cancer, and it can be used as a convenient, practical, cost-free, and sensitive index in early diagnosis of nondigestive tract CRA.

Validation of the Sysmex XN-V hematology analyzer for feline specimens.

The Sysmex XN-V is derived from the new Sysmex XN series of human hematology analyzers. The main changes from the previously validated XT-2000iV analyzer include an optic-fluorescent analysis for platelets and a nucleated red blood cell (NRBC) count. We aimed to validate the Sysmex XN-V for feline blood following the American College for Veterinary Clinical Pathology and International Council for Standardization in Hematology recommendations. Feline EDTA blood specimens were analyzed on the Sysmex XN-V to evaluate repeatability, linearity, comparison with the XT-2000iV analyzer and manual methods, stability, and to verify the previously established Sysmex XT-2000iV RIs. Repeatability was excellent for most variables. Visually determined linearity was excellent or good for most variables except eosinophils and platelet variables. The correlation between the XN-V and XT-2000iV analyzers was good (≥0.82) for all variables except reticulocyte indices. Correlations between the Sysmex XN-V and manual differential counts were good to excellent for most variables, acceptable for neutrophils, and fair for monocytes and NRBC. The previously established Sysmex XT-2000iV RIs can be used to interpret results from the Sysmex XN-V analyzer for most variables except red cell distribution width and reticulocyte variables. The RI for platelet variables could not be evaluated because of platelet clumps. Changes in the Sysmex XN-V measurements after storage at 4 and 24°C were similar to those described for the Sysmex XT-2000iV analyzer. The performance of the Sysmex XN-V analyzer was good and compared favorably with the Sysmex XT-2000iV analyzer.

#3090 Parvovirus infection in renal transplant recipients: a systematic review of clinical profile and treatment strategies

Abstract Background and Aims Parvovirus B 19 (PVB-19) virus infection can cause severe, persistent and/or refractory anaemia in kidney transplant recipients (KTR). There is paucity of data on the clinical manifestations, diagnosis and management of this infection and limited to case reports and case series. Method We performed a literature search of kidney transplant recipients infected with PVB-19 in MEDLINE, EMBASE and Cochrane databases and 93 articles were identified as suitable for assessment. The review was conducted in accordance with PRISMA statement. Results A total of two hundred and eighty-two patients were included from the studies assessed in the review. The mean age of the cohort was 37.05 years. The infection was more common in males when compared to females. The median onset of PVB-19 infection post transplantation was at 5 weeks (2, 15 weeks). One fourth of the recipients had at least one episode of rejection prior to PVB-19 infection. Majority of the patients were on Tacrolimus, MMF and corticosteroids maintenance. One third of the patients had co-infections with other organisms (EBV, CMV, etc.). Anemia and fever were the most common manifestations among all the kidney transplant patients infected with PVB-19 while CNS manifestations and rash were least reported. Hemoglobin was low (&amp;lt;10 mg/dl) in ninety eight percent of the patients (n=151). The lowest hemoglobin reported in the due course of illness was 6 mg/dl (5, 7.2 mg/dl) and with corrected reticulocyte count being only 0.2% (0.1%, 0.6%). The most consistent finding on peripheral smear was normocytic, normochromic anemia while a few reported helmet cells too. Pure Erythroid hypoplasia alone (45.3%) combined with giant pronormoblasts and inclusion bodies (49%) were the most common findings on bone marrow aspirate. Majority of PVB-19 diagnosis was made with the help of multiple methods of detection like IgM assay, PCR—qualitative and quantitative (42.2%). The average PCR copies detected were 1.3*107 (4.28*105, 2.53*109). IVIG only (47.2%) and combined IVIG + immunosuppression reduction (41.2%) was the mainstay of the treatment in majority. The median time to recover was 8 weeks (3, 16 weeks). One fourth of the patients had recurrence after recovery. A majority of the patients (96.7%) recovered from reactivation of PVB-19. Conclusion Parvovirus infection is not uncommon in the early post-transplant period. Though data is insufficient, immunosuppression reduction with IVIG remains the treatment of choice for affected patients. Prospective studies to assess the hemoglobin cutoffs and dose of IVIG are needed to frame screening and treatment protocols.

Evaluation of erythrocyte and reticulocyte indices in a newborn with severe hemolytic disease

Currently, significant progress has been made in the prevention, diagnosis and treatment of hemolytic disease of the fetus and newborn. However, the development of anemia in a newborn child due to this disease remains an urgent problem for both neonatologists and pediatricians. In such children, a comprehensive assessment of the hemogram is of particular importance. The study of erythrocyte and reticulocyte parameters of a general blood test is necessary to determine prognostic criteria for the restoration of hematopoiesis and determine the status of iron deficiency. A clinical observation of a newborn child with the development of anemia due to hemolytic disease of the fetus and newborn is presented (clinical case). A dynamic analysis of reticulocyte parameters (absolute and relative numbers) and reticulocyte fractions was carried out. An increase in reticulocytes (absolute and relative numbers) was established, mainly due to the fraction of immature reticulocytes during the development of anemia in a newborn child. Such indicators as the hemoglobin content in reticulocytes, the average hemoglobin content in an erythrocyte, the average hemoglobin concentration in an erythrocyte in a general blood test in an infant during the neonatal period remained within the reference values. The data obtained made it possible to determine a personalized approach to the treatment of anemia and avoid blood transfusion in this child.

Evaluation of reticulocyte indices and mathematical formulas for beta-thalassemia carriers screening among Egyptian beta-thalassemia patients’ families

Background Egypt lies within the global thalassemia belt, where beta-thalassemia presents a notable socioeconomic burden. Patients and methods One hundred fifty-seven relatives of beta-thalassemia patients were included in this cross-sectional study. It aimed to ascertain a cost-efficient and accurate screening approach for identifying beta-thalassemia carriers (BTC) by evaluating some erythrocyte and reticulocyte parameters measured on Advia 2120 analyzer (Siemens) and assessing four mathematical Formulas (Metzner index, Green and King et al., Roth et al., and Vicinanza et al. formulas). Results Participants were stratified into distinct categories: BTC (42%), normal (24.2%), mild iron-deficiency anemia (IDA) (17.2%), severe IDA (3.8%), and a subset with both IDA and BTC (mix group) accounting for 12.7%. Receiver operating characteristic curve analysis identified absolute reticulocyte count as the optimal discriminator (area under a curve=0.837) between BTC and IDA. The Mentzer index and Green and King et al. formula demonstrated superior diagnostic utility among the evaluated formulas. However, none of the formulas effectively distinguished between BTC and the mix group, a frequently overlooked and challenging group for diagnosis. Interestingly, our data highlighted absolute reticulocyte count and percentage as the sole parameters capable of distinguishing BTC from the mix group. Consequently, two algorithms were developed. The first one had a good agreement (κ=0.62) with 77.3% concordance, while the second one demonstrated improved screening performance (κ=0.66) with 79.8% agreement. Conclusion The optimal screening strategy involves distinguishing between various categories of both IDA and BTC. Our algorithms, designed as a low-cost and user-friendly mass screening tool, may serve this purpose effectively.

Reticulocyte Indexes and their Significance

A crucial diagnostic tool for assessing, categorizing, and monitoring the effectiveness of anemia treatment is the reticulocyte count. The quality of erythropoiesis is an intrinsic aspect of the pathophysiology of anemia, and the ability of reticulocyte production would be one of the pivotal factors in the quality of erythropoiesis. However, in this minireview, we explain clearly whether the reticulocyte index’s ability is associated with clinical outcomes. There are several ways to describe the reticulocyte count: as a proportion of all red blood cells, as an absolute count, as a corrected count, as the Reticulocyte Production Index (RPI), or as the Immature Reticulocyte Fraction (IRF). Reticulocyte indexes aid in determining the etiology of anemia. They can distinguish between anemia-related causes and other dyserythropoietic problems.

Reticulocytes and Related Parameters in Clinical Practice

The development of hematologic analyzers allowed us to assess several additional parameters derived in addition to the number of reticulocytes. There is, among others, the calculation of immature reticulocyte fractions, reticulocyte maturity index, reticulocyte production index, and equivalent reticulocyte hemoglobin. The advantage of the equivalent hemoglobin and the content of hemoglobin is the insensitivity to interference by inflammation. These newly introduced parameters expand the possibilities to access bone marrow erythropoietic activity, iron stores, and the effectiveness of iron supplementation, vitamin B12, and folic acid supplementation. These parameters are also used to make decisions about blood transfusions or to detect occult internal bleeding.

The mean reticulocyte volume is a valuable index in early diagnosis of cancer-related anemia.

Cancer-related anemia (CRA) is a functional iron deficient anemia, and the early diagnosis will improve the prognosis of the patients. This prospective study aimed to investigate the utility of mean reticulocyte volume (MRV) in the early diagnosis of CRA. A total of 284 first-diagnosed cancer patients were enrolled, and the subjects were assigned anemia and non-anemia groups by hemoglobin (Hb) concentrations. The mature RBC and reticulocyte indices were detected with BC-7500 blood analyzer, and the MRV, reticulocyte hemoglobin (RHE) content, and reticulocyte production index (RPI) were obtained. ROC curves were constructed in identifying anemia diagnosed by the combination of RHE and RPI. An adjusted multivariate analyse and quartiles were used to assess the associations of MRV with early CRA diagnosed by combining RBC indices (MCV, MCH and MCHC), respectively. No statistical differences were observed in MCV, RHE and MRV levels between anemia and non-anemia subjects (p>0.05). MRV exhibited a complete or high correlation with the RHE levels (r=1.000, p<0.001), or MCV, MCH, and MCHC in anemia patients (R: 0.575-0.820, p<0.001). ROC curves analyse indicated a highest area under curve of 0.829 (95% CI [0.762-0.895]) and 0.884 (95% CI [0.831-0.936]) for MRV in identifying anemia in male and female patients, respectively (p<0.001). When the optimal cutoff values of MRV were set at 100.95 fl in males and 98.35 fl in females, the sensitivity and specificity were 1.00 and 0.68, and 1.00 and 0.73, respectively. The regression analyse showed that, when being as a categorical variable, MRV showed an odds ratio of 19.111 (95% CI [6.985-52.288]; p<0.001) for the incidence of CRA. The incidence of overall anemia demonstrated a more significant decrease trend along with the increase of MRV quartiles (p-trend < 0.001). This study revealed that the MRV can be used as a convenient and sensitive index in early diagnosis of cancer-related anemia, and decreased MRV level may be the powerful predictor of overt anemia in cancer patients.

Is hemolysis a predictor of complications in patients implanted with a left ventricular assist device?

Aim: Life-threatening complications such as intra-device thrombus, infection, bleeding, kidney and liver failure are frequently encountered due to the use of left ventricular assist devices (LVADs). We aimed to determine the incidence of hemolysis in patients implanted with LVAD, the mechanical complications detected in patients with hemolysis, and whether hemolysis can be used as a predictor of complications. Material and Methods: In this single-center, retrospective clinical research study, we evaluated 40 patients with HeartWare (HeartWare International, Inc., MA, USA) LVAD implants to assess the incidence of hemolysis and its association with mechanical complications. Hemolysis parameters, including lactate dehydrogenase (LDH), bilirubin, serum-free hemoglobin (sfHg), corrected reticulocyte count (CRC), and haptoglobin, were measured in venous blood samples obtained from patients attending cardiovascular outpatient clinics. Patients were divided into two groups( group 1:without hemolysis,n=32; group 2: with hemolysis,n=8) based on the presence or absence of hemolysis, and demographic data, comorbidities, and laboratory results were compared. The time elapsed after implantation of the patients was min: 90 days, max: 3147 days. Results: Hemolysis was detected in 20% (8 out of 40) of the patients included in the study. Patients with hemolysis demonstrated higher CRC, LDH, and total bilirubin levels(p&lt;0,005), and lower haptoglobin levels compared to those without hemolysis(p&lt;0,001). Intra-device thrombus was detected as a mechanical complication in 2 patients with hemolysis. One patient died and the other was discharged after medical treatment. Conclusion: Our findings suggest that routine monitoring of hemolysis parameters in LVAD recipients may aid in the early detection of mechanical complications, such as pump thrombosis, before symptomatic manifestation, thus potentially reducing mortality. Assessing multiple parameters, including LDH, bilirubin, sfHg, CRC, and haptoglobin, is crucial for accurate hemolysis diagnosis and its association with mechanical complications. It should be supported by large series of studies with longer and similar follow-up periods and a larger number of patients.

Eculizumab As a Treatment for Gemcitabine Induced Thrombotic Microangiopathy: A Case Report of Partial Response in a Patient with Pancreatic Adenocarcinoma

Background: Thrombotic microangiopathies (TMAs) involve microvascular platelet thrombi occlusion, often manifesting in dangerous high mortality outcomes. Gemcitabine induced thrombotic microangiopathy, a type of drug induced microangiopathy (DITMA), is an exceedingly rare presentation, currently with no established treatment protocol. The fundamental approach to treatment involves cessation of gemcitabine along with supportive measures. The presentation and pathophysiology of DITMA can be similar to not only Thrombotic Thrombocytopenia Purpura (TTP) but also atypical Hemolytic Uremic Syndrome (aHUS). Therefore, plasmapheresis or anticomplement therapy with eculizumab have been utilized in clinical practice to address potential ADAMTS13 deficiency or complement mediated processes respectively. Case Presentation: A 74-year-old woman with a history of pancreatic adenocarcinoma on gemcitabine (cycle 5 of 6) and autoimmune hepatitis presented from her rehabilitation facility for lethargy, bilateral upper and lower extremity swelling and low platelets. Her labs on presentation were hemoglobin 7.5 g/dL, platelets 39 k/mm3, lactate dehydrogenase (LDH) 1281 U/L, corrected reticulocyte count 2.6% and creatinine 1.53 mg/dL. The peripheral smear was notable for schistocytes. A diagnosis of gemcitabine induced TMA was highest on the differential. The decision was made to initiate weekly eculizumab immediately. Within two days of administrating the first dose of eculizumab the platelet count increased to 67 k/mm3. In terms of further work up, the direct antiglobulin testing was negative and the ADAMTS13 antibody was within normal limits. The LDH, which was above normal in this patient at baseline due to her autoimmune hepatitis, remained elevated throughout her hospital stay. While her platelets and hemoglobin responded appropriately to eculizumab, her renal function continued to deteriorate. She required an ICU stay due to respiratory distress in the setting of volume overload, which necessitated dialysis. Eventually, she developed septic shock. She ultimately passed away on day 18 of her hospital stay, having received 3 total doses of eculizumab. Conclusion: Gemcitabine induced TMA is a rare presentation with no overall standard approach, aside from chemotherapy cessation and supportive measures. Literature on this topic has been limited to case reports. Our case is unique in the approach we utilized based on some of the existing literature. While our treatment approach of weekly eculizumab in suspected gemcitabine induced TMA has been done before, to our knowledge this has typically been done after attempted plasmapheresis. On the other hand, our approach involved immediately starting eculizumab without plasmapheresis due to high suspicion of gemcitabine induced TMA, with TTP eventually being ruled out by the normal ADAMTS13 antibody. We also appreciated an immediate response to the treatment with increased platelets that remained stable, while the renal function did not respond to the treatment. Overall, this case adds to the discussion regarding whether eculizumab can be a promising treatment option for patients with this rare presentation of drug induced TMA in patients with active malignancy undergoing chemotherapy.

Serum Hepcidin Is an Early Predictor of Iron Depletion in Non-Anemic Blood Donors

Introduction Iron deficiency is one of the major concerns in repeated whole blood donors. Although taking iron supplementation, frequent whole blood donors sometimes suffer from iron depletion which contributes to adverse impact on donor's health and may reduce the donor blood pool. Laboratory parameters identifying iron depleted status could help early recognize donors at risk of iron deficiency. Hepcidin is a key iron regulator, which blocks ferroportin to inhibit iron absorption and iron release from hepatocytes. In iron depletion, hepcidin down regulates to encourage iron absorption. Previous data reported that serum hepcidin at less than 10 ng/mL was significantly associated with iron deficiency. According to the biological function and physiological response of hepcidin, we hypothesize that hepcidin could be an early predictor of iron deficiency status in non-anemic individuals. Methods This cross-sectional study was conducted at the Blood Bank Center at Chiang Mai University Hospital, Chiang Mai, Thailand in May 2023. After the informed consent was obtained, ninety-seven healthy repeated whole blood donors who had donated blood at least twice in the past year were recruited. The medical records were reviewed. Basic characteristics and laboratories including complete blood count, reticulocyte indices, iron parameters, and serum hepcidin were evaluated. Donors who had serum ferritin level less than 30 and 30-100 ng/mL were categorized as iron deficiency, and low ferritin group, respectively. Those who had ferritin more than 100 ng/mL were the control group. Logistic regression analysis was done to determine the value of serum hepcidin in predicting iron deficiency status in each group. Results The total of 97 repeated whole blood donors were recruited, 48 (49.5%) were women. Mean age was 34.6 ± 10.7 years. There were 8 donors in the iron deficiency group, 38 in the low ferritin group, and 51 donors in the control group. Mean hemoglobin levels were 13.4 ± 1.1, 13.8 ± 1.2, and 14.2 ± 1.2 g/dL in the iron deficiency, low ferritin group and control, respectively, (p = 0.796). White blood cell and platelet count were not different in these groups. Similarly, red cell indices including mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), and reticulocyte indices showed no difference among groups. Serum hepcidin had a significant difference between the three groups. Median levels of hepcidin were 2.61 (1.37, 3.4) and 12.35 (10.12, 15.57) ng/mL in the iron deficiency and the low ferritin group, respectively, compared to 26.01 (21.36, 39.08) ng/mL in the control group, (p &amp;lt; 0.001). Low serum hepcidin (less than 10 ng/mL) were found in 14 (36.8 %) donors who had ferritin levels less than 100 ng/mL, compared to only 1 (2.4%) donor in the control group, (p &amp;lt; 0.001). In addition, serum hepcidin showed a strong correlation with ferritin levels (r = 0.779). Logistic regression model showed that only female sex (OR: 2.87; 95% CI: 1.26, 6.56) and hepcidin (OR: 0.76; 95% CI: 0.66, 0.86) were significant predictors of repeated donors who were at risk of iron depletion and had the ferritin levels less than 100 ng/mL. Conclusions Frequent blood donors are at risk of iron depletion. Serum hepcidin was a significant predictor of iron depleted status in non-anemic donors. Serum hepcidin also showed a significant correlation with ferritin levels. The downregulation of serum hepcidin was detected prior to the changes of red blood cell indices and reticulocyte responses. Early recognition and identification of donors at risk of iron depletion and prompt iron restoration are the key to ensuring the donor safety and preserving adequate blood donor pool.

Vitamin B12 Deficiency and Hemolytic Anemia Presenting As Pseudo Thrombotic Microangiopathy: A Systematic Review

Introduction: Vitamin B12 deficiency is a common nutritional issue associated with mild anemia and neurological deficits. While rare, hemolytic anemia and low platelet count can be complications of this deficiency, often misdiagnosed as thrombotic microangiopathy (TMA), specifically thrombotic thrombocytopenic purpura (TTP). Pseudo TMA features can occur with B12 deficiency and can be effectively treated with vitamin B12 supplementation. In contrast, true TMA requires plasmapheresis. This systematic review aims to differentiate between pseudo TMA and TMA cases through an analysis of published case reports, providing valuable insights to avoid unnecessary treatments. Methods: We performed systematic review from Pubmed, Web of Science, and Scopus database. Twin independent reviewers (SY and BL) performed the screening using Covidence. Search keywords included “((hemolytic anemia) AND (vitamin B12 deficiency)) OR ((cobalamin deficiency) AND (hemolytic anemia)) Filters: English” for Pubmed, “(((ALL=(Cobalamin deficiency)) OR ALL=(vitamin B12 Deficiency))) AND ALL=(hemolytic anemia) Filter: English” for Web of Science and “( vitamin AND b12 AND deficiency OR cobalamin AND deficiency ) AND hemolytic AND anemia AND ( LIMIT-TO ( LANGUAGE , ”English“ ))” from Scopus. Only case reports in English with full-text availability and adult subjects were selected. Cases without final clinical outcome after treatment were excluded. After a comprehensive review, we identified 43 case reports and series with a total 53 patients that fulfilled our criteria. The details of selection process is illustrated in the prisma diagram (Figure 1). Results: The majority of cases involved males with low hemoglobin and platelet counts upon admission. Evidence of hemolysis was observed, indicated by increased lactate dehydrogenase (LDH), undetectable haptoglobin, and elevated or normal total and direct bilirubin levels. Peripheral blood films confirmed the presence of hemolytic anemia. Due to the co-occurrence of thrombocytopenia and hemolytic anemia, some cases were initially diagnosed as TTP, leading to the initiation of plasmapheresis. In 16 out of 53 reported case reports/series, patient received plasmapheresis. However, low reticulocyte index and normal ADAMTS 13 suggested an alternative diagnosis. Once ADAMTS 13 levels were normal, plasmapheresis was discontinued as TTP was ruled out. Diagnosis of B12 deficiency was confirmed by low B12 levels and elevated methylmalonic acid (MMA) levels. Leading cause for B12 deficiency were pernicious anemia in 28/53 cases, followed by dietary deficiency, and malabsorption due to surgery. Some patients also exhibiting concurrent iron deficiency anemia and folate deficiency. Treatment with subcutaneous B12 injections showed improvement in hemolysis as early as one week. Conclusion: Many cases of Vitamin B12 deficiency presenting with hemolytic anemia and thrombocytopenia are mistakenly diagnosed as TTP, leading to unnecessary and expensive plasmapheresis. Routine B12 level checks should be performed in all patients with hemolytic anemia, as B12 deficiency can be rapidly diagnosed and effectively treated, avoiding misdiagnoses and unnecessary interventions associated with TTP. The short turnover time for B12 levels (hours) compared to ADAMTS 13 activity (1-2 days) further supports the importance of routine B12 assessment in these patients.

A Comparison of Haematological and Biochemical Profiles between Intrauterine Growth Restriction and Normal Piglets at 72 Hours Postpartum.

Intrauterine growth restriction in piglets has been a problem in the pig industry due to genetic selection based on hyperprolificacy. This has led to an increase in the number of underweight piglets and a worsening of the survival rate. The goal of this study was to enhance the knowledge of differences between normal and IUGR piglets a few hours after birth in terms of haematological variables, biochemical parameters, and immunoglobulin levels. Two groups of 20 piglets each were assessed. The control group (N) was made up of piglets with weights greater than 1500 g, and the IUGR group consisted of piglets weighing 500-1000 g and with at least two IUGR features. Blood samples were collected 72 h after birth for analysis of the red and white blood cell parameters, reticulocyte indices, platelet indices, biochemical parameters, and immunoglobulin levels. Alterations in red blood cells and reticulocytes, a lower lymphocyte count, hyperinsulinemia, and high oxidative stress were observed in IUGR piglets (p < 0.05). In contrast, differences were not observed (p > 0.05) in the serum immunoglobulin level. It can be concluded that the haematological and biochemical differences in IUGR piglets with respect to normal-weight piglets are present at birth indicating possible alterations in immunity, metabolism, and redox status; therefore, IUGR piglets could be more vulnerable to illness and future disorders, such as metabolic syndrome.

Abstract 13078: Prognostic Significance of Reticulocyte Production Ability in Patients With Chronic Heart Failure

Background: Anemia is frequently observed in patients with chronic heart failure (CHF), and many studies have evaluated the association between anemia and mortality in CHF. The quality of erythropoiesis is an intrinsic aspect of pathophysiology of anemia, and the ability of reticulocyte production would be one of the pivotal factors in the quality of erythropoiesis. However, it remains unclear whether reticulocyte production ability is associated with clinical outcomes in patients with CHF. Methods: We prospectively examined 1,274 symptomatic CHF patients (mean age 71.9±13.4 years, median LVEF 50 [IQR 36-63]%) in a multicenter registry (25 sites). The reticulocyte production ability was evaluated by calculating the reticulocyte production index (RPI) using reticulocyte counts and serum hematocrit. Patients were divided into four groups according to the presence of WHO defined anemia and the median value of RPI. The primary outcome was composite of all-cause death and hospitalization for worsening HF. Results: During a median follow-up period of 427 (IQR 273-642) days, the primary outcome occurred in 222 (17%) patients. The group with anemia and high RPI (≥0.982, the median) showed the highest incidence of the primary outcome among the groups ( P &lt; 0.001) (Figure A). In an anemic subgroup (n = 611), higher RPI was independently associated with increased adverse events (HR 1.67, 95% CI 1.31-2.13) adjusted for age, sex, estimated glomerular filtration rate, NT-pro BNP, hemoglobin, serum sodium and iron parameters. In this subgroup, erythrocyte counts were not significantly increased in patients with higher RPI compared to lower RPI (Figure B). Moreover, there were no significant differences on transferrin saturation and serum iron between the groups (Figure B). Conclusions: Higher reticulocyte production ability, which might reflect the impaired process of erythropoiesis, was associated with worse clinical outcomes in patients with CHF, especially with anemia.

'Ironing out the risk': Assessing the effect of plateletpheresis donation frequency on iron stores in South-Asian male donors.

Repeated blood donation is a well-known cause of iron deficiency among donors. However, present scientific literature lacks comprehensive evidence regarding the impact of regular plateletpheresis procedures on body iron reserves. In this study, we aimed to detect and correlate iron deficiency (using iron indices) with the frequency of platelet donations. Additionally, we also analysed the correlation between other iron and haematological indices with serum ferritin to determine cost-effective parameters that may serve as an initial screening approach to determine which donors should be subjected to serum ferritin testing. A total of 180 male participants from our platelet donor registry were enrolled in this observational cross-sectional study. Enrolment questionnaires were administered to eligible donors, and biological samples were collected during plateletpheresis donation. Biological tests such as complete blood count, reticulocyte indices, iron indices, vitamin B12 and folate were performed. Donors with ≥12 donations per year showed the highest prevalence of low ferritin (serum ferritin: 15-30 ng/mL) and absent iron stores (serum ferritin <15 ng/mL) (41.3% and 26.7%, respectively). Ferritin showed a significant negative correlation with recent (r = -0.346) and lifetime donations (r = -0.196). The efficacy of other indices for identifying iron depletion was much better using a serum ferritin value <15 ng/mL. Regular plateletpheresis donations can lead to varying severities of non-anaemic iron deficiency. Blood centres must regularly monitor frequent plateletpheresis donors (especially donors with more than 11 donations in a calendar year) and ideally maintain their serum ferritin above 30 ng/mL.

Regenerative response in dogs naturally and experimentally infected with Babesia rossi.

The regenerative response following Babesia rossi infection in dogs is mild, despite severe hemolytic anemia. We aimed to compare the admission absolute reticulocyte count (ARC) and reticulocyte indices in 103 dogs naturally infected with B. rossi with 10 dogs suffering from immune-mediated hemolytic anemia (IMHA) and 14 healthy control dogs. The regenerative response was also evaluated in five dogs experimentally infected with B. rossi. This is a retrospective observational study of records generated on the ADVIA 2120 hematology analyzer. The median hematocrits (HCT) of the B. rossi and IMHA groups were significantly lower than the control group (p < .001 for both); however, no differences were seen between the B. rossi and IMHA groups. Compared with the control group, the median ARC was significantly higher in the B. rossi (p = .006) and IMHA (p = .019) groups but significantly lower in the B. rossi group than the IMHA group (p = .041). In the experimentally infected dogs, there was a sudden decrease in the ARC approximately 48 h after the detection of peripheral parasitemia, which was followed by an increase after treatment. Reticulocytes of naturally infected B. rossi dogs were larger, with more variation in cellular volume. The reticulocytes of the experimentally infected dogs decreased in size with decreasing hemoglobin concentrations as the study progressed. The regenerative response in dogs naturally infected with B. rossi is inadequate, given the severity of the anemia observed, and it might be a result of direct suppressive action by the parasite or host response on the bone marrow.

Reference intervals for reticulocyte indices, immature reticulocyte fraction, and the percentage of hypochromic red blood cells in adult large breed dogs using the ADVIA 2120 hematology analyzer.

Reticulocyte indices are used to characterize anemia, including the identification of regeneration. In people, the immature reticulocyte fraction (IRF), percentage of hypochromic red blood cells (%HYPO-RBC), and other reticulocyte indices have been used as earlier indicators of erythropoiesis and as valuable monitoring tools in the assessment of various therapies. The reference intervals (RI) of the IRF and %HYPO-RBC have not been reported in dogs. The objective of this study was to establish RIs for novel variables (IRF, %HYPO-RBC, and CH-delta) and assess RIs for more commonly reported reticulocyte indices in healthy dogs. RIs were calculated from blood results retrospectively collected from 106 client-owned healthy dogs at the time of induction into a blood donor program using the ADVIA 2120 hematology analyzer (Siemens Healthcare Diagnostics). For the calculation of RIs, appropriate tests were applied for outlier detection and normality assessment. For variables normally distributed, RIs and their respective 90% confidence intervals (CIs) were calculated using parametric methods, while for variables not normally distributed, robust methods were used and bootstrapping for calculating the 90% CIs. The following RIs were established: reticulocyte hemoglobin content (CHr) 24.5-28 pg, mean reticulocyte volume (MCVr) 85.9-99.3 fL, mean corpuscular hemoglobin concentration of reticulocytes (CHCMr) 271.0-306.3 g/L, IRF 10.4%-43.5%, CH-delta 0.5-4.3 pg, and percentage of hypochromic red blood cells (%HYPO-RBC) 0.10%-0.80%. The results of this study provide RIs for novel reticulocyte variables. Further studies are required to determine the clinical utility of IRF, %HYPO-RBC, and CH delta as early indicators of erythropoietic activity in canine patients.

Cold agglutinin syndrome in a patient with human immunodeficiency virus infection

Abstract A 44-year-old female with no prior comorbidities presented with a history of easy fatiguability and yellowish discolouration of eyes of 3 months duration. On examination, she had pallor and icterus. Jugular venous pressure was elevated. Earlier, the patient was diagnosed to have autoimmune haemolytic anaemia and was started on steroids 2 weeks ago elsewhere. Laboratory investigations showed severe anaemia, elevated mean corpuscular volume and lactate dehydrogenase levels. Corrected reticulocyte count was 2.5%. Unconjugated hyperbilirubinaemia was evident. Coomb’s test was positive. In view of suspicion for cold agglutinin syndrome (CAS), monospecific agglutinin test with cold agglutinin titres was done, which tested positive. Serological testing was positive for human immunodeficiency virus (HIV) 1. The patient was diagnosed to have CAS secondary to HIV infection.