Bone marrow aspiration in a patient with systemic microsporidium

A 34-year-old female presented with several weeks of fever, fatigue, weight loss, abdominal pain and hemoptysis. PE revealed moderate pallor, RUQ pain, mild dyspnea, conjunctival injection and hepatomegaly. The CBC showed anemia, mild leukocytosis, hypoalbuminemia, hypertransaminasemia, presence of nucleated red blood cells. Microsporidium was found in BMA.

PERFORMANCE EVALUATION OF NUCLEATED RED BLOOD CELL (NRBC) COUNT USING A FULLY AUTOMATED HAEMATOLOGY ANALYZER VERSUS MANUAL COUNTING

Objective: To evaluate the performance of Nucleated RBC (NRBC) Count using a fully automated haematology analyzer versus manual counting. Study Design: Cross-Sectional Study. Place and Duration of Study: Department of Hematology, Armed Forces Institute of Pathology, from Sep 2019-Jun 2020. Methodology: Routine fresh whole blood samples were run on Sysmex XN-3000 automated haematology analyzer and 384 samples with results of ≥0.1% Nucleated red blood cells were included in this study. Manual NRBC counting was carried out twice on Leishman-stained peripheral blood smears from all 384 samples. Comparison between manual and automated nucleated red blood cell counting methods was statistically analyzed through linear regression analysis & coefficient correlation. The degree of agreement between two methods was analyzed through Bland-Altman plot. Finally, concordance between the two methods was also analyzed at 5 different ranges of nucleated red blood cells. Results: Linear regression analysis revealed a (r2) value of 0.97. Regression equation was calculated as XN = 0.76MC ± 1.28, with 95% limits of agreement between ± 40.42% and -24.47%. A mean bias of 7.97% was demonstrated through Bland-Altman plot. Concordance analysis revealed a concordance rate of 93.74% (360/384). Nucleated red blood cell counting between two methods were more concordant when nucleated red blood cell counts were <200%. Conclusion: Nucleated red blood cells counting by XN-3000 automated hematology analyzer is statistically comparable to manual nucleated red blood cell counting. We suggest that automated counting can be adopted in routine hematology laboratory as a replacement of manual NRBC counting.

An Early Diagnosis of Thalassemia: A Boon to a Healthy Society

The β-thalassemia is a hereditary blood disorders, characterized by reduced or absent synthesis of the hemoglobin beta chain that cause microcytic hypochromic anemia. An early diagnosis, economical test, awareness programs and prenatal screening will be a milestone for the eradication of this genetic disorder and to reduce burden of the health sector of a country subsequently the economics. Initially, the diagnosis of β-thalassemia depends on the hematological tests with red cell indices that disclosed the microcytic hypochromic anemia.Hemoglobin analysis shows the abnormal peripheral blood smear with nucleated red blood cells, and reduced amounts of hemoglobin A (HbA). In severe anemia, the hemoglobin analysis by HPLC reveals decreased quantities of HbA and increased the level of hemoglobin F (HbF).The decrease level of MCV and MCH are also associated with β-thalassemia. There are various different molecular techniques such as ARMS PCR, allele-specific PCR, Gap PCR, denaturing gradient gel electrophoresis, reverse dot blotting, DGGE, SSCP, HRM, MLPA, sequencing technology and microarray available to identify the globin chain gene mutations. These molecular techniques can be clustered for detection by mutation types and alteration in gene sequences.

Nucleated red blood cells as a biomarker for mortality in infants and neonates requiring veno-arterial extracorporeal membrane oxygenation for cardiac disease

Introduction: Nucleated red blood cells (NRBC) are rare in the peripheral circulation of healthy individuals and their presence have been associated with mortality in adults and very low birth weight newborns, however, its value as a biomarker for mortality in infants requiring veno-arterial (VA) extracorporeal membrane oxygenation (ECMO) has yet to be studied. We sought to determine if NRBC can serve as a biomarker for ECMO mortality and inpatient mortality in infants requiring V-A ECMO. Methods: A single-center retrospective chart review analyzing infants <1 year of age requiring VA ECMO due to myocardial dysfunction or post-cardiotomy between January 1, 2011 to June 30, 2020. Results: One hundred two patients required VA ECMO. Sixty-five patients required ECMO post-cardiotomy, 19 for perioperative deterioration, and 18 for myocardial dysfunction. Fifty-one patients (50%) died (21 died on ECMO, 30 died post-ECMO decannulation). Multivariable analysis found Age < 60 days (OR 13.0, 95% CI 1.9–89.6, p = 0.009), NRBC increase by >50% post-ECMO decannulation (OR 17.1, 95% CI 3.1–95.1, p = 0.001), Single Ventricle (OR 9.0, 95% CI 1.7–47.7, p = 0.01), and lactate at ECMO decannulation (OR 3.0, 95% CI 1.3–7.1, p = 0.011) to be independently associated with inpatient mortality. ROC curves evaluating NRBC pre-ECMO decannulation as a biomarker for mortality on ECMO (AUC 0.80, 95% CI 0.68–0.92, p ⩽ 0.001) and post-ECMO decannulation (AUC 0.75, 95% CI 0.65–0.84, p ⩽ 0.001) show NRBC to be an accurate biomarker for mortality. Conclusions: Greater than 50% increase in NRBC post-ECMO decannulation is associated with inpatient mortality. NRBC value pre-ECMO decannulation may be a useful biomarker for mortality while on ECMO and post-decannulation.

Reference Values to Assess Hemodilution and Warn of Potential False-Negative Minimal Residual Disease Results in Myeloma

Background: Whereas, in most patients with multiple myeloma (MM), achieving undetectable MRD anticipates a favorable outcome, some others relapse shortly afterwards. Although one obvious explanation for this inconsistency is the use of nonrepresentative marrow samples due to hemodilution, there is no guidance on how to evaluate this issue. Methods: Since B-cell precursors, mast cells and nucleated red blood cells are normally absent in peripheral blood, we analyzed them in 1404 bone marrow (BM) aspirates obtained in numerous disease settings and in 85 healthy adults (HA). Results: First, we confirmed the systematic detection of the three populations in HA, as well as the nonreduced numbers with aging. Pairwise comparisons between HA and MM patients grouped according to age and treatment showed significant variability, suggesting that hemodilution should be preferably evaluated with references obtained from patients treated with identical regimens. Leveraging the MRD results from 118 patients, we showed that a comparison with HA of similar age could also inform on potential hemodilution. Conclusions: Our study supports the routine assessment of BM cellularity to evaluate hemodilution, since reduced BM-specific cell types as compared to reference values (either treatment-specific or from HA if the former are unavailable) could indicate hemodilution and a false-negative MRD result.

Histopathological Effects and Micronucleus Assay of Glyphosate-Based Herbicides on Cultured African Catfish (Clarias Gariepinus, Burchell 1822)

Background: Glyphosate, a brand of agricultural herbicides which intensive use has led to widespread contamination of different ecosystems. This study was designed to determine both organotoxicity and genotoxicity of glyphosate on African Catfish (Clarias gariepinus) exposed to different concentrations for 96 hours.Methods: Questionnaires were administered (physically and online) to determine the type of glyphosate-based herbicides mostly used by fish farmers. Seventy-five apparently healthy adult Clarias gariepinus (300g) were sourced from a local farmer, transported in a plastic keg to Fish and Wildlife Laboratory at the Department of Veterinary Public Health and Preventive medicine for two (2) weeks of acclimatization. After acclimatization, they were further divided into four (4) groups (T0 (Control), T1 (0.003ml/L), T2 (0.0045ml/L) and T3 (0.006ml/L)) by simple randomization and each group replicated into three (3) treatments. At the expiration of 96 hours of exposure, blood samples and organs (Gills, Kidney, and Liver) were collected for mononuclear assay and histopathological lesions respectively.Results: Exposed groups showed erratic swimming, splashing, and restlessness. Mortalities rate was dose-dependent (two (2) mortalities at 0.0045ml/L concentration (T2) and five (5) mortalities 0.006ml/L concentration (T3)). Observed histopathological lesions occurred at higher dose treatment (0.0045ml/L (T2) and (0.006ml/L) T3)) concentrations. The gills showed diffuse stunted and eroded secondary lamellae and severe congestion of the blood channel at the core of the primary lamellae. Lesions in the Liver include severe diffuse vacuolation of the hepatocytes, moderate to severe portal congestion and mild diffuse vacuolation of hepatocytes and moderate diffuse vacuolation of hepatocytes, and severe portal congestion. In the kidney, there was mild to moderate congestion of the interstitium and focus of interstitial oedema within the parenchyma. There was presence of micronucleus in the fish nucleated red blood cells at higher dose concentrations.Conclusion: This study showed that Glyphosate-based herbicides are highly toxic to Clarias gariepinus, therefore their use near the fish farm or in areas close to the aquatic environment should be discouraged. The agricultural community should also be conscious of the potentially adverse effects of pesticides. This is to prevent the water body from the residue of herbicides that would have washed down to the water.

Comparing Nucleated Red Blood Cells in Neonates with and without Retinopathy

Introduction: Retinopathy of prematurity (ROP) is one of the reasons of blindness in children. Numerous predisposing factors have been reported, including prematurity and oxygen therapy. We assessed the number of nucleated red blood cells in neonates with ROP and compared it with the patients that didn t suffer from the situation. Methodology: This case-control study was performed on 151 neonates with birth weight<1500 gr and/or gestational age< 32 weeks that were born in Ghaem Hospital of Mashhad from 2017 to 2020. A Cell Blood Count (CBC) sample and peripheral blood smear was taken. The Number of nucleated red blood cells or NRBC per 100 white blood cells was reported. Complete characteristics of neonates (birth weight, sex, gestational age, Apgar Score), mother's history (age, pregnancy and labor problems, kind of delivery, and pariety), and lab results were collected and recorded in the checklist. The assessment of the patients were continued until they were discharged and then they got visited by an ophthalmologist at the age of 32 weeks or four weeks after birth in terms of ROP and followed up. Control group included premature neonates without ROP. Results: Among of 151 neonates studied, 47 cases (24.9%) were normal and 104 ones (75.1%) had retinopathy of prematurity. Differences of measured factors in two groups of neonates with and without ROP were as followed: NRBC/100 (P = 0.009), absolute NRBC (P = 0.465), Apgar score of first (P = 0.131) and fifth (P = 0.002) minutes. Conclusion: Increasing umbilical cord NRBC/100 of neonates along with other available methods can be useful as a predictor factor of ROP and it can also predict the ROP intensity.

Nucleated red blood cells as a biomarker for mortality in neonates following cardiac surgery

Introduction: Nucleated red blood cells (NRBCs) are immature red cells that under normal conditions are not present in the peripheral circulation. Several studies have suggested an association between elevated NRBC and poor outcome in critically ill adults and neonates. We sought to determine if elevations in NRBC value following cardiac surgery and following clinical events during the hospital stay can be used as a biomarker to monitor for mortality risk in neonates post-cardiac surgery. Materials and methods: We constructed a retrospective study of 264 neonates who underwent cardiac surgery at Children’s Hospital, New Orleans between 2011 and 2020. Variables included mortality and NRBC value were recorded following cardiac surgery and following peri-operative clinical events. The study was approved by LSU Health IRB. Sensitivity, specificity, receiver operating characteristic (ROC) curves with area under the curve (AUC) and logistic regression analysis were performed. Results: Thirty-six patients (13.6%) died, of which 32 had an NRBC value ≥10/100 white blood cell (WBC) during hospitalisation. Multi-variable analysis found extracorporeal membrane oxygenation use (OR 10, 95% CI 2.9–33, p=<0.001), NRBC ≥10/100 WBC (OR 16.1, CI 4.1–62.5, p ≤ 0.001) and peak NRBC in the 14-day period post-cardiac surgery (continuous variable, OR 1.05, 95% CI 1.0–1.09, p = 0.03), to be independently associated with mortality. Using a cut-off NRBC value of 10/100 WBC, there was an 88.9% sensitivity and a 90.8% specificity, with ROC curve showing an AUC of 0.9 and 0.914 for peak NRBC value in 14 days post-surgery and entire hospitalisation, respectively. Conclusions: NRBC ≥10/100 WBC post-cardiac surgery is strongly associated with mortality. Additionally, NRBC trend appears to show promise as an accurate biomarker for mortality.