Hematology Laboratory Research Articles

Venous blood parameters in determination of respiratory impairment in amyotrophic lateral sclerosis

This study aimed to investigate the relationship between venous blood parameters and respiratory functions in patients with amyotrophic lateral sclerosis (ALS) and develop a model to predict respiratory impairment for individual patients with ALS. A total of 416 ALS patients were included in the study, and various hematologic and biochemical laboratory parameters as well as demographic and clinical factors were collected and compared. A multivariable logistic regression model was constructed to assess the association between FVC and venous blood biomarkers and clinical factors. The results showed that along with onset age, bulbar-onset, disease duration, BMI, eosinophil count (EO#), basophil count (BASO#), creatinine (CREA), uric acid (URCI) and low-density lipoprotein cholesterol/high-density lipoprotein cholesterol (LDL/HDL) ratio were associated with reduced FVC. The area under the ROC curve is 0.735 for the test set and 0.721 for the validation set. The study also developed a relatively acceptable model for predicting respiratory impairment in ALS patients. These findings suggest that EO#, BASO#, CREA, URIC and LDL/HDL ratio can be useful in assessing FVC in ALS and can be easily accessible, accurate, and low-cost parameters.

Pediatric Rattlesnake Envenomations Treated With Crotalidae Equine Immune F(Ab') 2 Antivenom : A 3-Year Retrospective Observational Analysis.

Rattlesnake envenomations are uncommon, and the majority occur in adults. Although Crotalidae equine immune F(ab') 2 antivenom (F(ab') 2 AV; trade name ANAVIP) was introduced in 2018, no pediatric specific studies of F(ab') 2 AV have been reported to date. The objective of this study was to evaluate the clinical performance and adverse effects of F(ab') 2 AV in children. A single-center, retrospective chart review was performed on patients with rattlesnake envenomation presenting to a children's hospital between October 2018 and August 2022. Inclusion criteria were age younger than 18 years and F(ab') 2 AV use. Exclusion criteria were other antivenom use at any time and presentation beyond 24 hours postenvenomation.Demographic characteristics, hemoglobin, platelet count, fibrinogen, international normalized ratio, number of F(ab') 2 AV vials used, infusion-related complications, and clinical outcomes were collected. Twenty-six patients, 19 males and 7 females, with a mean age of 7.7 years (0.67 to 16 years) met inclusion criteria. Fourteen (54%) were treated with only the initial 10 vial F(ab') 2 AV doses. Twelve patients were given additional doses with a median additional vials of 10 (4-34 vials; interquartile range, 8.75-12 vials). The median total vials given for all patients was 10 (10-44 vials; interquartile range, 10-20 vials).Two patients developed acute infusion reactions. Both were treated by slowing the infusion rate and with medications (diphenhydramine, corticosteroids). No delayed reactions were noted. No patients required blood products or surgical interventions.After discharge, no complications, recurrent symptoms, return visits, or readmissions were reported. Follow-up by chart review or phone was obtained for 18 patients, and no postdischarge complications were noted. Seven patients had postdischarge hematologic laboratory evaluations and all were normal. Although limited by small sample size and postdischarge follow-up, F(ab') 2 AV was well tolerated in our series of pediatric patients, consistent with prior studies of all age groups.

Whole blood no-lyse no-wash micromethod for the quantitative measurement of monocyte HLA-DR.

Monocyte (m)HLA-DR expression appears to be a potent marker of immunosuppression in critically ill patients. The persistence of low mHLA-DR expression is associated with an increased risk of nosocomial infections and mortality. To adapt this measurement to pediatric requirements and provide extensive 24/7 access, we have developed a whole blood no-lyse no-wash micromethod (MM) and compared it with the standardized method (SM). mHLA-DR was quantified by flow cytometry using Quantibrite™ Anti-HLA-DR PE/Monocyte PerCP-Cy™5.5 with either the SM performed in a diagnostic hematology laboratory using manufacturer protocol, or a whole blood no-lyse no-wash MM using an Attune flow cytometer located in the pediatric ICU. Median fluorescence intensity was measured in both techniques and converted to antibodies per cell (AB/C) calibrated with BD Quantibrite™ PE beads. Blood and Quantibrite™ reagent volume used with the MM was reduced by 5-fold compared to SM. In addition to Quantibrite™ Anti-Human HLA-DR PE/Monocyte PerCP-Cy™5.5, MM required anti-CD45 and anti-CD19 labeling. We determined the expression of mHLA-DR in 34 patients, 20 adults, and 14 children admitted to ICU. Correlation between MM and SM was excellent (Pearson's correlation: y = 0.8192x + 678.7, r = 0.9270, p < 0.0001). The estimated bias was 2467 ± 1.96 × 3307 AB/C; CI 95% [-4016; +8949]. The no-lyse no-wash whole blood microvolume method for measuring mHLA-DR expression allows for simplified sample preparation without compromising accuracy of the data. This method may simplify immune monitoring of critically ill patient by the deployment of a point of care method.

42008 Long Term Hematologic Laboratory Safety of Dupilumab in Patients Aged 6 Months to 5 Years With Moderate-to-Severe Atopic Dermatitis

42008 Long Term Hematologic Laboratory Safety of Dupilumab in Patients Aged 6 Months to 5 Years With Moderate-to-Severe Atopic Dermatitis

International Society for Laboratory Hematology 2023 Education Issue.

International Society for Laboratory Hematology 2023 Education Issue.

Differences In HbA1c Levels In Diabetes Mellitus Patients Using Capillary And Venous blood Samples

Introduction: HbA1c is a non-enzymatic binding of glucose molecules to hemoglobin through a post- translational glycation process, measuring the level or percentage of glucose bound to hemoglobin, this parameter is used as the main benchmark for controlling Diabetes Mellitus because HbA1c can describe blood sugar levels within a period of 1- 3 months due to red blood cells bound by glucose molecules. HbA1c examination is an examination of glycemic levels which is useful to determine long-term glycemic control in patients with Diabetes Mellitus. From this study to determine differences in HbA1c levels in diabetes mellitus patients using capillary and venous blood samples. Method: Cross-sectional, this research was conducted at the Hematology Laboratory of the Palembang Muhammadiyah Institute of Health and Technology. The population taken was Diabetes Mellitus Patients in the Muhammadiyah Palembang Hospital laboratory. The sample used in this study was 29 samples using purposive sampling. Results: The average HbA1c level was found in 7.91% capillary blood and 7.91% venous blood. Discussion: The conclusion in this study is that capillary and venous blood sampling can be used to check HbA1c levels.

Artificial intelligence and the blood film: Performance of the MC-80 digital morphology analyzer in samples with neoplastic and reactive cell types.

Implementing artificial intelligence-based instruments in hematology laboratories requires evidence of efficiency in classifying pathological cells. In two-Universities, we assessed the performance of the Mindray® MC-80 for hematology patients with frequent leukemic and dysplastic cells. The Mindray MC-80® locates and pre-classifies cells in blood films. In a two-university study, four films were prepared from 591 samples, two each for the analyser MC-80 and the microscope reference method, using reagents from two different manufacturers. We used Microsoft Excel® statistics for imprecision and distributional inaccuracy and a matrix table model (H20-A2 CLSI standard) for sensitivity, specificity and predictive value for atypical cells. The results indicate minimal within-run imprecision (ICSH method) and good intra-method consistency even on duplicate analysis of 413 samples with a high incidence of hematological abnormalities (r = 0.942 or more, except for basophils, r = 0.841, and reactive lymphocytes, r = 0.847). Distributional inaccuracy was also very low compared to the microscope reference, with a pass rate higher than 80% for pathological cells (except 75.1% for reactive lymphocytes). The primary causes of discrepancy were bizarre shapes of dysplastic neutrophils and inconsistent nomenclature for lymphoma cells. Sensitivity for critical samples containing cells typically absent in circulating blood (immature or malignant) was 98.8% for immature granulocytes, 83.8% for all types of neoplastic cells, 93.6% for reactive lymphocytes and 97.5% for nucleated red blood cells. The negative predictive values of MC-80 were 98.8% for immature granulocytes, 88.4% for the different types of neoplastic cells, 97.8% for reactive lymphocytes, and 96.9% for nucleated red blood cells. Our study highlights the outstanding diagnostic performance of this artificial intelligence-based blood film analyzer for hematology patients with circulating abnormal cells. We appreciated the morphological harmonization of cells observed on the screen and those seen in the microscope.

Significant Variability in the Identification and Reporting of Band Neutrophils by Participants Enrolled in the College of American Pathologists Proficiency Testing Program: Time for a Change.

Increased band neutrophils in blood smear differential counts ("bandemia") are entrenched in medicine as a flag for sepsis. However, laboratory hematology experts have long advocated for discontinuation of reporting bands separately from segmented neutrophils because of poor sensitivity and specificity, poor interobserver agreement, and availability of alternative biomarkers for sepsis. To describe band neutrophil reporting practices and reproducibility of band classification among laboratories participating in the College of American Pathologists (CAP) proficiency testing (PT) program. A survey questionnaire was distributed to hematology PT participants. A subsequent morphologic challenge included 12 preselected cell identifications of segmented neutrophils, bands, and metamyelocytes, and a 100-cell manual differential count of a digitally scanned blood smear. Among laboratories that reported manual differentials, most respondents reported bands (4554 of 5268; 86.4%). Only 3222 of 4412 respondents (73.0%) provided band reference ranges. Though participants classified "easy" band neutrophils well (78.0%-98.3%), categorization of cell identifications for "moderate" and "difficult" bands was poor (3.1%-39.0% of laboratories), with classification instead as segmented neutrophils. This pattern was seen regardless of laboratory demographic characteristics. Marked variability in band counts was observed on the 100-cell differential count for both CAP PT participants and CAP Hematology and Clinical Microscopy Committee (HCMC) members (coefficients of variation, 55.8% and 32.9%, respectively). Variability was significantly improved when segmented and band neutrophils were grouped together (coefficients of variation, 6.2% and 5.0%, respectively). Most CAP PT-participating laboratories report band counts, many without reference ranges. The survey confirms significant interlaboratory variability of band enumeration when bands are separately identified from segmented neutrophils. This study reaffirms the CAP Hematology and Clinical Microscopy Committee's strong recommendation to group segmented and band neutrophils together in manual differential counts.

Phase I study evaluating the Fc-optimized FLT3 antibody FLYSYN in AML patients with measurable residual disease

BackgroundAbout half of AML patients achieving complete remission (CR) display measurable residual disease (MRD) and eventually relapse. FLYSYN is an Fc-optimized antibody for eradication of MRD directed to FLT3/CD135, which is abundantly expressed on AML cells.MethodsThis first-in-human, open-label, single-arm, multicenter trial included AML patients in CR with persisting or increasing MRD and evaluated safety/tolerability, pharmacokinetics and preliminary efficacy of FLYSYN at different dose levels administered intravenously (cohort 1–5: single dose of 0.5 mg/m2, 1.5 mg/m2, 5 mg/m2, 15 mg/m2, 45 mg/m2; cohort 6: 15 mg/m2 on day 1, 15 and 29). Three patients were treated per cohort except for cohorts 4 and 6, which were expanded to nine and ten patients, respectively. Primary objective was safety, and secondary efficacy objective was ≥ 1 log MRD reduction or negativity in bone marrow.ResultsOverall, 31 patients were treated, of whom seven patients (22.6%) experienced a transient decrease in neutrophil count (two grade 3, others ≤ grade 2). No infusion-related reaction or dose-limiting toxicity was observed. Adverse events (AEs) were mostly mild to moderate, with the most frequent AEs being hematologic events and laboratory abnormalities. Response per predefined criteria was documented in 35% of patients, and two patients maintained MRD negativity until end of study. Application of 45 mg/m2 FLYSYN as single or cumulative dose achieved objective responses in 46% of patients, whereas 28% responded at lower doses.ConclusionsFLYSYN monotherapy is safe and well-tolerated in AML patients with MRD. Early efficacy data are promising and warrant further evaluation in an up-coming phase II trial.Trial registration This clinical is registered on clinicaltrials.gov (NCT02789254).

Conversion between sirolimus and everolimus in heart transplant recipients.

Sirolimus and everolimus are mechanistic target of rapamycin inhibitors (mTORi) that may be included in immunosuppression regimens for orthotopic heart transplant (OHT) recipients. mTORi play a role in slowing progression of cardiac allograft vasculopathy; however, they have poor tolerability, sometimes necessitating a change between agents or therapies. The literature surrounding a conversion between mTORi are incongruent, thus this study was designed to assess the concentration/dose ratio for each medication around the time of conversion to provide guidance for a conversion strategy between mTORi. We conducted a retrospective study of adult OHT recipients who were maintained on both sirolimus- and everolimus-based immunosuppression regimens. The primary outcome was the concentration/dose (C/d) ratio of sirolimus to everolimus. Secondary outcomes included changes in hematologic and lipid labs and patient-reported intolerances. The C/d ratio of sirolimus was 4.42, whereas the everolimus ratio was 2.23, resulting in a sirolimus: everolimus ratio of 1.98. Secondarily, after converting between mTORi, 93% of patients who reported intolerance(s) to one agent had a resolution of that intolerance. In our patient population, everolimus appeared better tolerated than sirolimus, with significantly more patients having no reported intolerances and significantly fewer patients experiencing edema. Other lab values were similar between patients receiving sirolimus and everolimus, except for an increased hemoglobin level in those receiving everolimus. The conversion ratio of 1:2 observed in our population suggests OHT recipients may require an increased dose of EVL compared with SRL to maintain the same goal trough levels. A conversion between mTORi appeared to improve tolerability and did not lead to clinically significant worsening of any measured lab value.

A Comparative Evaluation of Performance of Sysmex XN 3000 and Horiba Yumizen H2500 Automated Complete Blood Count Analysers

Modern automated laboratory haematology analysers use various methods to measure different haematological parameters. These parameters are useful in the diagnostic and clinical interpretation of patient symptoms. So, it is very important to compare the performance of different analysers measuring the same parameter. Hence, a comparison of complete blood counts analysed by Sysmex XN 3000 and Horiba Yumizen H2500 was performed. Total 296 EDTA anti-coagulated blood samples were processed in both the analysers in duplicate within 4 h of collection. The white blood cell count, red blood cell count, erythrocyte indices, differential leukocyte count, platelet count and platelet indices and reticulocyte count were compared. A good level of correlation and agreement between different parameters were obtained. A strong correlation was observed (r > 0.9) between Sysmex XN 3000 and Yumizen H2500 for WBC (0.997), RBC (0.997), Haemoglobin (0.999), haematocrit (0.974), MCV (0.902), MCH (0.99),, platelet count by impedance (0.989), mean platelet volume (0.954), plateletcrit (0.971), platelet distribution width (PDW) (0.916), neutrophils (0.997), lymphocytes (0.989), monocytes (0.943), and eosinophils (0.991) counts. A moderate correlation was observed for RDW-CV (0.75). The basophils count showed poor correlation (r < 0.5) possibly because of sample selection with mostly low basophils count. An acceptable bias was observed for most of the parameters like WBC, RBC, Haemoglobin, Haematocrit, platelet counts, neutrophils, lymphocytes, eosinophils and monocytes. The studied instruments ensured satisfactory interchangeability except for few parameters, thus facilitate substitution of one analyser by another without affecting the clinical decision making.

Clinical utility of peripheral blood laboratory testing in the diagnostic workup of prurigo nodularis: A multicenter cohort study

Prurigo nodularis (PN) is a chronic inflammatory skin disease associated with several systemic comorbidities. However, there is lack of evidence supporting specific laboratory testing in the diagnostic workup of PN patients. To characterize the frequency and severity of clinical laboratory abnormalities in PN patients compared to controls. Cross-sectional study of adult patients between October, 2015 and August, 2021 using TriNetX, a global health records database encompassing over 74 million patients. A total of 12,157 PN patients were matched to 12,157 controls. Significantly, greater proportions of PN patients had moderate-to-severely decreased hemoglobin, elevated transaminases, decreased albumin, increased bilirubin, increased serum creatinine, decreased estimated glomerular filtration rate, higher hemoglobin A1c levels, and alterations in thyroid stimulating hormone. Our data identifies associated laboratory abnormalities in PN patients but is unable to support a causal relationship. PN patients are more likely to have laboratory abnormalities on renal, hepatic, hematologic, endocrine, and metabolic laboratory testing, demonstrating a role for systemic testing in the diagnostic workup of PN patients.

Interference of red cell parameters by an endogenous substance: An interesting case report

A 6-month-old infant was brought to the hospital with fever, vomiting, loose stools, and abdominal distension. The venous blood sample was sent to the hematology laboratory for complete blood count (CBC). Analysis of the sample on the automated hematology analyzer showed a discrepancy in red blood cell count, hemoglobin, and hematocrit values. Examination of the blood sample revealed a cherry red color appearance. On centrifugation, the plasma was lactescent. This finding provided an essential clue to the patient’s diagnosis of severe hypertriglyceridemia. Turbidity of lactescent plasma interferes with CBC. Analytical errors in the CBC using an automated hematology analyzer caused by lactescent plasma and methods to resolve the false values will be discussed in this case report.

Positive direct antiglobulin tests in cancer patients on immune checkpoint inhibitors: A case series from India.

Tumour cells activate immune checkpoints such as programmed death receptor-1 (PD-1) and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) signalling pathways to inhibit T lymphocyte activation and thus escape from immune surveillance. Immune checkpoint inhibitors (ICPis) reactivate T lymphocytes to recognize cancer cells by blocking CTLA-4 or PD-1. Autoimmune haemolytic anaemia is a rare, but often severe, complication of ICPis. Therefore, we performed a retrospective clinical case review, including serologic, haematology, and biochemistry laboratory results, of three patients who developed autoantibodies to erythrocytes following treatment with pembrolizumab, an anti-PD-1 inhibitor. Serologic testing of blood samples from these patients showed their red cells were positive for direct antiglobulin test (DAT+for IgG in two cases and IgG with C3d in one case). Antibody detection test was negative. No patient had clinical and laboratory features of haemolysis. There were no additional immune-related adverse events. IgG antibodies coating red cells were neither IgG1 nor IgG3 in class and elution was found negative in all. In conclusion, immunohaematology laboratories should be aware of the possibility of erythroid autoantibodies and their nature in cancer patients receiving ICPis. The result of a positive DAT should be interpreted carefully in these patients to exclude other possible causes of anaemia.

Pooled Safety Analysis of Single-Agent Lurbinectedin in Patients With Advanced Solid Tumours

BackgroundLurbinectedin was approved by FDA and other health regulatory agencies for treating adults with metastatic small cell lung cancer (SCLC) with disease progression on or after platinum-based chemotherapy. Safety profile at approved dose (3.2 mg/m2 every 3 weeks) was acceptable and manageable in 105 adult SCLC patients from a phase II basket trial. This study analyses safety data from several solid tumours treated at the lurbinectedin-approved dose. MethodsData were pooled from 554 patients: 335 from all nine tumour-specific cohorts of the phase II basket trial and 219 from a randomised phase III trial (CORAIL) in platinum-resistant ovarian cancer. Events and laboratory abnormalities were graded using NCI-CTCAE v.4. ResultsMost common tumours were ovarian (n = 219, 40%), SCLC (n = 105, 19%) and endometrial (n = 73, 13%). Transient haematological laboratory abnormalities were the most frequent grade 3 or more events: neutropenia (41%), leukopenia (30%), anaemia (17%) and thrombocytopenia (10%). Most common treatment-emergent non-haematological events (any grade) were transient transaminase increases (alanine aminotransferase [66%], aspartate aminotransferase [53%]), fatigue (63%), nausea (57%), constipation (32%), vomiting (30%) and decreased appetite (25%). Dose reductions were mostly due to haematological toxicities, but most patients (79%) remained on full lurbinectedin dose. Serious events mostly consisted of haematological disorders. Eighteen treatment discontinuations (3%) and seven deaths (1%) were due to treatment-related events. ConclusionsThis analysis confirms a manageable safety profile for lurbinectedin in patients with advanced solid tumours. Findings are consistent with those reported in patients with relapsed SCLC, Ewing sarcoma, germline BRCA1/2 metastatic breast cancer, neuroendocrine tumours and ovarian cancer.

Reference Ranges and Gender Differences in Blood Parameter Establishment at Dental Institutions: A Prospective Study

Gender differences in haematological variables were shown to be considerable in the majority of these research; as such, determining this difference and estimating the population’s reference range are crucial. The goal of the current study was to establish a range of comparison for the general population as well as to evaluate the haematological variations in several blood parameters between males and females. MATERIAL AND METHODOLOGY: The research was organised on a total of 250 patients with 125 males (50%) and 125 females (50%) in the haematology laboratory of the institution. 20 µl of capillary blood obtained by puncturing the skin over a finger was introduced into 100 µl of diluent. The contents of diluent are sodium sulfate, sodium chloride, a buffering agent, EDTA, formaldehyde, 1pyridone-2-sulphur a with pH of the 6.2-7.6. RESULTS: The study shows that the mean values of Red blood cells, Haemoglobin, Haematocrit, Mean corpuscular volume and Mean corpuscular Haemoglobin were statistically substantially reduced in men compared to females (p &lt; 0.001), and that the mean values of MCHC were considerably lower in females than in males (). PLT and PCT, two haematological markers, were marginally higher in females than in men, with statistically significant findings at p &lt; 0.05. CONCLUSION: Compared to only reporting cell counts, the CBC provides a wealth of detailed information about every blood cell. Even if there are new molecular analysis-based diagnostic and prognostic techniques available today, it’s still crucial to recognize the significance of the tests that doctors have been prescribing for decades. Our research would contribute to the updating of current data on blood variables, their reference range, and gender variance at the regional level.

Physico-chemical quality of ashes post-incineration and waste from laboratory automaton biochemistry, hematology and serology in Togo

Biochemistry, hematology and serology laboratories produce very heterogeneous discharges capable of polluting water, soil and the environment. The aim of this study is to evaluate the physico-chemical quality of the ashes, effluents and the range of equipment that produce the much effluent and pollutions.This is a descriptive and cross-sectional study of 82 laboratories followed by the sampling of effluents from automatons. The physico-chemical parameters and trace elements are determined according to French Standardization Association (AFNOR) standards and used atomic absorption spectrometers and molecular absorption spectrometers. The Mindray hematology automaton is used for 64.7% and 81.5% of the cans are used to collect waste. Semehow 77.8% of discharges are released into sinks without adequate treatment in 42.6% of cases. Chemical analyses show that trace elements (TE) levels in effluents range from 5.89 ± 0.21 to 105.80 ± 0.96 μg/L for Arsenic; from 5.62 ± 0.34 to 6.53 ± 0.49 μg/L for Cadmium; 0.70 ± 0.01 to 10.63 ± 1.17 μg/L for Mercure and 35.72 ± 1.88 to 61.98 ± 5.15 μg/L for Plomb. pH ranges from 7.17 ± 0.65 to 8.3 ± 1.35 with total dissolved salts (TDS) range from 3260.20 ± 401.68 to 17117.41 ± 265.37 mg/L. All averages of contamination indices (CI) and polymetallic contamination indices (PCI) for post-incineration ashes are greater than 1.5 with the exception of Hg (0.22). For effluents, in addition to polymetallic contamination indices (PCI), CI in hematology effluents (1.09), all CI are less than 1. Hematology effluents have a lower pH and are much more mineralized and concentrated in trace elements while those from biochemistry and serology appear to be similar. However, the effluents of Biochemistry and Serology are more basic and concentrated in Suspended Solids with higher values obtained in Biochemistry. Although the physico-chemical parameters comply with WHO standards, discharges from automatons deserve appropriate treatment before their disposal.

High-throughput microfluidic blood testing to phenotype genetically linked platelet disorders: an aid to diagnosis

AbstractLinking the genetic background of patients with bleeding diathesis and altered platelet function remains challenging. We aimed to assess how a multiparameter microspot-based measurement of thrombus formation under flow can help identify patients with a platelet bleeding disorder. For this purpose, we studied 16 patients presenting with bleeding and/or albinism and suspected platelet dysfunction and 15 relatives. Genotyping of patients revealed a novel biallelic pathogenic variant in RASGRP2 (splice site c.240-1G>A), abrogating CalDAG-GEFI expression, compound heterozygosity (c.537del, c.571A>T) in P2RY12, affecting P2Y12 signaling, and heterozygous variants of unknown significance in the P2RY12 and HPS3 genes. Other patients were confirmed to have Hermansky-Pudlak syndrome type 1 or 3. In 5 patients, no genetic variant was found. Platelet functions were assessed via routine laboratory measurements. Blood samples from all subjects and day controls were screened for blood cell counts and microfluidic outcomes on 6 surfaces (48 parameters) in comparison with those of a reference cohort of healthy subjects. Differential analysis of the microfluidic data showed that the key parameters of thrombus formation were compromised in the 16 index patients. Principal component analysis revealed separate clusters of patients vs heterozygous family members and control subjects. Clusters were further segregated based on inclusion of hematologic values and laboratory measurements. Subject ranking indicated an overall impairment in thrombus formation in patients carrying a (likely) pathogenic variant of the genes but not in asymptomatic relatives. Taken together, our results indicate the advantages of testing for multiparametric thrombus formation in this patient population.

Psychological toxicity in classical hematology.

Although considered "benign," mild blood count abnormalities, genetic factors imparting inconsequential thrombotic risk, and low-risk premalignant blood disorders can have significant psychological and financial impact on our patients. Several studies have demonstrated that patients with noncancerous conditions have increased levels of anxiety with distress similar to those with malignancy. Additionally, referral to a classical hematologist can be a daunting process for many patients due to uncertainties surrounding the reason for referral or misconstrued beliefs in a cancer diagnosis ascribed to the pairing of oncology and hematology in medical practice. If not properly triaged, incidental laboratory abnormalities can trigger extensive and costly evaluation. These challenges are compounded by a lack of consensus guidance and generalizability of modern reference ranges that do not adequately account for common influencing factors. Although often benign, incidental hematologic findings can lead to emotional suffering and careful consideration of the potential psychological and financial duress imparted to an individual must be considered. In this article, we will review the current literature describing the psychological effect of some commonly known hematologic conditions, identify benign causes for variations in hematologic laboratory values, and provide recommendations to reduce psychological toxicity as it pertains to hematologic testing.

Whole F8 gene sequencing combined with splicing functional analyses led to a substantial increase of the molecular diagnosis yield for non-severe haemophilia A.

Conventional genetic investigation fails to identify the F8 causal variant in 2.5%-10% of haemophilia A (HA) patients with non-severe phenotypes. In these cases, F8 deep intronic variants could be causal. To identify pathogenic F8 deep intronic variants in genetically unresolved families with non-severe HA analysed in the haematology laboratory of the Hospices Civils de Lyon. The whole F8 was analysed by next generation sequencing. The pathogenic impact of candidate variants identified was assessed using both in silico analysis (MaxEntScan and spliceAI) and functional analysis (RNA or minigene assay). Sequencing was performed in 49/55 families included for which a DNA sample from a male propositus was available. In total, 33 candidate variants from 43 propositi were identified. These variants corresponded to 31 single nucleotide substitutions, one 173-bp deletion, and an 869-bp tandem triplication. No candidate variant was found in six propositi. The most frequent variants found were the association of [c.2113+1154G>C and c.5374-304C>T], identified in five propositi, and the c.2114-6529C>G identified in nine propositi. Four variants had been previously described as HA-causing. Splicing functional assay found a deleterious impact for 11 substitutions (c.671-94G>A, c.788-312A>G, c.2113+1154G>C, c.2114-6529C>G, c.5999-820A>T, c.5999-786C>A, c.5999-669G>T, c.5999-669G>A, c.5999-669G>C, c.6900+4104A>C, and c.6901-2992A>G). The HA-causing variant was identified in 33/49 (67%) cases. In total, F8 deep intronic variants caused 8.8% of the non-severe HA among the 1643 families analysed in our laboratory. The results emphasise the value of whole F8 gene sequencing combined with splicing functional analyses to improve the diagnosis yield for non-severe HA.