BMC Developmental Biology

Abstract Objectives Intraoperative PTH (IOPTH) can be challenging to offer through central laboratories despite its clinical benefit. We describe the implementation of a central laboratory-based IOPTH assay and workflow in a tertiary care centre. Methods The Elecsys® PTH STAT assay was assessed in EDTA plasma on the Cobas® e411 analyzer. Assay validation included precision, linearity, coefficient of variation (CV), accuracy, stability, and dilution. Samples were transported to the central laboratory and resulted via telephone to the operating room. We describe a case series of patients with primary hyperparathyroidism (PHPT) who underwent parathyroid surgery using our described IOPTH workflow. Results Within- and between-day CV was ≤3.0 % for quality control material that ranged from 2.2–44.6 pmol/L. Passing–Bablok regression yielded a slight proportional negative bias between the two Cobas e411 instruments [Elecsys® PTH our centre=0.95 (95 % CI: 0.90–1.00) × Elecsys® PTH Toronto − 0.05 (95 % CI: −0.20 to 0.09) (n=22)], but high correlation (r=0.99) as compared to PTH measured on the Vitros® XT 7600 analyzer [Elecsys® PTH=0.91 (95 % CI: 0.73–1.1) × Vitros® PTH + 0.1 (95 % CI: −0.34 to 0.76), r=0.96 (n=40)]. The mean operating time across ten patients surgically cured for PHPT was 47.1 min (±9.1) and no patients required intraoperative frozen tissue analysis. Conclusions The Elecsys® PTH STAT assay demonstrated acceptable analytical performance, and the described IOPTH workflow was implemented successfully via a collaborative hospital-wide initiative. We discuss our model to help guide other institutions in implementing and improving IOPTH workflows.

Abstract Objectives Myelodysplastic neoplasms and dysplastic chronic myelomonocytic leukemia are characterized by cytopenia. Therefore, transfusion dependency is high in these dysplastic neoplasms. We investigated the impact of molecular genetics on the transfusion dependency in dysplastic neoplasms. Methods We investigated the impact of the myeloid mutation burden on transfusion dependency in myelodysplastic neoplasms and dysplastic chronic myelomonocytic leukemia. In addition, the effect of different functional genetic groups, such as spliceosomes and epigenetic regulator gene mutations, on transfusion dependency was assessed in these patients. Confounding transfusion triggers were ruled out by the patient selection criteria and regression analyses. Results A greater number of mutations lead to a higher transfusion dependency for red blood cells and platelet concentrates. A higher transfusion dependency was associated with a higher transformation to acute myeloid leukemia. Spliceosome mutations were associated with a higher transfusion dependency of red blood cell concentrates than epigenetic regulator mutations. Conclusions Molecular genetics has the potential to improve the precision of patient blood management in dysplastic neoplasms.

Abstract Objectives A variety of methods are currently used to measure von Willebrand factor (VWF) activity, but still the VWF ristocetin cofactor (VWF:RCo) assay using the manual aggregometry technique is the reference method, even having high inter-laboratory variability. The automated coagulation analyzers offer several advantages for routine testing. Herein the performance of the automated Sysmex CS2000/2100i analyzer was compared to the manual aggregometer Chrono-log 700 for assessing VWF:Co activity in patients suspected of having von Willebrand disease (VWD). Methods Plasma samples from 136 patients were prospectively collected, and blindly analyzed on both instruments, simultaneously. Linear regression analysis, Bland-Altman test, intra-class correlation coefficient (ICC), and area under receiver-operator characteristic (ROC) curve were used to evaluate the performance of the automated VWF:RCo assay. Results There was a strong positive correlation between the two assays (r=0.86, p<0.0001) with an excellent reliability ICC value of 0.81 (95 % CI: 0.74–0.86). A very good degree of agreement between the two assays was also evidenced with an estimated bias of −0.055 (−0.58 to 0.46). The ROC curve for the automated VWF:RCo assay was 0.86 (95 % CI: 0.78–0.92; p<0.0001). Using a cut-off value of 0.44 UI/mL for VWF:RCo activity, the sensitivity and specificity values were 91.2 %, and 88.2 % for the automated assay. The positive and negative positive values for VWD detection were 72.9 %, and 96.7 %, respectively. Conclusions Collectively, these findings indicate that the automated VWF:RCo assay yields comparable results to the manual aggregometry assay, with very good accuracy and precision to help diagnose patients suspected with VWD.

Abstract Objectives This study aims to investigate the frequency, detection, and distribution of platelet clumps, assess the performance of automated digital microscopy (Cellavision) for detecting platelet clumps and explore strategies to optimize detection efficiency. Methods We conducted a retrospective analysis of 987,586 hemograms to evaluate the frequency of platelet clumps, and a study of 246 hemograms with platelet clumps for manual and digital microscopic reviews of blood smears. We investigated the locations and sizes of these clumps along the smear, and evaluated the detection capacity of the Cellavision system. Results Platelet clumps were found in 0.29 % of cases, with a higher incidence in pediatric and elderly populations. Platelet clumps were more frequent and larger at the smear periphery. Cellavision achieved 93 % sensitivity when combining the leukocyte and red blood cell observation fields. The strategy of reviewing only selected cases (platelet count <50 × 109/L or history of clumps) detected 97 % of platelet clumps, while reducing manual reviews by fourfold. Conclusions Automated digital microscopy is an effective tool for detecting platelet clumps, but it requires manual review in specific cases. Expanding image acquisition to the feather edge could further improve detection. A combined approach maximizes efficiency and ensures diagnostic accuracy, particularly in critical cases with low platelet counts.

Abstract Objectives To investigate the diagnostic value of serum soluble endorphin (sENG) combined with BISAP score for severe acute pancreatitis (SAP) complicated with septic shock. Methods A total of 150 cases of SAP complicated with sepsis were selected and categorized into the group with shock (n=88) and the group without shock (n=42). The general clinical data and laboratory indexes of the two groups were compared. The factors affecting the occurrence of septic shock were explored, and the correlation between serum sENG, BISAP, APACHEII, and SOFA scores was analyzed. The value of sENG and BISAP scores for diagnosis of SAP complicated with sepsis was assessed. Results APACHEII score, SOFA score, BISAP score, and serum sENG levels were higher in the group that developed septic shock. Increased BISAP score and elevated serum sENG level were independent risk factors for septic shock in patients with SAP. sENG level was positively correlated with BISAP score, APACHEII, and SOFA score in patients with SAP-complicated sepsis, and BISAP score was also positively correlated with APACHEII and SOFA score. sENG level and BISAP score had a predictive value for patients with SAP complicated with septic shock (AUC=0.723, 0.703), and the combination of the two had the highest value for the diagnosis of SAP complicated with septic shock (AUC=0.838). In addition, the AUC values of the two in predicting poor prognostic outcomes in patients with SAP complicated with sepsis were 0.757 and 0.706, respectively, and the AUC of the combination was 0.796. Conclusions Serum sENG and BISAP scores are predictors of septic shock in patients with SAP, and the combination of the two has a more powerful predictive effect and better evaluation significance.

Abstract Objectives Circular RNAs (circRNAs) are known to be associated with cardiovascular diseases. At present, an ideal biomarker for the early diagnosis of coronary heart disease (CHD) is still lacking. Methods We screened differentially expressed circRNAs (DEcircRNAs) in the peripheral blood monocytes (PBMCs) of patients with CHD, using the microarray technology in comparing the transcriptome. We identified upregulated and downregulated circRNAs. At the same time, we collected the patient clinical medical records and the PBMCs, the above results were analyzed and validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), using 374 patients. Results We identified 183 upregulated and 41 downregulated circRNAs. Among these DEcircRNAs, hsa_circ_0000745/hsa_circRNA_101996 was significantly upregulated in a cohort of 297 patients with CHD and 77 non-CHD controls. Among patients with CHD, hsa_circ_0000745/hsa_circRNA_101996 was significantly upregulated in the unstable angina pectoris (UAP) and acute myocardial infarction (AMI) subgroups compared to the stable angina pectoris (SAP) subgroup. By dividing hsa_circ_0000745/hsa_circRNA_101996 expression into quartiles, we observed that the highest hsa_circ_0000745/hsa_circRNA_101996 expression quartile was a risk factor for CHD compared to the lowest quartile (odds ratio [OR]: 2.709; 95 % confidence interval [CI]: 1.126–6.519, p=0.026), after adjusting for the traditional risk factors (age, sex, body mass index [BMI], smoking, alcohol, C-reactive protein [CRP], small and dense low-density lipoprotein [sdLDL] and lipoprotein-associated phospholipase A2 [LP-PLA2]). Conclusions These data suggest that upregulated hsa_circ_0000745/hsa_circRNA_101996 in PBMCs is a risk factor for CHD and could be used as a biomarker of CHD.

Abstract Objectives Epstein-Barr virus (EBV) can cause lymphoproliferative disorders (PTLD) in immunodeficiency individuals. The pathogenesis of EBV infection depends on its effective recognition and elimination. Our study investigated the effect of peripheral lymphocyte subsets (PLS) on the elimination of EBV. Methods A retrospective single-center study included 63 patients with 17 pediatric liver transplant recipients with EBV-induced PTLD (PTLD group) and 46 patients diagnosed with EBV-induced mononucleosis (IM group). Dynamic monitoring of PLS with EBV-DNA loads was performed. Results EBV-DNA replicated at a high level (5.2E3∼5.93E7 copies/mL in PBMC) before treatment in all patients in PTLD group. B lymphocytes were the main infected cells. After treatment with Rituximab, the EBV-DNA loads decreased below the lower limit of detection in 10 patients (PTLD-stable disease, PTLD-SD group), and the viral loads replicated at lower level in six patients (PTLD-partial response, PTLD-PR group). In the PTLD-SD group, the percentage of CD3+CD8+ T lymphocytes increased beyond the normal range with the ascending of EBV-DNA loads, then it decreased to the normal range accompanied by the clearance of EBV. In the PTLD-PR group, the CD3+CD8+ T lymphocytes kept in the normal range, while the EBV kept on replication. Conclusions The increased number of CD3+CD8+ T lymphocytes occurred in parallel with the decline in EBV-DNA loads, which is the most useful index in estimating the host capacity of immuno-surveillance against EBV.

Abstract Objectives Aged patients are often characterized by difficult blood sampling conditions. Smaller needle gauge (G) may be beneficial for venous access and reduced pain perception, however, potentially at the expense of lower blood quality for laboratory measurements. We systematically compared two blood collection sets with different outer but equal inner diameters; different needle tips, and retract mechanisms in aged patients (Safety-Lok™, 23G, SL vs. UltraTouch™ Push Button, 25G, UT-PB) regarding clinical aspects and laboratory measurements. Methods Clinical examination and questionnaires were used in an aged cohort (n=161, average age=81.6 years), to determine characteristics of venipuncture, the phlebotomist’s assessment of blood draw including level of difficulty, and patient’s pain perception with either one or both blood collection sets. Sample quality was evaluated by laboratory analytics considering 13 parameters. Results SL, UT-PB, or both were used in 89 (55 %), 72 (45 %) or 36 (22 %) patients. The handling of the blood collection sets was perceived slightly easier for UT-PB compared to SL by the phlebotomist (−30 %, p=0.038). There was no significant difference in other parameters of the phlebotomist’s assessment or patients’ perception of blood collection. There was no clinically relevant difference between both sets in any of the laboratory measurements, including potassium and hemolysis index. Conclusions Clinical use of the UT-PB compared with SL in aged patients was associated with slight advantages of UT-PB, e.g. in the handling comfort for the phlebotomist. Sample quality, especially regarding hemolysis, was identical between both blood collecting sets, making its use uncritical in difficult venous conditions commonly seen in elderly patients.

Abstract Objectives The rapid detection of carbapenemase-producing bacteria is clinically important for selecting appropriate antimicrobial therapy. Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was used to detect carbapenemase activity. Methods In this study, we evaluated the detection power of MBT STAR-Carba test on identifying carbapenemase-producing bacteria isolated in Kobe city, Japan, compared with that of the KBM CIM Tris II kit using the modified procedure parameters. The obtained results were expressed as normalized logRQ values indicating a measure of hydrolysis efficiency. Results The MBT STAR-Carba test rapidly detected not only major carbapenemases, such as IMP-1 and IMP-6 that are most prevalent in Japan, but also GES-type and OXA-51-like carbapenemases, which are difficult to detect by reaction with inhibitors or KBM CIM Tris II by extending the incubation time. Conclusions The MBT STAR-Carba test will be beneficial in rapid identification of carbapenemases in clinical settings and environmental investigations.

Abstract Platelet counting is a fundamental clinical test for diagnosing haemorrhagic diseases, coagulation abnormalities, and certain autoimmune disorders, and it also serves as a critical basis for decisions regarding platelet transfusion. Common automated methods for platelet counting include the international harmonization protocol (IHP) based on flow cytometry, CD61 immunoplatelet count (CD61-imm), impedance platelet count (PLT-I), hybrid platelet count (PLT-H), optical platelet count (PLT-O), and fluorescence platelet count (PLT-F). The IHP, based on flow cytometry, is recommended as the reference measurement procedure (RMP) by the Ministry of Health of the People’s Republic of China, the International Council for Standardization in Hematology (ICSH), and the International Society of Laboratory Hematology (ISLH) due to its superior precision and accuracy. Despite the significant improvements in efficiency and standardization brought about by automation, traditional blood smear microscopic examination (PLT-M) remains indispensable in specific scenarios, such as low platelet counts or abnormal platelet morphology, to ensure the accuracy and reliability of platelet counting results from automated methods.