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  • Author: Minodora Dobreanu x
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Abstract

Starting with the first issue of 2013, the Romanian Review of Laboratory Medicine has implemented a new editorial and publishing system. By this editorial, we try to clarify to all the readers and authors the major changes and their outcome in the journal’s evolution. Thus, we present details related to the current internal organization of the editorial board and the editorial workflow of the submitted manuscripts.

Abstract

Background: Trauma in its early stages leads to an acute inflammatory condition affecting all cellular lines. Neutrophil granulocytes make up the largest population of human white blood cells and are fundamental to the innate immune system. The objective of our pilot study was to evaluate neutrophil death and viability alterations in critically ill trauma patients in correlation with their clinical outcome.

Material and method: Critical ill trauma patients were enrolled in the study. In order to assess alterations in cellular death, blood samples were drawn using EDTA containing tubes and analyzed in the first twenty four hours after admission, then after forty eight and seventy two hours. Annexin V was used as a marker for apoptotic cells and propidium iodide for necrotic cells.

Results: The first two cases exhibited an increase in cellular viability by the second day as shown by a small increase in neutrophil apoptosis and a decrease in neutrophil necrosis. These patients progressed to a positive clinical outcome. The second two cases showed slight modifications in either physiological or pathological cellular death, and increasing levels of cellular necrosis. These patients progressed to a negative clinical outcome.

Conclusions: These cases suggest that neutrophil cell viability and death were associated with the patient’s clinical outcome.

Abstract

Objective: The aim of this study was to verify in our laboratory conditions the performance criteria of a commercial kit (PhagoburstTM, Glycotope Biotechnology) as described by the producers. We have also partially altered the use of the available kit by introducing a non-opsonized Candida albicans stimulus, in addition to the opsonized Escherichia coli stimulus provided by the manufacturer. Material and methods: The peripheral blood samples of 6 clinically healthy adults were tested in triplicate according to the manufacturer recommendations. The intraassay imprecision as well as the ranges of neutrophil and monocyte burst activation triggered by various stimuli were assessed. Results: The activation range of granulocytes and monocytes was similar to the one described by the producer in the presence of E. coli (granulocytes: 78.45-99.43% versus 99.6-99.95%, average %CV of 1.53% versus 0.1%, monocytes: 54.63-92.33% versus 81.80-96.67, average %CV 6.92% versus 1.1%). The leukocyte range of activation in the presence of non-opsonized C. albicans was comparable to the one triggered by the fMLP (N-formyl-methionyl-leucyl-phenylalanine) stimulus. Conclusion: The intra-assay precision obtained in our laboratory conditions, as well as the ranges of activated leukocytes, are comparable to the ones described by the producer when using E. coli as a stimulus. The present study shows that introducing an extra fungal stimulus for burst oxidation assessment could provide additional information regarding the non-specific cellular immune response, particularly in patients at risk for candidemia.

Abstract

Clinical laboratory tests ensure approximately 70% of the medical decisions, so that the time until the release of the results and its accuracy are critical for the diagnosis and the efficiency of the treatment [1]. Risk management involves both the anticipation of what could happen erroneous and the assessment of errors’ frequency as well as the consequences or the severity of the effects caused by it, and finally to decide what can be done in order to reduce the risk to an acceptable clinical level. For this reason, organizations should not see the risk management as a compliance issue, but as an integral part of the decision-making process. EP23-A is a guideline of CLSI that introduces the risk management principles in the clinical laboratory and encourages the laboratories to develop plans of risk management which are addressed to the risks of each laboratory. EP18-A2 proposes 2 techniques for identifying and controlling the errors in the laboratory: FMEA (Failure Mode and Effects Analysis) and FRACAS (Failure Reporting, Analysis and Corrective Action System). The European Committee of Experts and Management of Safety and Quality in Health Care proposed to use the quality indicators to identify the critical stages of each process, thus being possible to assess continuously the medical processes with the aim of identifying the errors when they occur. This review summarizes the principles of the risk management in the clinical laboratory, thus it can achieve its aims to report valid, accurate and reliable test results

Abstract

The aim of this study was to determine the rate of hemolyzed specimens sent to our laboratory for coagulation testing, assess the interference of hemolysis on coagulation for patients without anticoagulant therapy and to determine the reference intervals for PT, INR and aPTT for our laboratory in order to test our own limitations.

Methods: To determine the hemolysis rate, 1,689 specimens were evaluated on a visual scale and with the hemolysis icterus lipemia (HYL) test on Architect c4000 instrument. 125 blood samples collected from subjects without anticoagulant therapy were hemolyzed in vitro and the PT, INR and aPTT results were compared before and after hemolysis.To determine reference intervals (RI) for PT, INR and aPTT in our population, 125 apparently healthy human subjects (according to CLSI C28-A2) were enrolled and tests were performed on Sysmex CS 2000i, using Siemens reagents.

Results: Out of 1,689 samples, 9.46% were assessed as hemolyzed by the visual scale, while HYL test showed a 6.63% hemolysis rate. We found a shortening of 0.1s for PT, a diminution with 0.01 units for INR and a prolongation with 0.9s for aPTT from in vitro hemolyzed compared to non-lyzed samples. As to the reference intervals, we obtained in our laboratory versus reagents producer: for PT 9.8-13.9 s vs 9.8-12.1 s, and for aPTT 19.1-31.5s vs 23-31.9 s respectively; 28.38% more PT results and 13.44% more aPTT results were within range when we used local laboratory RI, compared to the manufacturer’s RI.

Conclusions: The rate of hemolyzed coagulation samples in our laboratory is higher than the rate found in the literature. Nevertheless, for patients without anticoagulant therapy hemolyzed samples should be processed. Using our own reference interval leads to a significant reduced number of abnormal results.

Abstract

Peripheral blood lymphocytes (PBL) are able to synthesize various cytokines that play key roles in the immune response and intercellular signaling. Since alterations in cytokine production and/or activity occur in many pathological processes, the study of cytokine synthetic capacity of PBL is a valuable tool for assessing the immune profile. In this paper, we aimed to investigate the variability of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) synthetic capacity of CD4+/CD8+ T-cells stimulated ex-vivo in healthy subjects, by means of a commercial intracellular cytokine staining (ICS) protocol. Peripheral blood mononuclear cells were isolated from 16 healthy subjects by Ficoll gradient centrifugation and activated ex-vivo with PMA/Ionomycin/Brefeldin-A for 4 hours. Activated PBL were surface-stained for CD3/CD4/CD8, fixed and permeabilized. ICS was performed using anti-human IL-2/TNF-α/IFN-γ and samples were analyzed on a BD-FACSAria-III flow cytometer. We recorded high post-isolation and post-activation mean viabilities: 82.1% and 82.4% respectively, p=0.84. Both CD4+/CD8+ subpopulations were found to partially produce each of the three cytokines, but in different proportions. On average, a significantly greater percentage of CD4+ cells was shown to produce IL-2 and TNF-α, compared with CD8+ cells (61.5%+/-5.8 vs. 25%+/-5.6 and 26.9%+/-11 vs. 7.5%+/-3.3 respectively, p---lt---0.0001 for both). Contrarily, IFN-γ was produced by a higher proportion of CD8+ cells (8.4%+/-3.9 vs. 6.8%+/-3.2, p=0.01). These results show that the employed ICS protocol elicits a satisfactory and consistent cytokine response from PBL of healthy subjects. The collected data may be used to outline a preliminary reference range for future studies on both healthy/pathological subjects.

Abstract

Background: Pediatric onco-hematology is not a frequently encountered medical specialty, and it influences everyday life, basic activities, and the immune system, mostly through psychosocial changes, which may affect every individual and their families differently. Anxiety is the most frequently encountered mental health disorder occurring during childhood and adolescence. The effect of stress and anxiety on the immune system is suggested by the fact that stress hormones elevate proinflammatory cytokines and subsequently lower the anti-inflammatory response.

Objective: Our main objective was to analyze the relationship between anxiety disturbance and cytokine levels in oncologic pediatric patients from Târgu Mureș in order to answer the following question: does anxiety influence immunity?

Material and methods: After testing pediatric oncology patients from the Pediatrics Clinic no. 2 of Târgu Mureș, Romania with the SCARED child test, we took blood samples from each participant. IL-6, IL-10, IL-1β, IL-12p40 and TNF-α levels were evaluated with a Human Cytokine Magnetic Panel using the xMAP technique on Flexmap 3D platform (Luminex Corporation, Austin, USA). C-reactive protein levels were determined with the BN Pro Spec nephelometer with CardioPhase hsCRP (Siemens Healthcare Diagnostics, GmbH, Marburg, Germany) reagent.

Results: The 46 pediatric oncology patients had 6 main diagnostic groups, the most frequent pathology was acute leukemia (58.7%) followed by malignant solid tumors (21.74%) and lymphomas (6.52%). In the anxious group (45.65%) we observed 4 of the 5 studied anxiety types: panic disorder, separation, social, and generalized anxiety. We measured the cytokine levels of all the participants from the two main groups: anxious/non-anxious. Statistical analysis (linear regression) showed statistically significant positive correlations in the anxious group related to the IL-1β and IL-6, a moderate/weak correlation related to IL-12p40, as well as a negative moderate correlation between IL-10 values in the anxious group and a positive trend in the non-anxious group.

Conclusions: Psycho-oncology is a relatively young specialty with few studies in the last two decades. IL-1β, IL-6, and TNF-α present high levels in anxious patients, while IL-10 and IL-12p40 have low serum levels in mental disorders. C-reactive protein levels are not influenced by anxiety.

Abstract

Morphological and immuno- flow cytometry assisted analysis of peripheral blood and bone marrow are mandatory investigations in the diagnosis of acute leukemia. Cytology and immunophenotyping complement each other primarily because they have as common object malignant cell phenotype as a whole. The aim of our study was to analyze correlations between cytology and immunophenotyping on a group of patients investigated for acute myeloid leukemia. In our study the degree of correlation between blast percentage determined by cytology and immunophenotyping was low (r=0.049). The degree of correlation between myeloperoxidase positivity in cytochemistry and immunophenotyping was also low, with better results for cytochemistry. Expression of immunophenotypic markers was consistent with the composition of our group regarding French-American-British classes, except for HLA-DR (49.0%), TdT (3.77%), CD14 (5.66%), CD15 (5.66%). We also discuss the importance of interpreting with caution positivity for erythroid and megakaryocytic markers and differential diagnosis of cases simultaneously expressing CD7 and CD56. In conclusion, interpretation of immunophenotyping by flow citometry, done in close conjunction with morphology, is mandatory to facilitate the use of optimized sample processing methods and of standardized panels, for both appropriate diagnosis and follow-up.