Introduction: The aim of this study was to determine the performance of the total testing process of complete blood count (CBC) on two different instruments in an emergency setting of a county hospital, and to design an appropriate internal quality control plan.
Materials and method: Two models of Statistical Quality Control (SQC) were evaluated on Sysmex XT-1800i and Cell-Dyne Ruby: 3 levels of commercial blood every 8 hours (N=9) and an alternative model using 3 levels every 12 hours (N=6) as shift changes. Total Error (TE) was calculated using the formula: TE=Bias%+1.65xCV%; Sigma score was calculated using the formula: Sigma=[(TEa%–Bias%]/CV%. Values for coefficient of variation (CV%) and standard deviation (SD) were obtained from laboratory data and Bias% from proficiency testing. For the pre-analytical phase Sigma score was calculated, while for post-analytical phase the turnaround time (TAT) was assessed.
Results: TE for all directly measured parameters, for both instruments, had lower values than Total Error allowable (TEa). CV% for almost all parameters had lower values than CV% derived from biological variation except for platelets (PLT) at low level on Sysmex XT-1800i and red blood cells (RBC) on Cell-Dyne Ruby. Sigma score ranged from as low as 2 to 10. Sigma score for pre-analytical phase was 4.2 and turnaround time was 36 minutes on average.
Conclusions: Given the performances of the total testing process implemented for CBC in our laboratory, performing the internal control after every 50 samples/batch seems to fulfill both the Health Ministry Order (HMO) 1301/2007 and International Organization for Standardization ISO 15189:2013 recommendation. All quality instruments must work together to assure better patient results and every laboratory should design its own control plan that is appropriate for better quality achievement.
Introduction: Experimental acute cerebral ischemia quickly triggers circulating inflammatory cells, provoking infiltration of neutrophils and macrophages in the damaged brain region. N-3 polyunsaturated fatty acids alleviate the ischemic deterioration, however, their potential effect on bone marrow cell mobilization is less known.
Materials and methods: healthy male Wistar rats were submitted to intraperitoneal saline injection (n=10, sham Group), transient middle cerebral artery occlusion (tMCAO) and saline injection (n=10, placebo Group), tMCAO and highly purified fish-oil administration (n=10, T Group). At the two latter groups, twenty-four hours after tMCAO, MRI scans were performed to identify the ischemic regions; the eligible animals were sacrificed, the left parietal bones being removed and subjected to qualitative and quantitative histological and immunohistochemical analysis.
Results: The active hematopoietic surface was maximal at the T-Group, being significantly lower in the P- and S-Groups (p=0.006 and p= 0.017). The MPO positive surface increased significantly in the T-compared to the S-Group (22.57± 0.86 % vs. 18.87± 0.68%, p= 0.004). Arg1 expression was significantly higher (p=0.001), while iNOS expression was lower (p=0.004) in the T- than in the P-Group, but similar to the S-group. The Arg1/iNOS2 ratio was higher in the FO-treated than in the P-group (p<0.001).
Conclusion: the ischemic conditions triggered granulopoiesis and the increase of iNOS2 positive, type M1 macrophage in the rat bone marrow. Fish-oil treatment generated the expansion of overall hematopoietic surface along with normalization of iNOS2, up-regulating the potentially protective Arg1 positive M2 type macrophages and causing a significant shift in the M2/M1 ratio.
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.
Introduction: Pre-eclampsia is a pregnancy-specific disease characterized by hypertension after 20 weeks of gestation and proteinuria. It is a major cause of maternal and perinatal morbidity and mortality. The pathogenesis of pre-eclampsia is not completely understood. In our study we investigated if there is a potential link between cytomegalovirus infection and pre-eclampsia and if cytomegalovirus infection is the triggering factor of pre-eclampsia.
Material and methods: This study was carried out in a secondary care hospital between January 2014 and July 2015. We included two groups of pregnant women: 66 with pre-eclampsia and 62 without pre-eclampsia. Enzyme linked immunosorbent assay (ELISA) technique was performed to detect cytomegalovirus Immunoglobulin G levels in maternal serum.
Results: The p-value between median values of positive cytomegalovirus IgG in pre-eclamptic women and in controls was not significant (p:0.867). Odds ratio for cytomegalovirus IgG in pre-eclamptic group (OR:0.967; 95%CI:0.535-1.748) do not differ greatly from the value recorded in the control group (OR:1.036; 95%CI:0.571-1.880). The risk for preterm birth in pre-eclamptic women with cytomegalovirus IgG positive values (OR:1.009; 95%CI:0.329-3.090) was greater than those which had term delivery (OR:0.994; 95%CI:0.472-2.095) but the two values are not very high. We found a positive correlation coefficient (0.217) at a “p” value of 0.08 between white blood cells, but a negative correlation coefficient between the percentage of neutrophils and cytomegalovirus infection.
Conclusion: Cytomegalovirus infection may be associated with pre-eclampsia but it is unlikely to be the triggering factor of pre-eclampsia.
Background: Recommendations have been made, following the multicenter Surviving Sepsis Campaign study, to standardize the definition of severe sepsis with reference to several parameters such as haemodynamic stability, acid-base balance, bilirubin, creatinine, International Normalized Ratio (INR), urine output and pulmonary functional value of the ratio between arterial oxigen partial pressure and inspiratory oxigen concentration. Procalcitonin (PCT) is considered to be a gold standard biomarker for the inflammatory response, and recent studies have shown that it may help to discover whether a seriously ill person is developing sepsis. C-reactive protein (CRP) is also used as a marker of inflammation in the body, as its blood levels increase if there is any inflammation in the body. The aim of this study was to evaluate serum procalcitonin and C-reactive protein levels as diagnostic and prognostic biomarkers of severe sepsis.
Material and method: Sixty patients, diagnosed as being “septic”, were admitted to the intensive care unit (ICU). Based on laboratory results and clinical findings a diagnosis of “severe sepsis“ was made, and correlated with PCT and CRP values. The APACHE II, SAPS II and SOFA severity scores were calculated, analyzed and correlated with PCT and CRP.
Results: Fifty two patients (86.67%) presented with criteria for severe sepsis. Multivariate correlation analysis indicated a significant positive association between procalcitonin and all severity scores (APACHEII p<0.0001, SOFA p<0.0001, SAPS II p<0.0001). CRP proved to be significantly correlated only with the SAPS II score (p=0.0145). Mortality rate was high, with 48 patients (80%) dying. There was no significant correlation between the levels of the PCT and CRP biomarkers and severe sepsis (p=0.2059 for PCT, p=0.6059 for CRP).
Conclusions: The procalcitonin levels are highly correlated with the severity scores (APACHE II, SAPS II, SOFA) regularly used in ICUs and therefore can be used for determining the severity of the septic process. Quantitive procalcitonin and C-reactive protein analysis was not shown to be useful in diagnosing severe sepsis. However, PCT and CRP can be used to predict the fatal progression of the septic patient.
Background. The study aimed to evaluate the correlations of cognitive function with metabolic, nutritional, hormonal and immunologic parameters in patients with type 2 diabetes (T2D), in order to identify markers of cognitive impairment.
Material and methods. This cross-sectional study included 216 T2D patients and 23 healthy individuals (HC). The cognitive status was evaluated by the MoCA test. From HC and 145 T2D patients several parameters were also determined: C-peptide, vitamin B12, high-sensitivity CRP (by chemiluminescent immunometric assay), HbA1c, lipids, cortisol, TSH, Mg (by a Cobas 6000 analyzer), glucose (by glucose-oxidase method) and leptin and adiponectin (by ELISA method). Statistical significance was set at p < 0.05.
Results. There was a significant difference in the MoCA scores between HC and T2D groups (26.0(17.0-29.0) vs. 23.0(13.0- 31.0) points; p: 0.004). T2D patients with cognitive dysfunction were significantly older and less formally educated (p < 0.0001). Age negatively correlated with MoCA scores (-0.31; 95%CI:-0.42,-0.18; p < 0.0001). T2D patients had significantly lower visuospatial/executive (4.0(0.0-5.0) vs. 5.0(2.0-5.0) points; p: 0.04) and delayed recall scores (2.0(0.0- 5.0) vs. 3.0(1.0-5.0) points; p: 0.03) and lower serum Mg concentrations (0.81(0.12-0.99) vs. 0.92(0.41-1.35) mmol/l, p < 0.0001). Serum Mg levels positively correlated with MoCA scores (0.24, 95%CI: 0.07, 0.39; p: 0.003) and with visuospatial/ executive (0.30; 95%CI: 0.14, 0.45; p: 0.0002) and naming functions (0.18; 95%CI: 0.01, 0.34; p: 0.02).
Conclusions. Patients with T2D had significant cognitive impairment, with decrements in the visuospatial/executive and delayed recall domains. Younger age and higher education correlated with better cognitive function. Serum Mg levels correlated positively with overall cognitive function and with visuospatial/executive and naming domains.
Objectives: The primary aim of this study was to assess residual beta cell function at diagnosis of type 2 diabetes and identify accessible laboratory markers that best estimate it. The secondary objective was to evaluate the change in beta cell function 6 months after starting different therapeutical regimens. Materials and methods: Forty seven subjects were included in the study and each performed a 75-g oral glucose tolerance test (OGTT) at baseline and after 6 months. Metabolic and immunologic parameters were determined from fasting samples. According to the degree of metabolic decompensation, specific therapy was started: metformin, metformin plus gliclazide or insulin therapy (with/out metformin). Early and total beta cell function was evaluated by the disposition index (DI) calculated for 30 minutes and 120 minutes, respectively. Results: At diagnosis, fasting blood glucose (BG) and HbA1c varied largely (129-521 mg/dl and 5.5-14%, respectively). The DI30 and DI120 decreased with more severe glycemic decompensation. For both DI30 and DI120 significant negative correlations were found for glycemic markers (HbA1c, 2-hour BG and maximal BG amplitude) and positive correlation for 2- hour C peptide (p<0.0001 for all). HbA1c value of 7% discriminated an important decrease of DI30 and DI120. Insulin and combined therapy significantly improved DI120 at 6 months (p: 0.0062 and 0.01, respectively), while DI30 was improved only with insulin therapy (p: 0.0326). Conclusions: Beta cell function at onset correlated with HbA1c, 2-hour BG and C peptide during OGTT. Thus OGTT and HbA1c are pivotal for evaluation of beta cell function. Insulin therapy improved early and total insulin secretion at 6 months.
Introduction: Angiopoietin-2 (ANG-2) is a new biomarker whose blood-serum values increase in sepsis and its expression is elevated in line with the severity of the degree of inflammation. The aim of this study was to identify the diagnostic role of ANG-2 in patients with non-surgical sepsis addmitted to an intensive care unit.
Material and methods: This was a prospective randomized study including 74 patients admitted in the Clinic of Intensive Care of the County Clinical Emergency Hospital Tirgu Mure., divided into two groups: Group S: patients with sepsis (n=40, 54%) and Group C: control, without sepsis (n=34, 46%). ANG-2 levels were determined in both groups.
Results: From the Group S, 14 patients (35%) had positive haemocultures. ANG-2 values varied between 1 and 43 ng/mL, with an average of 6.0 ng/mL in patients without sepsis and 10.38 ng/mL in patients with sepsis (p=0.021). A positive correlation between ANG-2 and SAPS II, SOFA and APACHE II severity scores was demonstrated, as was a positive correlation between serum levels of ANG-2 and procalcitonine. ANG-2 had a 5.71% specificity and 74.36% sensitivity for diagnosis of sepsis.
Conclusions: ANG-2 serum levels were elevated in sepsis, being well correlated with PCT values and prognostic scores. ANG-2 should be considered as a useful biomarker for the diagnosis and the prognosis of this pathology.
Background: During the acute inflammatory process, the CXCL13 chemokine plays an important role in B cell recruitment within the central nervous system (CNS).
Objective: The objective of the study consisted of the evaluation of CXCL13 chemokine cerebral spinal fluid (CSF) and plasma levels in patients with acute infectious and non-infectious neurological diseases correlated with pleocytosis and CSF protein levels.
Material and method: This retrospective study was conducted over one year and included 72 patients. Thirty-eight patients (52.8%) suffering from infectious neurological disease, acute viral and bacterial meningitis, meningoencephalitis, and 34 patients (44.2%) diagnosed with non-infectious neurological diseases.
CXCL13 chemokine CSF and plasma levels were determined through the ELISA technique with the Human CXCL13/BLC/BCA-1 kit. CSF cell count, glucose and protein levels, along with anti-Borrelia burgdorferi antibodies were monitored using the ELISA technique.
Results: CXCL13 chemokine levels in the CSF of patients with acute infectious neurological diseases showed a median value of 23.07 pg/mL, which was significantly higher in comparison with the median value of 11.5 pg/mL of patients with noninfectious neurological diseases (p-0.03). CXCL13 median plasma concentration in patients with infectious neurological diseases was 108.1 pg/mL, in comparison with the second patient category, 50.7 pg/ml (p-0.001). We observed a statistically significant association between CXCL13 concentrations, CSF cell count and proteins. The higher the CXCL13 chemokine level, the more increased the cell count was.
Conclusions: CXCL13 levels in the CSF was significantly increased in patients with acute infectious neurological diseases compared with patients with non-infectious diseases. Moreover, CXCL13 chemokine concentration was significantly correlated with the number of cells and proteins in the CSF of patients suffering from neuroinfections.
Background: Twenty-four hours dynamic blood pressure (BP) behaviour displays dipper profile defined as nocturnal systolic BP (SBP) reduction>10% compared to daytime. Non-dipper profile, nocturnal absence of SBP fall, associates an increased cardiovascular risk. We investigated the concomitant association of inflammatory bio-markers - high-sensitivity- C-reactive protein (hs-CRP), Human Chitinase3-like1 (YKL-40) and autonomic nervous system (total brain-derived neurotrophic factor, BDNF) with respect to non-dipping blood pressure status.
Material and method: Using 24h automatic BP measurements, 80 known hypertensive patients were divided into two groups: dipper group included fifty-one dipper patients (age 55.6 ±13.5 years) and non-dipper group consisted of 29 non-dipper subjects (62.07±12.03 years). Serum levels of hs-CRP were evaluated with enhanced immunoturbidimetric assay. Plasma levels of YKL-40 were measured by commercial sandwich ELISA using microplate coated with a monoclonal antibody for CHI3L1. Plasma levels of total BDNF were determined using quantitative sandwich enzyme-linked immunoassay. Statistical analysis of obtained data was performed.
Results: In the non-dipper group, a significant positive association with age over 60 years, hs-CRP values above 1.90 mg/dl was observed along with increased mean values of YKL-40. Non-dipper status is independently and statistically significantly associated with elevated levels of hs-CRP (OR: 3.248, 95% CI: 1.022-10.317, p=0.046) in multivariate odds ratio analysis. No statistically significant positive association between a median total BDNF plasma level of 1430 pg/ml and the non-dipper hypertension profile was identified.
Conclusion: Our study demonstrated that patients over 60 years, in particular, have a higher probability to present a non-dipping pattern of hypertension. hs-CRP and YKL-40 values are more likely to increase in the non-dipping hypertensive patients, and hs-CRP values above 1.9 mg/dl can identify the presence of a non-dipper blood pressure profile.