Volume 46, Issue 1

Human serum paraoxonase 1 (PON1) appears to play an important role in the development of a large variety of diseases with an inflammatory component including heart disease, diabetes, rheumatic diseases, neurological diseases and cancer. As such PON1 research is rapidly expanding into new biomedical fields. Unfortunately, this rapid expansion has resulted in a number of problems due to poor experimental design and the spreading of misconceptions in the literature. This review seeks to describe the basic properties of PON1 and the problems and misconceptions that have arisen.

The basic principle in the clinical use of Platelet Rich Plasma (PRP) is to increase the innate repairability of the human body and accelerate recovery after injury. PRP, a source of natural growth factors, emerges as a treatment method to reduce the side effects related to radiotherapy. PRP exerts its radioprotector effect by stimulating angiogenesis, epithelialization, cell differentiation and extracellular matrix formation in aged and poorly regenerated tissues. Additional studies are needed to confirm the promising effect of PRP in beam therapy.

Objectives Preanalytical phase quality, which begins with a request for testing and continues with patient preparation, sample collection, transport, processing and storage, is essential for coagulation tests. The Turkish Biochemical Society Preanalytical Phase Working Group created a survey to evaluate the preanalytical phase practices for coagulation laboratories in our country. Methods The survey consisted of a total of 26 questions and included almost all steps of the preanalytical phase. Results Fifty-four laboratory specialists have participated in the study. The survey results showed that participants have different practices for most stages of the preanalytical phase for coagulation tests. Conclusion According to the survey results, a national guideline may help standardisation of the preanalytical phase for coagulation tests in our country and increasing training in this respect would contribute to achieving accurate test results.

Objectives We aimed to evaluate the ability of lymphocyte-C-reactive protein ratio (LCR) to discriminate between different levels of severity of COVID-19 disease. Methods This retrospective observational single-center study was performed on 61 confirmed (PCR positive) COVID-19 patients between March and June 2020. The study population was separated into three groups: mild/moderate (n=24), severe (n=25) and critically ill (n=12). The optimal cut-off values of the LCR and neutrophil-to-lymphocyte ratio (NLR) in discriminating between patients with different severity levels were calculated by applying the receiver operating curve (ROC) analysis. Results At baseline, the LCR decreased significantly across the three severity groups (mild/moderate > severe > critically ill). ROC analysis showed that a mean LCR of 43.21 was the cut-off value which best discriminated patients with the critically ill disease from severe patients (sensitivity: 84% and specificity: 69%). The discriminative performance of LCR (ROC AUC 0.820) was better than that of NLR (0.751) in this regard. LCR, unlike NLR was able to distinguish severe patients from mild/moderate patients, with a cut off value of 458.19 (sensitivity: 80% and specificity: 45%). Conclusion LCR was observed to be able to distinguish COVID-19 infected patients of different severity (mild/moderate, severe and critically ill) and was superior to NLR in this regard.

Objectives We performed a verification study of the Sysmex XN-3100 hematology analyzer in comparison with the XE-2100 according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI) and the International Council for Standardization in Hematology (ICSH). Materials and methods Blood samples and quality control materials were used for precision. For comparison, we used the current XE-2100 as the comparative method and analyzed 540 blood samples. The Passing-Bablok and Bland-Altman tests were performed according to the CLSI EP09-A3 and a carryover study was performed according to the CLSI H26-A2 guidelines. The flagging performance of the two analyzers was compared, using two experienced laboratory technicians as the reference method. Results The Sysmex XN-3100 demonstrated high levels of precision for most parameters. For the comparison analysis, all parameters, except for MCHC, monocytes and basophils were within the systematic error limits of desirable biological variability criterion (SeDBV). The carryover was less than 0.4% for all parameters. The flagging performance of the XN-3100 was satisfactory and the overall efficiency was high. Conclusions The XN-3100 not only showed a strong correlation and agreement with the XE-2100 but also displayed a comparable analytical sensitivity, and increased specificity, which may result in an improved turnaround time and throughpu.

Objective Hemoglobinopathies are a common public health problem in Turkey. In the screening of these disorders in population, cation-exchange high performance liquid chromatography (HPLC) is accepted as the gold standard method. In this study, the aim was to assess four different HPLC devices used in hemoglobinopathy screening. Materials and methods A total of 58 blood samples were analyzed with four different HPLC methods (Bio-Rad variant II, Agilent 1100, Tosoh G8 and Trinity Ultra2 trademarks). Results The comparison study demonstrated a good correlation between the results of each HPLC analyzer and the reference value obtained by averaging all the HbA 2 results belonging to the methods tested in the study [ (Tosoh G8 (r=0.988), Bio-Rad variant II (r=0.993), Agilent 1100 (r=0.98) and Trinity Ultra2 (r=0.992) ]. HbA 2 determination in the presence of HbE was interfered in both Bio-Rad variant II and Tosoh G8. Conclusion The analyzers were found to have compatible HbA 2 results but with accompanying different degrees of proportional and systematic biases. HPLC analyzers may be affected by different hemoglobin variants at different HbA 2 concentrations, which is an important point to take into consideration during the evaluation of HbA 2 results in thalassemia screening.

Objectives This study investigated the diagnostic power of reticulocyte hemoglobin equivalent (Ret-He) in the differential diagnosis of hypochromic microcytic anemia to differentiate iron deficiency anemia (IDA) and thalassemia trait (TT) based on the traditionally used erythrocyte index and formulas. Methods Twenty-six children with iron deficiency (ID), 26 with IDA, 33 with β-TT, 41 healthy children were assessed. Complete blood count parameters, Ret-He, immature reticulocyte fraction (IRF), low-fluorescence ratio (LFR), Mentzer’s indexes (MI) were evaluated. The diagnostic power of Ret-He in distinguishing between IDA and β-TT was investigated using ROC analysis. Results Ret-He levels were (median(Q1-Q3)) 20.6(19.7–21.5) pg in β-TT, 16.1(13.1–20) pg in IDA, 29.7(27.2–30.7) pg in ID, 30.5(29.8–31.7) pg in healthy controls. Based on ROC analysis, diagnostic power for distinguishing between IDA and β-TT was determined as RBC>MI>Ret-He>RDW>LFR>IRF. The highest sensitivity and specificity for differential diagnosis was obtained when the Ret-He cut-off value was 18.2pg. The AUC (95%CI) value was calculated as 0.765(0.637–0.866), and a statistically significant difference was found between groups (p<0.0006). Conclusions In patients with hypochromic microcytic anemia, Ret-He≤18.2pg combined with RBC≤5.3x10 6 /L and MI>10.42 can be safely used to distinguish IDA from β-TT. In particular, patients with low Ret-He who don’t respond to iron therapy should be examined for β-TT.

Background Sickle cell anemia (HbSS) is a hereditary blood disease that affects millions of people worldwide. Increased the HbF levels affects the clinical course of HbSS positively. The aim of this study was to investigate the association between miR-144-3p and HbSS. Materials and methods In this study 84 cases (44 HbSS/HbS combination and 40 HbAA) were performed. The expression of miR-144-3p was determined by RT-PCR. Statistical analysis was performed by the Mann-Whitney U test (SPSS 20.00 for Windows and p<0.005). Results The miR-144-3p expression levels were higher in the HbSS cases (p≤0.001). Additionally, it was determined that the expression of miR-144-3p was higher in the cases with HbF<3 (p=0.043). Discussion In our study, the increase in the miR-144-3p levels in low HbF levels may be associated with the severity of the disease. Conclusion Considering these results, suppressing miR-144-3p may be considered as a new treatment option in HbSS.

Objectives It has been known that some laboratory tests showed diurnal variation and were affected by fasting-satiety status. We aimed to examine erythrocyte sedimentation rate (ESR) for both pre-analytical factors. Methods Blood samples from 12 volunteers were taken and studied with the Westergren method. Fasting blood samples taken at 9:00 am were accepted as basal. Samples were taken at 10:00, 11:00, 12:00 were compared with baseline to evaluate fasting-satiety status and samples taken at 12:00, 15:00, 18:00, and 24:00 to evaluate diurnal variation. Results ESR was found to be the lowest at 09:00 and 15:00 (Median=5.5 mm/h). The 10:00, 11:00, and 12:00 results for assessing fasting-satiety status on ESR were not different than the baseline (6.2, 6.0, and 7.8 mm/h, respectively). The rates at 18:00 and 24:00 were not found to be different than the baseline (5.7 and 6.6 mm/h). Conclusions We didn’t find that ESR had diurnal variation, and it wasn’t affected by fasting-satiety status. Although it is known that routine biochemistry tests should be performed from the blood sample taken in the morning in case of starvation, blood sampling can be done for ESR during the day if necessary.

Background Multiple myeloma (MM), a malignancy of plasma cells, is the second most prevalent hematological cancer. Bortezomib is the most effective chemotherapeutic drug used in treatment. However, drug-resistance prevents success of chemotherapy. One of the factors causing drug-resistance is dysfunction of apoptotic-pathways. This study aimed to evaluate the relationship between expression levels of Bcl-2, Bax, caspase-3 and p-53 genes involved in apoptosis and the development of bortezomib-resistance in MM cell lines. Materials and methods Multiple myeloma KMS20 (bortezomib-resistant) and KMS28 (bortezomib-sensitive) cell lines were used. 3-[4,5-Dimethylthiazol-2-yl] 1-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine IC 50 values of bortezomib. RNAs were isolated from bortezomib-treated cell lines, followed by cDNA synthesis. Expression levels of the genes were analyzed by using q-Realtime-PCR. Results As a result, Bcl-2/Bax ratio was higher in KMS20 (resistant) cells than in KMS28 (sensitive) cells. Expression of caspase-3 decreased in KMS20-cells, whereas increased in KMS28-cells. The results indicate that apoptosis was suppressed in resistant cells. Conclusion These findings will enable us to understand the molecular mechanisms leading to drug-resistance in MM cells and to develop new methods to prevent the resistance. Consequently, preventing the development of bortezomib resistance by eliminating the factors which suppress apoptosis may be a new hope for MM treatment.

Objectives To demonstrate immature granulocyte (IG) count and delta neutrophil index (DNI) values (novel potential predictive marker for neonatal sepsis) for neonates. Methods This prospective controlled clinical study was consisted of 208 patients (77 in the study group and 131 in the control group) who were delivered between January 2016 and January 2018 at the Hacettepe University Neonatal Intensive Care Unit in Ankara, Turkey. In this study, we evaluated value of DNI in diagnosing neonatal sepsis by comparing the DNI values in culture positive septic neonates with healthy neonates. Results In our study, the median interquartile range (IQR = 25–75%) DNI was 0.1% (0.0–0.3%) in the control group and 1.5% (1.0–2.45%) in the sepsis group (p < 0.05). In our ROC curve analysis, the cut-off value for the DNI as a sepsis marker was 0.65%, with 96.2% specificity and 97.4% sensitivity. Those patients with gram-negative isolates had significantly higher DNI and IG counts when compared to those patients with gram-positive bacteria (p < 0.05). Conclusions Our findings indicated that the DNI counts are significant diagnostic biomarkers for neonatal sepsis. They may also have utility in determining the sepsis etiology (differentiating between gram-positive and gram-negative agents).

Objective Coagulation tests are sensitive to pre-analytical variables. The aim of our study is to identify the effect of transportation and freeze-thaw status on for Factor VIII, Factor IX, Anti-thrombin III, Protein S, Protein C, Prothrombin time (PT) and Activated partial thromboplastin time (aPTT). Materials and methods The study was performed on 102 plasma samples obtained from 34 healthy volunteers. The samples were divided into three groups. Group A was analyzed whereas group B, C were frozen at −20°C. After 24 h, group B and C were transported for 2 h. Following the transfer, group B was analyzed and C was frozen at −20°C. After 24 h, group C was analyzed. Analyses of samples were performed in Thrombolyzer-XRM for PT, aPTT, Factor VIII, Factor IX, Anti-thrombin III, Protein C and Protein S. Results There were significant variations for PT, aPTT, Protein S, Factor VIII and Factor IX for group A&B and A&C comparisons in different stability criteria approaches. In significant change limit and percentage change calculations Protein S, Factor VIII and IX showed significant differences. For acceptable change limit approach, aPTT and Factor IX showed significant changes. Conclusion Laboratories should take precautions for transportation and freeze-thaw cycles to prevent inaccurate results.

Objectives The purpose of this retrospective study was to investigate the relationship between 25 OH vitamin D (25[OH]D) deficiency and iron-deficient anaemia (IDA) in the pediatric population. This was aimed to provide a better insight to IDA follow-up and treatment. Methods The data of 120 patients diagnosed with IDA and 125 healthy pediatric patients were analyzed retrospectively. Serum vitamin B 12 , Folate and 25(OH)D levels, between IDA and healthy groups were evaluated. The relationship between vitamins levels and IDA parameters were examined. Logistic regression analysis was used to assess whether 25(OH)D deficiency levels were an independent risk factor for diagnosing IDA. Results In the comparison of vitamins levels between groups, only mean serum 25(OH)D levels were found to be statistically significantly (p=0.000) lower (13.00 ± 2.50 ng/mL) in the group with IDA compared to the healthy group (25.98 ± 3.66 ng/mL). There were strong positive correlations between 25(OH)D deficiency levels and IDA. The deficiency of 25(OH)D levels was not found to be an independent risk factor for IDA (ORs: 0.958, 95%CI: 0.917–1.000). Conclusions Although current results confirm the association between 25(0H)D deficiency and IDA in pediatric patients, they indicate that there was no independent risk factor for IDA.

Objectives Hemoglobinopathies are inherited transition blood diseases associated with globin chains of the hemoglobin. However many mutations have been defined, there may be many of them not defined yet. We here report the first case of those mutations, named Hb G-Norfolk in Turkey. Case presentation A 15 years-old male patient with erythrocytosis was referred to our laboratory for the evaluation of hemoglobinopathy. In chromatographic analysis, an unidentified peak was observed. A similar identification for variant Hb could not be obtained from High-Performance Liquid Chromatography (HPLC) analyzer’s data library. No definitive diagnosis could also be made by different analyzer. Family screening and molecular genetic DNA sequence analysis were carried out. Conclusions Although there were not found any beta gene mutation of neither the patient nor his family, analyses of alpha genes A1 and A2 were performed and abnormal hemoglobines were detected for all of them. This change in the HbA2 gene was at codon85 GAC>AAC (Asp>Asn) in the heterozygous state, known as Hb G-Norfolk [HbA2:c256G>A p.Asp85Asn] based on HbVar database. Abnormal Hb bands detected by HPLC with clinical findings such as erythrocytosis or cyanosis should be investigated by sequence analysis to corroborate alpha and/or beta-globin gene mutations for avoiding misdiagnosis and misinterpretation.