ISSN  2587-2362  |  E-ISSN  2618-642X
International Journal of Medical Biochemistry - Int J Med Biochem : 4 (3)
Volume: 4  Issue: 3 - 2021
1.Predictive value of immature granulocyte count and other inflammatory parameters for disease severity in COVID‐19 patients
Murat Alisik, Utku Gorkem Erdogan, Merve Ates, Mehmet Alp Sert, Ozgur Mehmet Yis, Guler Bugdayci
doi: 10.14744/ijmb.2021.03164  Pages 143 - 149
INTRODUCTION: This study was designed to compare the immature granulocyte (IG) count, IG-to-lymphocyte ratio (IGLR), complete blood count (CBC) values, and inflammatory parameters of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), C-reactive protein (CRP), ferritin level, and CRP-to-albumin ratio (CAR) measured at hospital admission in patients with coronavirus disease 2019 (COVID-19) and non-COVID-19 patients and to compare these parameters between subgroups according to disease severity. In addition, these parameters were evaluated for predictive value related to the severity of COVID‐19.
METHODS: The data of adult patients admitted with a suspected COVID-19 infection confirmed with real-time polymerase chain reaction testing of nasal and pharyngeal swab specimens were included in this retrospective study. Outpatient COVID-19-positive patients were enrolled in the mild group, hospitalized patients were classified in the moderate group, and patients admitted to the intensive care unit were categorized in the severe group.
RESULTS: A total of 1213 COVID-19-positive patients and 1034 COVID-19-negative patients were included in the study. The IGLR, NLR, PLR, CRP, CAR, and ferritin levels were significantly higher, and the leukocyte, IG, neutrophil, lymphocyte, monocyte, basophil, and eosinophil levels were significantly lower in the COVID-19-positive group than the COVID-19-negative group (p<0.05 for all). The severe group had higher median IG, IGLR, neutrophil, NLR, PLR levels than the mild, moderate, and COVID-19-negative groups (p<0.05 for all). Receiver operating characteristic analysis revealed an area under the curve value for IGLR, CAR, CRP, IG, NLR, and ferritin of 0.868, 0.860, 0.834, 0.848, 0.845, 0.841, and 0.827, respectively, which differentiated severe COVID-19 patients from mild and moderate COVID-19 patients.
DISCUSSION AND CONCLUSION: The results suggest that the IGLR may be useful to distinguish severe COVID-19 patients at the time of admission. Further exploration is warranted to fully determine the potential value of the IGLR in disease monitoring.

2.Reference change value and measurement uncertainty of CRP, ferritin, procalcitonin, troponin I, and BNP tests in COVID-19 monitoring: How much change is significant?
Nergiz Zorbozan, Orçun Zorbozan
doi: 10.14744/ijmb.2021.30602  Pages 150 - 156
INTRODUCTION: This study was designed to determine the point of clinically significant change in consecutive measurements of high concentrations of C reactive protein (CRP), ferritin, procalcitonin, troponin I, and brain natriuretic peptide (BNP) in patients diagnosed with COVID-19.
METHODS: The reference change value (RCV) for each analyte was estimated using the following formula: RCV=21/2×Z×(CVa2+CVi2)1/2. The combined RCV was calculated using the mean CV value determined using different concentrations. Measurement uncertainty (MU) was estimated according to NordTest guideline. The root-mean-square (RMS) bias and nominal uncertainty values (uCref) were calculated. Standard uncertainty (ubias) and combined standard uncertainty (u) were calculated using the formula RMSbias²+uCref² and √(uRW²+ubias²)/2, respectively. Expanded uncertainty (U) was obtained by multiplying u by a coverage factor.
RESULTS: The lowest RCV values of CRP, ferritin, procalcitonin, BNP, and troponin I tests were 94.57%, 37.16%, 40.55%, 42.01%, and 28.99%, respectively. The combined RCV value was 94.6%, 38.31%, 41.57%, 42.35%, and 29.36% for CRP, ferritin, procalcitonin, troponin I, and BNP, respectively. The U of the CRP test was between 4.70%, and 5.49%. The lowest MU values of ferritin, troponin I, and BNP tests were 48.45%, 48.24%, and 53%, respectively.
DISCUSSION AND CONCLUSION: Providing the RCV and MU values with the test results in laboratory reports of these important biomarkers, which are frequently seen in high concentrations in COVID-19 patients, would offer valuable information for more accurate clinical interpretation.

3.Evaluation of vitamin D and calcium mineral metabolism in patients with chronic urticaria
Saadet Kader, Selami Aykut Temiz, Turan Akdağ, Ali Ünlü
doi: 10.14744/ijmb.2021.08760  Pages 157 - 160
INTRODUCTION: Chronic urticaria (CU) is a skin disease characterized by the spontaneous occurrence of itchy, swollen welts lasting 6 weeks or more. The aim of this study was to assess the potential influence of vitamin D and calcium mineral metabolism in patients with CU.
METHODS: Patients over the age of 18 who presented at a dermatology outpatient clinic between November 2020 and February 2021 and diagnosed with CU were included in the study. Volunteers with no disease were selected as a control group. The level of 25-hydroxy vitamin D and calcium metabolism parameters (calcium [Ca], phosphorus [P], parathyroid [PTH] hormone) were measured in both groups and compared.
RESULTS: The mean age of the participants was 35.01±11.96 years in the urticaria group and 34.89±11.75 years in the control group (p>0.05). The mean serum Ca level was 9.14±0.75 mg/dL in the urticaria group and 9.46±0.43 mg/dL in the control group (p=0.003). The mean serum P level was 3.3±0.71 mg/dL in the urticaria group and 3.3±0.56 mg/ dL in the control group (p=0.76). The mean serum level of PTH was 46.28±16.29 pg/mL in the urticaria group and 41.95±14.09 pg/mL in the control group (p=0.091). The mean serum level of vitamin D in the CU and the control group was as 13.8±5.67 ng/mL and 18.12±8.42 ng/mL, respectively (p<0.001). The 7-day Urticaria Activity Score (UAS7) was 19.99±8.61 in the patients with urticaria. There was a negative correlation between the UAS7 score and the vitamin D level (p=0.001). The mean duration of disease was 20±15.22 months. No correlation was seen between disease duration and the vitamin D level (p=0.49).
DISCUSSION AND CONCLUSION: The study findings indicated that the Ca and vitamin D levels in patients with CU were statistically significantly lower than those of the control group. However, additional studies are needed to further explore the possible contribution of this analysis to the diagnosis and treatment of urticaria.

4.Saline replacement as a practical solution to matrix interference effect when a leukocyte differential count cannot be measured in chemotherapy patient samples
Hatice Bozkurt Yavuz, Hüseyin Yaman, Mustafa Tat, Osman Akıdan, Asım Örem, Süleyman Caner Karahan, Yüksel Aliyazıcıoğlu
doi: 10.14744/ijmb.2021.09825  Pages 161 - 165
INTRODUCTION: An automated complete blood cell count (hematology) analyzer cannot measure the white blood cell (WBC) differential of some patient sample, notably those of chemotherapy patients. This is often due to the presence of atypical cells in the sample or matrix changes caused by drugs. A technique to use isotonic sodium chloride solution (0.9% saline) as a replacement for plasma has previously been described. The aim of this study was to evaluate to use of saline replacement as a means to resolve the matrix interference effect and achieve a reliable WBC differential count for chemotherapy patients.
METHODS: Samples of 29 chemotherapy patients whose WBC differential count could not be calculated using a Beckman Coulter LH-780 hematology autoanalyzer (Beckman Coulter, Inc., Brea, CA, USA) were evaluated. A peripheral blood smear was performed and the saline replacement technique was applied and the samples were then re-analyzed using the same autoanalyzer. The WBC count and differential count of WBC of a peripheral smear and a saline-replacement sample were compared.
RESULTS: There was no statistically significant difference between the peripheral blood smear and saline replacement autoanalyzer measurements of the WBC differential count and percent of WBC. Strong relationships in WBC (r=0.99), neutrophil (r=0.98), lymphocyte (r=0.98), monocyte (r=0.91), and eosinophil (r=0.77) counts were observed in a comparison of the peripheral blood smear and saline replacement measurements.
DISCUSSION AND CONCLUSION: A saline replacement technique may be a practical solution to resolve the difficulty of the matrix interference effect seen in chemotherapy patient samples and provide a WBC differential count.

5.Distribution of thalassemia trait in Balikesir Province according to trait type and age group
Uğur Erçin
doi: 10.14744/ijmb.2021.20982  Pages 166 - 171
INTRODUCTION: The aim of this study was to determine the prevalence of the thalassemia disease group in Balikesir Province, Turkey.
METHODS: Blood samples provided by 11,558 individuals (5675 males and 5883 females) aged 3-90 years between May 15, 2018 and September 30, 2019 for laboratory analysis at the Balikesir Provincial Public Health Laboratory were included in the study. Hemoglobin chain analyses were carried out using the high-performance liquid chromatography (HPLC) method. The data obtained were analyzed retrospectively.
RESULTS: The level of hemoglobin (Hb) A2 was >3.5% in 591 (5.11%) of the total study group and these individuals were identified as β-thalassemia carriers. The prevalence of the β-thalassemia trait was 5.76% in females and 4.44% in males. A total of 792 cases (446 female and 346 male) had a result outside the normal range: 74.6% were identified as thalassemia carriers, 9.4% had isolated low Hb A2, 12.3% had isolated Hb F elevation, and 3.8% had total abnormal hemoglobin values.
DISCUSSION AND CONCLUSION: The prevalence of β-thalassemia trait (5.11%) in the study group was extrapolated for the general population of Balikesir (2019 population: 1,228.620 ) and it was estimated that there were 62,782 potential carriers in the province.

6.Evaluation of the Vision C erythrocyte sedimentation rate analyzer
Fevziye Burcu Şirin, İlter İlhan, Halil İbrahim Büyükbayram, Duygu Kumbul Doguc
doi: 10.14744/ijmb.2021.52244  Pages 172 - 177
INTRODUCTION: The International Council for Standardization in Haematology recommends that new methods and instruments used in medical laboratories be thoroughly tested with an analytical performance evaluation prior to implementation. This study is an assessment of the analytical performance characteristics of the Vision C analyzer (Shenzhen YHLO Biotech Co., Ltd., Shenzhen, China), which uses a modified Westergren method to measure erythrocyte sedimentation rate (ESR) and a comparison with the reference Westergren method.
METHODS: The analytical performance of the Vision C was evaluated according to intra-run/inter-run precision, stability, and compatibility with the gold standard Westergren method. In all, 173 patient samples were studied using both methods. The Westergren method levels were used to form 3 groups: <40, 40-80, and >80 mm/hour.
RESULTS: The intra-run precision ranged from 4.93% to 18.18% for 5 samples at different levels. Inter-run precision for the normal level was 7.03% and 2.94% for the pathological level. Both the room temperature and refrigerated samples were stable at 4 hours. The overall bias between the Westergren method and the Vision C analyzer was -5.23 mm/hour (confidence interval [CI]: -6.66 to -3.79), the correlation coefficient was 0.948, and the Passing-Bablok regression equation was y=2.073+0.805x. The bias of the Vision C for ESR values <40 mm/hour was -0.885 (CI: -2.027 to 0.258), -9.23 (CI: -11.853 to -6.613) for values 40-80 mm/hour, and -17.26 (Cl: -21.306 to -13.227) for values >80 mm/hour.
DISCUSSION AND CONCLUSION: The Vision C analyzer using a modified Westergren method met the precision and sample stability criteria, but the results were not fully compatible with the Westergren method at all ESR levels, and the difference was significant at high ESR levels.

7.Can immature granulocyte count and hemogram indices be good predictors of urinary tract infection in children?
Said İncir, Mehmet Taşdemir, Kerim Erhan Palaoglu, Hatice Kant Calti, Arzu Baygül, Ilmay Bilge
doi: 10.14744/ijmb.2021.52523  Pages 178 - 184
INTRODUCTION: A left shift in a complete blood count (CBC) indicates the presence of immature granulocytes (IGs), and an elevated count is associated with a bacterial infection. The aim of this study was to evaluate the role of the IG count and inflammatory CBC parameters and indices in children with a urinary tract infection (UTI).
METHODS: In this retrospective study, 55 children with a UTI (patient group) and 47 children with no infection matched for age and sex (control group) were enrolled. The symptoms, clinical findings, CBC parameters, and platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), systemic immune-inflammatory index (SII), and urinalysis findings were recorded. The C-reactive protein (CRP) level, urine culture results, and radiological data for the UTI group were also noted and analyzed.
RESULTS: The IG count; leukocyte, platelet, and neutrophil counts; and the CRP value were higher in the patient group than in the control group (p=0.001, 0.001, 0.035, 0.001, and <0.001, respectively). The NLR, PLR, SII, and mean platelet volume were not significantly different between the groups. The median hemoglobin level was significantly lower in the patient group than in the control group (10.9 g/dL vs. 12.2 g/dL; p=0.004). The area under the curve values for CRP, and counts of IG, leukocytes, and neutrophils were 0.851 (95% confidence interval [CI]: 0.761-0.917), 0.736 (95% CI: 0.633-0.823), 0.714 (95% CI: 0.609-0.804), and 0.670 (95% CI: 0.564-0.765), respectively.
DISCUSSION AND CONCLUSION: The IG count is an easily obtained measurement that can be utilized alongside the CRP value and other indicators to create better predictive value in the management of UTI.

8.Examination of fasting and postprandial dynamic thiol disulfide homeostasis and oxidative stress
Ahmet Rıfat Balık, Betül Özbek İpteç, Reyhan Ersoy, Gulsen Yilmaz, Ozcan Erel, Cemile Biçer
doi: 10.14744/ijmb.2021.53825  Pages 185 - 191
INTRODUCTION: Oxidative stress has a central role in metabolic disorders associated with high-carbohydrate, animal-based-protein diets, and excessive fat consumption. However, the molecular mechanisms of nutrition-mediated oxidative stress are complex and not fully understood. Dynamic thiol-disulfide homeostasis (DTDH) is the regulation of a balance between thiols and their oxidized forms, and includes the reversal of thiol oxidation in proteins. This study was an evaluation of DTDH and postprandial oxidative stress.
METHODS: A total of 86 participants (43 males and 43 females), were included in the study. Fasting and non-fasting blood samples were collected and the native thiol, total thiol, and disulfide parameters were analyzed. Statistical analyses were performed using IBM SPSS Statistics for Windows, Version 22.0 (IBM Corp., Armonk, NY, USA).
RESULTS: The findings indicated that while the native thiol values were significantly lower in the postprandial samples, the disulfide values were significantly higher. There was no significant difference in the total thiol values.
DISCUSSION AND CONCLUSION: Examination of the DTDH revealed that the oxidative stress level increased following food intake. Protein thiols involved in antioxidant defense were oxidized and transformed into disulfides.

9.Development and optimization of a new aptamer-based method for P-selectin (CD62p) measurement
Ömer Erdoğan, Mohammed SattarAbduljabbar Al-saadi, Evrim Cevik, Ozge Cevik
doi: 10.14744/ijmb.2021.25238  Pages 192 - 199
INTRODUCTION: Disease-specific biomarkers are an essential tool for the efficient management of pathological conditions, including susceptibility determination, diagnosis, and preventive monitoring. P-selectin (CD62p) is selectively expressed after platelet activation, and is involved in thrombus formation and immune response. The serum and plasma level of CD62p increases in conditions such as heart attack, stroke, some immune diseases, and cancer. The aim of this study was to develop a new, aptamer-based method for CD62p measurement.
METHODS: Aptamers can be used to target specific biomarkers based on their molecular shape. The systematic evolution of ligands by exponential enrichment (SELEX) process is a method to identify aptamers with a high affinity for a specific macromolecular target. This study explored using aptamers to measure CD62p. First, aptamers that specifically bind to CD62p were isolated using the SELEX method. The aptamers that demonstrated the highest binding affinity to the CD62p protein were used to coat 96-well plates. Next, the level of CD62p in human serum was measured using this aptamer and the test performance parameters of sensitivity, specificity, and precision were evaluated.
RESULTS: Among the aptamers used, Apt-1, Apt-2, and Apt-3, bound to CD62p protein with high affinity. Apt-2 had the greatest binding affinity to CD62, demonstrating a binding constant of -9.6 kcal/mol, and a dissociation constant (Kd) of 18.15±2.36 nM. Bovine serum albumin was used in the specificity test as a negative control. No binding between the selected aptamers and this protein was observed. The performance showed that of intra-assay coefficient of variation (CV) was <6.52% and inter-assay CV was <3.96%. The recovery values were between 95.06% and 107.95%, and the linearity values were between 99.49% and 113.17%. Sensitivity was calculated at 0.30 ng/mL of CD62p.
DISCUSSION AND CONCLUSION: The aptamer method to measure CD62p proved to be a sensitive, specific, time-saving, and low-cost option.

10.Flow cytometric evaluation of cancer stem cell markers in HepG2 cells following sorafenib treatment
Yaprak Dönmez Çakıl, Mustafa Erinç Sitar, Zeynep Güneş Özünal, Damla Kayalı, Ranan Gülhan Aktas
doi: 10.14744/ijmb.2021.63935  Pages 200 - 204
INTRODUCTION: Liver cancer is a leading cause of mortality. Sorafenib resistance and cancer stem cells (CSCs) are among the factors that contribute to a poor prognosis. Different drugs enrich different CSC populations with a variety of CSC markers. This study investigated the expression of CSC markers in HepG2 cells in response to low doses of sorafenib using flow cytometry.
METHODS: The cytotoxicity of sorafenib was determined using a cell counting kit-8 assay. The expressions of the CSC markers CD44, CD90, and CD133 were measured with flow cytometry after treatment with sorafenib for 72 hours.
RESULTS: Sofranib inhibited cell proliferation in a dose-dependent manner. Low-dose sorafenib treatment increased CD44 expression; however, there was a decrease in the expression of CD133. An increasing trend was also seen in CD90 expression, but the difference was not significant.
DISCUSSION AND CONCLUSION: The results indicate that CSC expression varied according to the sorafenib dose administered, which supports the role of CSCs as novel pharmacological targets and highlights the importance of their characterization and the ability to identify them.

11.Severe methemoglobinemia caused by prilocaine: A rare case report
Alpaslan Karabulut, Mustafa Şahin
doi: 10.14744/ijmb.2021.17136  Pages 205 - 207
Hemoglobin is a molecule found in erythrocytes that transports oxygen to tissues. Methemoglobin, a form of hemoglobin that can no longer bind oxygen, is formed when the ferrous iron in normal hemoglobin becomes ferric iron as a result of oxidation due to various kinds of oxidative stress. Methemoglobinemia can be congenital, however, exposure to toxins is more often the cause. Local anesthetics can occasionally cause methemoglobinemia, and the potential effects increase with combined or excessive use. Prilocaine-induced methemoglobinemia has been reported, though it is rare. Presently described is the case of an adult case of methemoglobinemia, which developed following a prilocaine injection for fibromyalgia pain. Methylene blue treatment and adjuvant therapy resulted in recovery. This case report serves as a reminder that that prilocaine can be a cause of adult-acquired methemoglobinemia. Methylene blue treatment and adjuvant therapy consisting of infusions of 100% oxygen and 0.9% sodium chloride administered promptly can prevent potentially severe toxic effects.

12.Erroneously high troponin measurement caused by fibrin clot: Two cases
Hatice Bozkurt Yavuz
doi: 10.14744/ijmb.2021.64872  Pages 208 - 210
Acute myocardial infarction is the most common cause of morbidity and mortality in the world. Cardiac troponin measurements play a key role in the diagnosis. However, preanalytical errors as a result of the presence of fibrin or interference due to conditions such as heterophile antibody positivity may cause erroneously high results. Such errors may result in invasive procedures, such as angiography, which may add unnecessary risk. In our hospital, high-sensitivity troponin-I (hs-TnI) was routinely analyzed using a serum separator tube (reference value: female <15.5 pg/mL, male <34.2 pg/mL). This report describes the cases of 2 patients with a false initial troponin measurement: a 19-year-old male patient and a 55-year-old female patient. The hs-TnI value of the male patient was initially measured as 55.5 pg/mL. After the analysis, it was noted that the sample contained fibrin. The sample was centrifuged again and the TnI result was 1.8 pg/mL. Similarly, the TnI result of the female patient was first measured as 90.2 pg/mL. When it was observed that there was fibrin present, the sample was recentrifuged. The revised result was 2.4 pg/mL. The laboratory staff were trained on preanalytical errors, and the use of lithium heparin tubes was implemented in the laboratory as an additional means to eliminate the problem of fibrin interference.

13.Impact of the presence of debris on semen analysis using an automated system (SQA-V)
Claudio Ilardo, Violaine Ostengo, Leatitia Eon, Julie Trentin, Gilles Regnier Vigouroux, Guillaune Quere, Pierre Sanguinet
doi: 10.14744/ijmb.2021.97658  Pages 211 - 215
INTRODUCTION: This study examined the influence of the manual assessment of debris on the results of sperm concentration, progressive motility, and normal morphology measured using the SQA-V system (Medical Electronic Systems LLC, Los Angeles, CA, USA).
METHODS: Sixty samples were analyzed simultaneously and independently by different operators using the manual technique and the SQA-V analyzer. Three measurements (sperm concentration, progressive motility, and normal morphology) of each sample were assessed using the 4 debris assessment levels: none/few, moderate, many, and gross.
RESULTS: Given an optimal assessment of debris, the study data indicate that the SQA-V provided results that had very good agreement with those of the manual method: sperm concentration (rho=0.987, regression analysis formula: y=0.961x+1.962, p<0.0001), progressive motility (rho=0.949, regression analysis formula: y=0.989x+0.418, p<0.0001), and normal morphology (rho=0.694, regression analysis formula: y=0.678x+4.061, p<0.0001). Underestimation of debris increased sperm concentration and decreased motility and normal morphology, while overestimation of debris decreased sperm concentration and increased motility and normal morphology.
DISCUSSION AND CONCLUSION: The results indicated that the performance of the SQA-V remains subject to the competency of the operator.

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