Band 50, Heft 3

The preanalytical phase is the basis upon which the accuracy and reliability of laboratory testing mostly depend. Despite many recent advancements in laboratory medicine, preanalytical errors remain the most common source of diagnostic inaccuracies, accounting for up to 70 % of total laboratory mistakes. These errors can significantly impact patient outcomes, enhance healthcare costs, and impair laboratory efficiency. This article is hence aimed at exploring the various complexities of the preanalytical phase, examining the clinical and economic consequences associated with preanalytical errors, especially focusing on blood drawing and spurious hemolysis, which represent the most frequent causes of preanalytical problems in clinical laboratories.

Objectives Diabetic retinopathy (DR) is a common complication of diabetes mellitus (DM), with a risk of blindness. The study was to search for effective biomarkers of DR. Methods The key molecules of DR were mined by bioinformatics. Dual luciferase reporter gene assay, knockdown and overexpression assay were used to detect the regulatory relationship between hsa_circ_0053004, hsa-miR-646, and CBX2 , and their clinical value was evaluated by constructing ROC curve. Levels of hsa_circ_0053004, hsa-miR-646, and CBX2 mRNA was detected by RT-qPCR, and western blotting was for detection of CBX2 protein expression. CCK-8 reagent and transwell assay were used for cell viability and migration detection. Results Hsa_circ_0053004 and CBX2 expressed highly in DR patients, while hsa-miR-646 was defective. Hsa_circ_0053004 down-regulated hsa-miR-646 by targeting it directly. Hsa-miR-646 inhibited CBX2 expression by targeting its 3′UTR. Overexpression of hsa_circ_0053004 or CBX2 promoted the high glucose-induced enhancement in human retinal microvascular endothelial cell (HRMEC) viability and migration, which was reversed by co-transfection of hsa-miR-646 mimic. The AUC of hsa_circ_0053004, hsa-miR-646, and CBX2 in predicting DR was 0.827, 0.805, and 0.817, respectively, and their combined AUC was 0.942. Conclusions Hsa_circ_0053004 promoted CBX2 expression by adsorbing hsa-miR-646, thereby promoting HRMECs to grow and migrate, and they may be good biomarkers of DR.

Objectives Breast cancer (BC) is a malignant tumor characterized by high heterogeneity. The current study aims to examine the process underlying the LINC00578/miR-495-3p/RNF8 regulatory pathway in BC progression, aiming to discover new therapeutic targets. Methods The levels of LINC00578, miR-495-3p, and RNF8 were quantified. The prognostic significance of LINC00578 was assessed through the utilization of Kaplan-Meier survival curves along with Cox regression analysis. The proliferation, migration, and invasion abilities of BC cells were assessed. The targeting relationships between LINC00578 and miR-495-3p, along with between miR-495-3p and RNF8, were verified using a dual luciferase reporter assay. Results LINC00578 and RNF8 were significantly elevated in BC tissues and cells, while miR-495-3p was lowly expressed. Compared to BC patients with high expression, those with low expression of LINC00578 exhibit substantially higher 5-year overall survival rate. LINC00578 expression, Lymph Node Metastasis (LNM), and tumour, node, and metastasis (TNM) stage were independent prognostic indicators. LINC00578 targeted and regulated miR-495-3p, and knockdown of miR-495-3p overturned the inhibitory action of LINC00578 interference on the proliferation, invasion, and migration of BC cells. Furthermore, miR-495-3p targeted and regulated RNF8, and knocking down RNF8 reversed the stimulatory effects of miR-495-3p interference on RNF8 expression. Conclusions The upregulation of LINC00578 is associated with the deterioration of BC and indicates a poor prognosis. LINC00578 expression, LNM, and TNM stage were independent prognostic indicators. LINC00578 influences the advancement of BC by targeting and regulating the miR-495-3p/RNF8 axis.

Objectives Polycystic ovarian syndrome (PCOS) is a heterogeneous endocrine condition that influences 15–20 % of women at their fertile age. In this academic work, we aimed to research serum anti-mullerian hormone (AMH) levels using the Vidas® and Atellica® methods and other hormone parameters in PCOS. Methods This study included 55 controls and 55 PCOS women diagnosed by Rotterdam criteria. AMH levels were measured by Vidas® and the Atellica® devices, and the other parameters were obtained from Ankara Bilkent City Hospital’s Laboratory Information System. Results The AMH levels utilized by the Vidas® (p<0.001) and the Atellica® (p=0.001) were both more elevated in PCOS women. The Vidas® and the Atellica® methods were well correlated, and the correlation coefficient value was 0.899. The AMH cut-off value was 3.23 μg/L with a sensitivity of 65.5 % and a specificity of 34.5 % by utilizing the Vidas® method and 3.66 μg/L with a sensitivity of 65.4 % and a specificity of 36.4 % by utilizing the Atellica® method. PCOS women’s serum AMH levels were positively correlated with fasting glucose and luteinizing hormone (LH) and negatively correlated with age and progesterone. The PCOS group’s levels of serum estradiol (E2) were notably more elevated and levels of serum prolactin were notably lower than the controls. Conclusions This study showed that serum AMH levels measured with both the Atellica® and the Vidas® present significant differences between PCOS patients and the controls but still, AMH alone is not diagnostic enough to distinguish PCOS patients from controls.

Objectives We aimed to compare the anti-angiotensin II type I receptor (AT1R) antibody levels considering the development of anti-HLA antibodies after transplantation, pathologically confirmed humoral or vascular rejection reactions and late complications. Methods Forty-nine hypertensive patients who had presented to the Division of Nephrology, Transplantation Outpatient Clinic of Memorial Health Group Hospitals Transplant Units and Istanbul Medical Faculty for live kidney transplantation and underwent transplantation between 2015 and 2019, were included in our study. Among our patients (29 males/20 females), 76 % had primary hypertension, and 24 % had secondary hypertension. In this study, serum AT1R antibody levels before and after transplantation were measured using the Enzyme-Linked ImmunoSorbent Assay (ELISA) method. The anti-HLA antibody was detected with the Luminex method using the single antigen bead (SAB) assay. Results The AT1R antibody was found positive in 7 (18.9 %) of 37 patients who had primary hypertension and in 4 (33.3 %) of 12 patients who had secondary hypertension (p=0.426). The examination of the frequencies of development of post-transplant anti-HLA antibodies showed that the frequency of AT1R antibody positivity was found higher in the group with positive anti-HLA antibodies compared to the levels in the group with negative anti-HLA antibodies (p=0.002). The comparison of the pre-transplant and post-transplant values showed that the AT1R antibody level was statistically significantly decreased after transplantation [7.3/6.6 (3.2–21.1) vs. 6.2/4.7 (1.5–24.5), p=0.001]. Conclusions The findings of our study suggest that the evaluation of the AT1R antibody levels among non-HLA antibodies may be important for predicting the immunological risk before transplantation.

Objectives The onset and advancement of silicosis are intricately linked to immune system activation and oxidative stress. We aimed to examine whether occupational silica exposure induces alterations in neopterin levels, the immune system, or the oxidant/antioxidant system. Methods Fifty-two healthy individuals, 47 workers with silica exposure who did not develop the disease, and 71 silicosis patients were included in the study, totaling 170 participants. Neopterin was measured in serum and urine. Chromatographic analysis of neopterin level was performed with Thermo Ultimate 3000 ultra-high-performance liquid chromatography system. Tumor necrosis factor alpha (TNF-α), interferon-gamma (IFN-γ), fibroblast growth factor (FGF), interleukin (IL)-1α, IL-1β, 8-hydroxy-2-deoxyguanosine (8-OH-dG), glutathione reductase (GR), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were measured in serum. Immune and oxidative stress parameters were measured by spectrophotometric and spectrofluorometric methods. Results Serum neopterin levels were significantly different between the study groups, while urinary neopterin levels were not. Serum neopterin levels were significantly decreased in the silicosis group compared to the exposure and non-exposed group groups. TNF-α, IFN-γ, FGF, IL-1α, 8-OH-dG, and GR increased with silica exposure, while SOD and GPx decreased. Serum neopterin, IL-1α, GR, age, and forced expired volume (FEV1%) were found to be risk factors for silicosis development. A negative correlation existed between neopterin levels, TNF-α, and overall exposure duration. Conclusions Neopterin is applicable in the diagnosis and monitoring of oxidative stress and alterations in the immune system associated with silicosis. Thus, it can ensure that preventive measures are taken at an early stage.

Objectives Autism Spectrum Disorder (ASD) is a neurological and developmental disorder that affects all aspects of social communication, with stereotypical and limited interest and atypical responses to sensory stimuli. There isn’t a reliable lab test that can help clinicians diagnose ASD. This study aimed to examine serum lipocalin-1, resistin, cathepsin-D, neurokinin A, agmatinase, brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and glial fibrillary acidic protein (GFAP) levels in children with ASD. Methods The study had 40 matched control subjects and 39 ASD sufferers in total. The diagnosis was made by medical professionals using the Gilliam Autistic Disorder Rating Scale-2-TV (GOBDÖ-2-TV), Denver Developmental Screening Test, and Porteus Intelligence Tests. Results Compared to the control group, serum levels of lipocalin-1, resistin, cathepsin-D, agmatinase, BDNF, and NGF were found to be high and GFAP levels were low in children having ASD. B12 and magnesium levels were significantly lower in the ASD group than in the control group, while sodium and chloride levels were higher in the ASD group (p<0.05). Conclusions The aforementioned parameters may be involved in the pathophysiology of ASD, as evidenced by the fact that their values were greater in children with ASD than in controls. Additionally, evaluation of some routinely measured biochemical parameters may suggest that nutritional and electrolyte balance may differ in individuals with ASD.

Objectives This study emphasizes the importance of determining the serum levels of pentraxin-3 (PTX3), fibroblast growth factor-2 (FGF2), and tumor necrosis factor-stimulated gene-6 (TNFAIP6) in patients with Crimean-Congo hemorrhagic fever (CCHF). Methods This prospective study involved 30 confirmed CCHF patients and 30 healthy controls. Serum concentrations of PTX3, FGF2, and TNFAIP6 were quantified utilizing a quantitative sandwich ELISA method. Results CCHF patients exhibited markedly elevated PTX3 levels, reflecting an acute inflammatory response. As a long pentraxin, PTX3 functions as a pattern recognition receptor that activates the complement system to aid in pathogen clearance. Additionally, FGF2 levels were significantly increased, indicating a potential role in repairing endothelial damage. Known for promoting angiogenesis and immune regulation, FGF2 may counteract endothelial dysfunction induced by CCHF. Conversely, TNFAIP6 levels were lower in patients, possibly due to shifts in cytokine activity that suppress its anti-inflammatory and extracellular matrix-regulating effects, potentially leading to greater tissue injury. Conclusions The dysregulation of PTX3, FGF2, and TNFAIP6 in CCHF patients signifies a disrupted equilibrium in inflammatory and vascular response mechanisms. This triad of biomarkers could serve as a valuable tool for assessing the severity of CCHF and may present therapeutic targets for modulating inflammation and mitigating endothelial damage. Achieving a balance among PTX3, FGF2, and TNFAIP6 could be instrumental in alleviating disease complications, thereby suggesting a potential therapeutic strategy for managing CCHF effectively.

Objectives The Pasteur effect in cellular energy metabolism is defined as the switch to anaerobic glycolysis due to alterations in ambient oxygen concentration or inhibition of the electron transport chain (ETC) and oxidative phosphorylation. This study aimed to determine the effect of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection on the energy metabolism of leukocytes among Coronavirus Disease 2019 (COVID-19) patients at various stages. Methods A total of 90 cases were analyzed in asymptomatic, mild, moderate, and severe COVID-19 patients. Levels of aerobic glycolysis intermediates and products of anaerobic glycolysis were measured in leukocytes with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results A decrease in the rate of aerobic glycolysis was found as the disease progressed (p-values<0.001), while the anaerobic glycolysis rate was unchanged in all study groups because lactate levels had no difference between asymptomatic, mild, moderate, and severe groups (p=0.754). Moreover, the ratios of lactate to Tricarboxylic Acid (TCA) cycle parameters demonstrated a significant reduction in the rate of aerobic glycolysis of COVID-19 patients at different stages (p-values<0.001). Furthermore, Receiver Operating Characteristic (ROC) curve analysis for citrate, alpha-ketoglutarate, fumarate, lactate, lactate/citrate, lactate/alpha-ketoglutarate, and lactate/fumarate had significant cut-off values in Moderate and Severe patients (p<0.001). Conclusions Our results suggest that the Pasteur effect from aerobic to anaerobic glycolysis was shown to be induced as the severity of COVID-19 progresses.

Objectives Sepsis is an organism’s exaggerated response causing physiological and pathological biochemical changes. This study aims to investigate the alterations in thiol/disulfide ratio (TD) and ischemia-modified albumin (IMA), as well as their association with acute phase reactants in sepsis patients. Methods The study was conducted in the Anesthesia and Resuscitation Intensive Care Unit between October 1, 2023, and October 30, 2024, and included 66 patients, 29 women and 37 men. Patients were classified according to their Sequential Organ Failure Assessment (SOFA) scores. The parameters measured in this study included C-reactive protein (CRP), procalcitonin (PCT), native thiol, total thiol, disulfide, disulfide/native thiol × 100, disulfide/total thiol × 100, native thiol/total thiol × 100 ratios and IMA. Results In the SOFA<10 group, acute phase reactants such as CRP and PCT were elevated on day 0; however, these levels decreased significantly on days 3 and 7. In the SOFA≥10 group, all measured values decreased on day 7, although the changes were not statistically significant (p>0.05). IMA values were significantly higher on day 0 (p<0.05). In both groups, patients had elevated levels of total thiols, native thiols, and IMA, suggesting increased antioxidant levels. Conclusions Our study’s findings indicate that IMA and TD are involved in the pathogenesis of sepsis in patients.

Objectives Oxidative stress occurs when free radicals accumulate at high levels in the body, affecting biological molecules. Some studies have focused on the multisystemic involvement, signs of premature aging, and the potential role of oxidative stress in the pathogenesis of myotonic dystrophy type 1 (DM1). We aimed to evaluate the levels of oxidative stress markers in patients diagnosed with DM1. Methods In a cross-sectional study, 27 clinically and genetically confirmed DM1 patients and 34 healthy individuals matched for age and gender were included. Oxidative stress markers, including ischemia-modified albumin (IMA), total antioxidant status (TAS), total oxidant status (TOS) levels, ischemia-modified albumin to albumin ratio (IMAR), and oxidative stress index (OSI) were examined. Results TOS (p=0.001), IMA levels (p=0.001), IMAR (p=0.004), and OSI (p<0.001) in the patient group were significantly higher than in the healthy group, while TAS (p<0.001) was lower in the patient group. Conclusions The elevation of oxidative stress markers suggests the potential effect on the pathogenesis of DM1. Thus, antioxidant therapy approaches could be crucial to reducing oxidative stress in patients with DM1.

Objectives Microbial fermentation has long been known as a method for processing teas to raise products with novel biological activities. Our study aimed to chemically characterize flavonoid components in fungal mediated biotransformed Camellia chrysantha (Hu) Tuyama leaves. Methods The fermented leaves of Camellia chrysantha were prepared by inoculation with an Eurotium cristatum fungal consortium and extracted with ethanol combining ultrasound-assisted technique, before introducing to chemical characterization. Normal and reverse phase C18 column chromatography and thin layer chromatography were applied for fractionation and isolation of compounds from the ethanolic extract. Nuclear magnetic resonance spectra ( 1 H NMR, 13 C NMR, HSQC and HMBC) were recorded for structure determination, while in vitro bioassays of free radical scavenging capacity and α -glucosidase inhibition were employed to assess anti-oxidative and anti-diabetic properties of isolated compounds. Results Four flavonoids, i.e. quercitrin ( 1 ), afzelin ( 2 ), (-)-epicatechin ( 3 ), and engelitin ( 4 ) were isolated from fermented Camellia chrysantha (Hu) Tuyama leaves. Compounds 1 and 3 were found to exhibit scavenging property against both DPPH and ABTS. Compound 2 was noticeably the most effective against α -glucosidase, presenting an IC 50 of 78.25 μg/mL. Conclusions The investigation on chemical composition of microbial fermented leaves of Camellia chrysantha (Hu) Tuyama brought out flavonoid compounds with significant biological activities, which could serve as a basis for further possible application in manufacturing health protecting natural products.

Objectives The present study identifies the level changes of markers (Apelin, Elabela, Meteorin, ALT, AST, GGT) in the liver after acute intake of high-dose paracetamol and their role in determining liver damage. Methods The rats were categorized into two groups (Control and Toxication), each consisting of seven rats. Apelin, Elabela, Meteorin, ALT, AST and GGT levels were determined in liver tissue. The levels of Apelin, Elabela, and Meteorin in liver tissue homogenate were assessed following the kit procedure, utilizing commercial enzyme-linked immunosorbent assay. ALT, AST and GGT analyzes were performed from liver tissue homogenate with Erba colorimetric kit according to the kit procedure. It was also evaluated histopathologically from liver tissue. Results Looking at the findings, a significant decrease was determined in the Elabela and Meteorin data in the Apap group compared to the control group. In the Apelin data, a statistical increase was observed in the Apap group compared to the control group. A significant increase in ALT analysis was detected in the APAP group compared to the control group. Considering GGT and AST data, a statistical increase was detected in the APAP group compared to the control group.While Bax, Bcl-2 and iNOS immunoreactivity was not observed in the control group, a yellowish-brown positive reaction was detected in the cytoplasm of degenerated and necrotic hepatocytes, especially around the vena centralis, in the paracetamol group. Conclusions As a result, it is thought that Apelin, Elabela and Meteorin markers may be decisive markers in determining toxicity in liver damage.

Objectives This research aimed to examine the anticancer properties of cosmosiin, a natural flavonoid, on human lung carcinoma cells by in silico molecular docking, network pharmacology, and in vitro experiments. Methods The targets of cosmosiin and targets related to lung cancer were retrieved from various databases. The common targets between cosmosiin and lung cancer were identified by venny online server followed by construction of the protein-protein interaction (PPI) network. Further, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment were performed to identify main genes involved along with the signalling pathways affected. The hub genes were used for in silico molecular docking to identify molecular interaction between these targets and cosmosiin. In vitro experiments which consisted of MTT cell viability, clonogenic, cell apoptosis and cell migration assays validated the network pharmacology results. Results Network pharmacology identified 63 common targets between cosmosiin and lung cancer and out of these 63, eight protein targets were found to be most important based on their involvement in numerous signalling pathways in lung cancer. Two (NFKB1 and PIK3R1) out of eight targets showed highest degree values and were subjected to in silico molecular docking which showed cosmosiin showing strong binding the active sites of these two target proteins with PIK3R1 showing higher binding energy value (−9.7 kcal/ml) than NFKB1 (−7.5 kcal/mol). GO and KEGG enrichment analysis revealed key gene functions, molecular functions, cellular components as well as key signalling pathways involved in the treatment of lung cancer by cosmosiin. MTT and apoptotic assays indicated that cosmosiin induced concentration-dependent cytotoxic and apoptotic induction effects in A-549 human lung cancer cells respectively. Cell migration assay exhibited that cosmosiin treatment at varying doses led to a concentration-dependent suppression of cell migration hinting towards the anti-metastatic potency of cosmosiin against lung carcinoma. Conclusions In conclusion, the present study provides strong theoretical and experimental evidence of the anticancer, apoptotic and anti-metastatic potential of cosmosiin natural product against lung carcinoma along with the detailed mechanism of action involving various biological targets, cellular components and signalling pathways.

Objectives After reviewing the reference product specifications, found that they include the SEC HPLC result but do not specify Protein A chromatography as a test. Although there is no record of reference product specifications, alternative methods are necessary, particularly for routine analysis of biotechnological products. Methods The SEC mobile phase consisted of 0.1 M sodium phosphate and 0.2 M saline buffer pH 6.80 run isocratically at a flow rate of 0.5 mL/min. The Protein A chromatography column was equilibrated (60 mM sodium phosphate buffer, pH 7.4 (binding buffer), 60 mM sodium phosphate buffer, pH 2.5 (elution buffer) gradient system) at 1 mL/min. Results The SEC HPLC method validation study was conducted, ranging from (0.01–2.0 mg/mL) 1.0 % (LLOQ level) to 200 %. The Protein A chromatography method between (0.0078 and 4.0 mg/mL) 0.78 % (LLOQ level) and 400 % linearity levels. In the linearity study, both ANOVA and Graphpad results were shown. Both bioanalytical methods were performed by establishing validation parameters for the mAb reference product in accordance with the ICH M10 guideline. Conclusions Additionally, the findings shed light on routine titer analysis of biotechnological products, as there is no documentation in the literature regarding Protein A chromatography validation.

Objectives Atellica UAS800 (Siemens Healthineers, Germany), an automatic microscopic analysis device, based on the principle of analyzing microscopic images with a digital camera, has the feature of discriminating pathogens such as cocci and rod apart from the complete urinalysis. We aimed to evaluate the usability of the Atellica UAS800 microscopic analyzer as a screening tool for identifying urinary pathogens by comparing it with the gold-standard urine culture method in our study. Methods A total of 1,056 urine samples between June 2022 and July 2022 in Ankara City Hospital were included. Atellica UAS800 was used for complete urinalysis. Simultaneous samples obtained for urine culture were processed in Walk Away Specimen Processor (WASP; Copan, Italy). After evaluating WASP, cultures with growth were identified by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight, Mass Spectrometry (VITEK-MS; bioMerieux Diagnostics, France) device at the species level. Results Of the 1,056 urine samples, 551 (52.2 %) and 505 (47.8 %) were from male and female patients respectively. The ROC curve analysis of the 1,056 samples yielded AUC values of 0.743 and 0.865 for the detection of bacteria and rods, respectively. In the ROC curve analysis regarding gender, the AUC was found to be 0.772 and 0.867 in female patients and 0.693 and 0.859 in male patients in the detection of bacteria and rod respectively. Conclusions The Atellica UAS800 automatic microscopic analyzer has a high specificity of detecting gram-negative bacilli, which are responsible for approximately 50–90 % of UTIs and can guide clinicians in the empirical treatment of UTIs.

Objectives B vitamins are a group of eight essential nutrients involved in various biological processes, including energy production and the proper functioning of the nervous system. They play a critical role in certain inherited metabolic diseases (IMDs). Deficiencies in the metabolism of thiamine (B1), biotin (B7), and cobalamin (B12) can lead to serious health issues, including confusion, seizures, respiratory problems, encephalopathy, coma, hearing and vision loss, ataxia, and more. The main objective of our study is to highlight the role of Gas Chromatography-Mass Spectrometry (GC/MS) in detecting vitamin B-responsive IMDs in Moroccan patients. Case presentation Urine samples from symptomatic individuals suspected of having intermediate metabolism deficiencies, excluding aminoacidopathies, were analyzed using GC/MS. Three cases of IMDs were identified: pyruvate dehydrogenase complex deficiency, biotinidase deficiency, and methylmalonic aciduria. Each patient presented with neurological symptoms and metabolic abnormalities. Following targeted vitamin supplementation (B1, B7, or B12), significant clinical improvement was observed, with resolution of neurological manifestations and normalization of brain CT findings. Conclusions Urinary metabolite screening using GC/MS in patients with neurological and other symptoms indicative of IMDs can facilitate practical, reliable diagnosis and accessible management. GC/MS analysis is a highly sensitive, rapid, and precise technique for quantifying a wide range of metabolites in human urine. We recommend considering such analysis as a routine test for specific symptoms in the context of personalized and functional medicine practice.