Band 47, Heft 5

Mass spectrometric methods, which transition from chemistry laboratories to clinical laboratories in the basic field, are frequently used in areas such as metabolic analysis, endocrinology, drug research, protein studies and toxicology. Although it is preferred more than immunoassay methods in large-volume laboratories, in certain analyses, it is thought to be an important keystone in clinical laboratory practice. It has found more space in the laboratory area, with users increasing their device experience and working on the applications of problematic tests. It has the potential to take part in metabolic mapping and multiplex analysis, especially in proteomics, with its performance.

Objectives This study aimed to verify the enzymatic ethanol analysis method and compare the technique with headspace gas chromatography (HS/GC). Methods The limit of the blank, limit of detection, and limit of quantification, precision, linearity, and accuracy of the enzymatic process were evaluated based on the data obtained. In addition, a method comparison was made between the enzymatic method and the HS/GC analysis method. Results It was seen that the enzymatic method was linear in the range of 14–450 mg/dL, the coefficient of variation and bias values ranged between 0.5–1.09% and 4.52–9.78%, respectively. The quantitative accuracy was 2.13% and 2.23% at concentrations of 41.16 and 95.82 mg/dL, respectively. When the regression analysis was performed between the results obtained using the enzymatic alcohol dehydrogenase (ADH) and HS/GC methods, y=0.966x + 2.084 linear relationship formula and regression coefficient of r 2 =0.9987, it was found that it showed a good agreement at a confidence interval of 5%. Conclusions The findings showed that the enzymatic method had good linearity, excellent precision, and a high correlation with the higher-order comparator method, the HS/GC method, in the ethanol analysis.

Objectives Diabetes is a chronic group of metabolic disorders those generally present with hyperglycemia hence insulin synthesis defects due to multifactorial causes in beta cells in the Langerhans islets of the pancreas. In the development of diabetes, genetic predisposition is as important as environmental factors. As a result of polymorphism studies in diabetic patients, many genes were associated with the development of diabetes. In our study, we aimed to represent the relationship between diabetes and certain variants of the ghrelin (GHRL) , fat mass and obesity-associated protein (FTO) and peroxisome proliferator-activated receptor-gamma coactivator (PGC-1α) genes which are generally associated with diabetes and obesity. Methods One-hundred type 2 diabetes mellitus (T2DM) patients and ninety-four healthy volunteers were enrolled in our study. GHRL (rs4684677), FTO (rs8044769) and PGC-1α (rs8192678) gene polymorphism studies were performed by the real-time PCR method. Results The carriers of the TT genotype for the FTO (rs8044769) and the GG genotype for the PGC-1α (rs8192678) variants were found more frequently in the patient group, while the GHRL (rs4684677) did not differ between the groups. For the PGC-1α (rs8192678) variant in the patient group, glucose and BMI levels were observed significantly higher in carriers of the GA genotype than those with the GG genotype. There was no statistical difference in the distribution of GHRL (rs4684677) alleles among the groups. Conclusions We conclude that the FTO (rs8044769) and PGC-1α (rs8192678) variants are associated with T2DM in the Turkish population. However, there is no association between GHRL (rs4684677) and T2DM.

Objectives Coronary artery disease (CAD) is a pathological condition resulting from atherosclerosis in the coronary arteries. IL17A has been shown to recruit and activate macrophages in atherosclerotic lesions, thereby participating in plaque destabilization. Currently, whether OLR1 and IL17A variants are involved in the pathogenesis of CAD is unclear. This case-control study aimed to investigate their roles in CAD etiology and prognosis. Methods In this study, 100 severe CAD patients who had undergone the coronary artery bypass graft surgery and 100 healthy controls were genotyped for OLR1 rs11053646, IL17A rs3819025, and rs8193037 variants via RT-PCR. Results The patients with OLR1 rs11053646 CG + GG genotype demonstrated a higher frequency of multi-vessel stenosis (18%) than single- (11.10%) or double-vessel (13.30%) stenosis (p=0.77). Additionally, although not statistically significant, this group of patients had 6.280 times more CAD risk than CC genotype carriers (p=0.089). Furthermore, logistic regression analysis revealed significant associations between the three variants and the risk factors for CAD development, namely waist circumference (p=0.002), body mass index (p=0.013), fasting glucose level (p=0.006), and triglyceride levels (p=0.035). Conclusions OLR1 rs11053646, IL17A rs3819025, and rs8193037 variants do not increase the risk for CAD development. However, this conclusion should be confirmed with a larger cohort.

Objectives Sequence variants in cytokine genes are related to affect cytokine gene levels. In this study, it was aimed to examine eight single nucleotide polymorphisms (SNPs) in five cytokine genes (TNF-α, INF-γ, IL-6, IL10, TGF-β) for the Turkish population living in Marmara region and to reveal the genetic distance between the study group and other populations. Methods In this study, three-hundred unrelated healthy individuals were involved and all genotyping were performed by using sequence-specific primers PCR (PCR-SSP) method. The SNP data were analyzed for Hardy Weinberg equilibrium fit by calculating expected genotype frequencies and comparing them to the observed values using Arlequin software version 3.1. The genetic distances between the study group and other populations were calculated and a neighbor-joining tree was constructed by PHYLIP. Results The observed genotypes of TNF-α (−308), IFN-γ (+874), TGF-β (codon 10), and TGF-β (codon 25) of the subjects were found to be similar with other populations investigated in this study. However, there is a significant frequency difference for IL-6 and IL-10 genotypes between populations. Conclusions The current population study provided more reference values for these polymorphisms and generated a control group to be used in further association studies especially for transplantation, GVHD, autoimmune and malign disease.

Objectives To find BRCA1/2 test selection criteria unique to the Turkish population, as well as to provide the BRCA1/2 gene mutation distributions of patient population to the literature. Methods Genetic counseling was given to 2,373 cases with a family history of high-risk breast and/or ovarian cancer who applied to Istanbul University, Oncology Institute, Department of Cancer Genetics between 1994 and 2021 and selected by NCCN Guidelines for the BRCA1/2 test criteria. In our clinic, mutation screenings in BRCA1/2 genes were performed by Sanger sequencing method in patients admitted between 1994 and 2014 and by NGS method in patients admitted between 2015 and 2021. Results The overall mutation rate in our patient group selected from high-risk patients was 16.5% (391/2,373) after BRCA1/2 gene mutation screening performed in 2,373 cases who applied to the Cancer Genetics clinic. Of the patients with mutations, 57.5% (225/391) had BRCA1 mutation, 41.9% (164/391) had BRCA2 mutation, and 0.6% (2/391) had both BRCA1 and BRCA2 pathogenic mutations. People diagnosed before the age of 60 who have a history of triple-negative breast cancer had a 28.5% overall mutation rate. Conclusions BRCA1/2 mutation in Turkish population were evaluated in accordance with NCCN BRCA1/2 genetic test selection criteria; we discovered that all of our study results were statistically significant (p<0.05).

Objectives Periodontitis, a dental disease that causes inflammation of gums is triggered by a bacterial infection. Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1) has been reported to participate in inflammatory diseases. In the present study, we investigated the effects of lncRNA MALAT1 on periodontal ligament cells (PDLCs) with lipopolysaccharide (LPS) induction. Methods MALAT1, microRNA-155 (miR-155), Sirtuin 1 (SIRT1), Bax, and Bcl-2 RNA expressions were detected by using real-time quantitative polymerase chain reaction (RT-qPCR). PDLC viability and apoptosis were assessed by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium salt (MTT) assay and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) assessed secretions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Luciferase reporter test was applied for confirming the binding of miR-155 with MALAT1 and SIRT1, respectively. Results Overexpression of MALAT1 facilitated the LPS-induced periodontitis while MALAT1 suppression restrained LPS-induced injury. Also, miR-155 was expressed in LPS-induced PDLCs. Moreover, MALAT1 negatively regulated miR-155 followed by the up-regulation of SIRT1 which in turn reduced the inflammation in LPS-induced PDLCs. Conclusions We concluded that upregulated lncRNA MALAT1 could accelerate periodontitis through the regulation of miR-155/SIRT1. Our findings suggested a novel MALAT1/miR-155/SIRT1 pathway for the treatment of periodontitis.

Objectives Gene therapy using adenoviruses has shown tremendous promise in animal and human models in the past. The tumor responses achieved by these viruses were either because of their oncolytic properties or therapeutic genes expressed from the genome of the virus. We have previously shown enhanced viral replication in cell models that have lost or lower expression of the cyclin dependent kinases inhibitor (p21/Waf-1). Beside the early onset of many viral genes in the p21/Waf-1 knock out cell model, we observed a significantly higher copy number for the viral VA1 but non VA2 non-coding RNA transcripts. In this study we investigated the effect of adenoviral VA1 non-coding RNA on the landscape of cellular proteins. Methods Using a DU145 cell line as a model that was transiently transfected with a plasmid carrying the adenoviral VA1 non-coding RNA, we were able to study changes in the proteome and the cellular cascade of the cells. Using state of the art global proteomics analyses of the differentially expressed proteins between the VA1 overexpressed and control cells demonstrated how the early onset of VA1 transcript affected cellular machinery. Results Using a 1.5 fold cut off between the down-regulated or overexpressing proteins, we were able to demonstrate how the non-coding VA1 gene might be responsible for suppressing the proteasome degradation, the endocytic and lysosomal pathways. Similarly, overexpression of the non-coding VA1 transcript was responsible for the upregulation (1.5 fold) of approximately 40 different proteins in the spliceosome machinery. Conclusions These data demonstrated how the non-coding VA1 RNA functioned independently of any other viral proteins in modulating the cell signaling cascades to promote viral propagation. To our knowledge this is the first report to investigate the function of VA1 non-coding RNA on the cellular proteome.

Objectives This study aimed to evaluate the effects of pomegranate fruit extract (PFE) on levels of transient receptor potential (TRP) channel and caspase-3 (Casp-3) expressions, tumor necrosis factor-α (TNF-α), total sialic acid (TSA), reduced glutathione (GSH), and malondialdehyde (MDA) in mice with induced colorectal cancer (CRC) by investigating effects of PFE on in vitro mitotic index (MI). Methods Different PFE concentrations on​​ MI against 0.3 μg/mL mitomycin-C (MMC) in cell culture were evaluated by binocular light microscopy. During in vivo applications on Balb/c mice, it was given once physiological saline to group I, PFE for ten weeks to group II, a single dose of azoxymethane (AOM) plus dextran sulfate sodium in drinking water (DSS) to group III, and AOM plus DSS plus PFE to group IV. Tissue samples were evaluated by western blotting, spectrophotometric, and histopathological methods. Results Expressions of Casp-3, TRP ankyrin 1 (TRPA1), and melastatin 2 (TRPM2) channels and TNF-α, TSA, GSH, and MDA concentrations in evaluated tissues had significantly better levels in PFE-treated groups compared to CRC-induced mice. Conclusions Results of the present study indicate that PFE application in mice with induced CRC may be an important modulator of TRPA1 and TRPM2 channels, apoptosis, and inflammatory response by decreasing oxidative stress.

Objectives A small subpopulation of colorectal cancer stem cells (CSCs) possess the ability to self-renew and the capacity to initiate the original tumor. EpCAM high /CD44 + cells are regarded as CSCs in colorectal cancer. The present study was undertaken to investigate the significance of EpCAM in the in vitro proliferation ability and oxaliplatin chemoresistance of EpCAM high /CD44 + colorectal CSCs. Methods We applied fluorescence-activated cell sorting (FACS) to separate the EpCAM high /CD44 + subset from human colorectal cancer cell line HCT116. We also used siRNA targeting EpCAM to create EpCAM − /CD44 + subpopulation. Then we compared EpCAM high /CD44 + cells and EpCAM − /CD44 + cells for proliferation ability and the chemoresistance to oxaliplatin by CCK8 assay. Results The EpCAM high /CD44 + subset comprises almost 6.25 ± 0.09% in cell line HCT116, and the EpCAM − /CD44 + cells displayed a significantly lower proliferation ability and weaker oxaliplatin chemoresistance than the EpCAM high /CD44 + cells. Conclusions EpCAM is critical for tumor proliferation and oxaliplatin chemoresistance in EpCAM high /CD44 + colorectal CSCs.

Objectives Thrombotic thrombocytopenic purpura (TTP) is a rare thrombotic microangiopathy with no standardized prognostic model to predict mortality. The aim of the study is to determine parameters associated with TTP-related mortality. Methods In this cross-sectional, retrospective study, 77 TTP patients, treated with therapeutic plasma exchange between 2001 and 2019 in Ankara University Faculty of Medicine were included. Results There was no significant relationship between ADAMTS13 inhibitor levels, activity and mortality. Median number of plasmapheresis was 10 (2–32), higher in patients with complete response. Anemia, kidney injury and LDH levels were associated with survival; there was no significant relationship between platelet counts at the time of diagnosis and mortality. Mortality was lower in patients with platelet counts above 100 × 10 9 /L and normalized LDH after treatment. Hemoglobin, albumin, LDH and creatinine levels at the time platelet counts exceeded 50 × 10 9 /L were associated with survival. Conclusions We determined several clinical and laboratory parameters associated with mortality. Fewer numbers of plasmapheresis was associated with mortality; thus other treatments as rituximab and caplacizumab should be considered early in non-responders. Including changes in laboratory parameters may be considered in prognostic scoring systems to be developed in the future.

Objectives Mesenteric ischemia is a surgical emergency caused by poor blood supply to the intestines. In ischemia, the decrease in blood flow to the tissue causes acidosis and cell death through anaerobic metabolism. Clinoptilolite is one of the most abundant natural zeolites, and it is used for its ion exchange and adsorbent properties. Clinoptilolite has been reported to have an immune-enhancing, anti-carcinogenic, and antioxidant effect in-vitro / in-vivo studies. Clinoptilolite’s histological and biochemical effects on ischemic small intestines. Methods The experimental animals were randomly divided into sham, control, and clinoptilolite treatment group. Clinoptilolite was administered intraperitoneally after ischemia/reperfusion. Cardiac blood was stored for biochemical analysis. Total antioxidant levels and total oxidant levels were analyzed from the sera taken from groups. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) expressions in blood samples were determined by RT-qPCR. At the end of the reperfusion, terminal ileum tissues were taken for histological tests. Results The mean TNF-α expression level was 3.89 in the control group and 2.91 in the clinoptilolite treatment group. The mean IL-6 expression levels were 2.32 in the control group and 1.49 in the clinoptilolite treatment group. Conclusions clinoptilolite administration provided healing in the rat ischemia-reperfusion injury model.

Objectives The main etiological factor in intrauterine growth restriction (IUGR) is the impairment of the fetoplacental unit. Due to the placental endothelial disintegrity and vascular permeability disruptions, endocan has been an interesting molecule to search for associations with IUGR. The aim of this study was to investigate the umbilical cord blood endocan concentrations in IUGR pregnancies. Methods This cross-sectional case-control study was conducted on 50 IUGR-complicated (patients) and 50 control pregnancies. The demographics and clinical findings were recruited from records. The umbilical cord blood was studied for endocan concentrations. Results The patient group compared to controls had significantly more previous IUGR history, lower maternal weight gain, an earlier gestational week at delivery, and more cesarean sections (p<0.001). The umbilical cord endocan concentrations were significantly higher in patients than in controls (p<0.001). Oligohydramnios (n=38) was significantly higher in the patients and had a significant relationship with high endocan concentrations (p<0.001). Moderate and mild positive correlations were found among endocan concentration and biparietal diameter/head circumference and femur length values, respectively, while a moderate negative correlation was observed in abdominal circumference values (p<0.001). Conclusions A significant increase in the umbilical cord blood endocan concentrations was found in IUGR pregnancies.

Objectives This study was conducted to evaluate whether sildenafil effectively treats necrotizing enterocolitis (NEC). Methods Thirty-eight rat pups were divided into 4 groups: control, sildenafil-control, NEC, and sildenafil-NEC (Sil-NEC). NEC was induced by hypoxia/reoxygenation and cold stress. The pups were treated by administering 1 mg/kg sildenafil by intraperitoneal injection once a day until the fourth postnatal day. The tissues were stained with hematoxylin/eosin staining and examined with the TUNEL test for apoptosis. The intestinal levels of malondialdehyde (MDA), interleukin 1β (IL-1β), inducible nitric oxide synthase (iNOS), caspase-3, and glutathione peroxidase (GSH-px) activity were quantified. Results TUNEL positivity (p=0.002) and intestinal damage grade (p<0.001) were found to be significantly lower in the Sil-NEC group. In addition, MDA, IL-1β, iNOS, caspase-3 levels, and GSH-px activity were also found to be significantly lower in the Sil-NEC group (p<0.001, p=0.004, p=0.011, p=0.026, p=0.002 respectively). Conclusions In this study, sildenafil has been shown to reduce intestinal damage and prevent the development of necrosis biochemically and histopathologically, with its antioxidant, anti-apoptotic, and anti-inflammatory effects, in the treatment of the experimental necrotizing enterocolitis model. This may suggest that sildenafil can be used to treat necrotizing enterocolitis, but further clinical studies are required.

Objectives The aim is to investigate the usefulness of lactate dehydrogenase (LDH)/Albumin, LDH/Lymphocyte and LDH/Platelet ratios on the prognosis of coronavirus disease (COVID-19) Alpha (B.1.1.7) variant pneumonia. Methods A total of 113 patients who were diagnosed with COVID-19 pneumonia and 60 healthy control group were included in this study. The cases were divided into 2 as classic COVID-19 group, and COVID-19 B.1.1.7 variant group. Complete blood count (CBC) and biochemical parameters of the patients were analyzed retrospectively. Patients with COVID-19 B.1.1.7 variant group were also grouped according to the length of stay in the hospital and the days of hospitalization. Results LDH/Albumin, LDH/Platelet, and LDH/Lymphocyte ratios were found to be higher in COVID-19 B.1.1.7 variant group when compared to the control group (p<0.001). The ferritin, neutrophils/lymphocyte (NLR) ratio, procalcitonin (PCT) and LDH/Albumin had the highest area under the curve (AUC) values in the COVID-19 B.1.1.7 variant group (0.950, 0.802, 0.759, and 0.742, respectively). Albumin, Lymphocytes and hemoglobin values were significantly higher in the COVID-19 B.1.1.7 variant group than in the classic COVID-19 group (p<0.05). Conclusions LDH/Albumin and LDH/Lymphocyte ratios may be useful for clinicians in predicting the risk of progression to pneumonia in COVID-19 B.1.1.7 variant patients.

Objectives Low vitamin D levels are associated with the severity and mortality of COVID-19 infection. Nevertheless, the relationship between the 25-hydroxyvitamin D [25(OH)D] levels and the antibody response following COVID-19 vaccination is not fully elucidated. Herein, we explored the relationship between SARS-CoV-2 IgG (sCOVG) and 25(OH)D. Methods In this prospective observational case-control study, we used an automated chemiluminescent immunoassay method to measure sCOVG and 25(OH)D levels in 96 patients 28 days following the second dose of inactivated vaccine. We considered the positivity for sCOVG at three different index values: 1, 2.42, and 7. We classified 25(OH)D levels between 0 and 20 ng/mL as vitamin D deficiency, 21–29 ng/mL as insufficiency and 30 ng/mL as sufficiency. Results Median sCOVG index was 6.02 (interquartile ranges 3.41–11.63) and median 25(OH)D level was 11.5 ng/mL (interquartile ranges 10–17). We could not find a significant correlation between 25(OH)D and sCOVG levels (Spearman’s rho, r=0.175, p=0.12). When considering the variables categorically, we did also not conclude significant relationships between adequate or inadequate antibody responses in patients with deficient, insufficient, and sufficient 25(OH)D by three sCOVG cut-off index values (1, 2.42, and 7) (Chi-square test, p=0.8, 0.29, and 0.08, respectively). Conclusions The relevant literature is limited on the association between the antibody response to COVID-19 vaccines and vitamin D levels. Although the previous research suggested conflicting findings of the response to mRNA vaccines, we could not conclude a significant relationship between sCOVG and 25(OH)D levels 28 days after two doses of inactivated COVID-19 vaccine.

Objectives Studies have shown that fibrinolysis activity is insufficient in COVID-19 patients. Plasminogen activator inhibitor-1 (PAI-1) is an important antifibrinolytic molecule that plays a key role in the fibrinolytic system. In our study; we aimed to evaluate serum PAI-1 and other biochemical parameters of COVID-19 patients in terms of disease course and mortality. Methods A total of 40 COVID-19 patients were hospitalized in the service and intensive care unit (ICU) of our hospital from October to December 2020 and 20 healthy volunteers were included in our study. The patients were grouped as those who transferred to the ICU from the service and transferred to service from the ICU. The first and second values of the same patients in both the service and the ICU were analyzed by SPSS. Results The PAI-1 levels of the patients in the ICU were significantly higher than the levels of the same patients in the service and the healthy control group (p<0.001). IL-6, ferritin, and D-dimer levels in the ICU of the same patients were significantly higher than the levels of service and healthy control group (p<0.001). A positive correlation was found between initial serum PAI-1 and D-dimer levels in patients hospitalized in the service (p=0.039) and initial serum ferritin and IL-6 levels in the ICU (p=0.031). Conclusions In our study, we found that PAI-1 levels increased significantly with the increase in mortality in COVID-19 patients.

Objectives For a definitive diagnosis of COVID-19, respiratory tract samples are evaluated by polymerase chain reaction (PCR). In our study, PCR using a tear sample was used to diagnose COVID-19, and it was questioned whether it was a screening method. Unlike the general practice, Schirmer strips were used instead of a swab for tear sample collection in this study. In addition, the diagnostic values of serum procalcitonin (PCT), C-reactive protein (CRP), and Neutrophil (NEU) count in predicting COVID-19 disease from tears were also questioned. Methods A total of 94 patients who were positive for COVID-19 by PCR test were included in this study. Tear samples were obtained from patients with Schirmer strips, commonly used in eye examination, and studied with the PCR technique. CRP, PCT value, and NEU count were also compared between the positive and negative groups of the PCR. The obtained data were analyzed using the R Studio software, and the results were considered statistically significant for p<0.05. Results Of these patients, 61 (64.9%) tear PCR was negative, and 33 (35.1%) tear PCR was positive. The mean age was 61.72 ± 17.62 years. The patients were divided into two groups: tear PCR positive and negative. There was no significant age difference between these groups. As a result of ROC Analysis; When serum PCT, CRP, and NEU % values were examined in predicting COVID-19 disease from tears, it was seen that CRP (p=0.027) and especially PCT (p=0.003) values of patients with PCR-positive were significantly higher. Conclusions PCR study on tears collected with Schirmer strips is a different and non-invasive method, but it was concluded that the proposed method could not be used as a screening test. In addition, significantly higher serum PCT values were found in patients with COVID-19 positivity in tears (p<0.05).

Objectives Heterophile antibodies can cause interferences in immunometric assays. While many tests are shown to be affected by interference from heterophilic antibodies, the D-dimer test has rarely been reported to be affected. With this case, we report an elevated D-dimer measurement which was not compatible with the clinical presentation. Case presentation A 41-year-old patient who was admitted to hospital with heart palpitations had a D-dimer elevation irrelevant to his clinical condition. D-dimer measurements were repeated in new samples directly and after being treated in heterophilic blocking tube with two different reagent lots of a latex-based automated immunoturbidimetric assay and an immunoturbidometric assay. D-dimer values were normalized (0–0.5 mg/L) when we used a new lot of reagent on the same instrument or measured by an immunoturbidometric method on the chemistry analyzer. After treatment with HBT, all samples revealed D-dimer results within the reference ranges. Conclusions The presence of heterophile antibodies in a sample should be considered when an elevated D-dimer value that is not compatible with the clinical presentation is encountered. Apart from the patient’s HA’s causing false results, sporadic susceptibility of the reagents should also be kept in mind as a possibility.