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Study design:Experimental study.Objectives: To determine the neuroprotective effects of zinc and melatonin on spinal cord ischemia-reperfusion (I/R) injuries of rabbits.Setting: The Experimental Research Centre of Selçuk University, Konya, Turkey.Methods:Twenty-four male rabbits underwent spinal cord ischemia by clamping the thoraco-abdominal aorta for 20 min. Twenty minutes before the aortic clamping, animals received zinc, melatonin or a combination of both. Neurological examination of the animals was performed three times during reperfusion period. The animals were killed 24 h after reperfusion. Spinal cord samples were taken for biochemical and histopathological evaluation.Results:Pre-treated animals with zinc, melatonin or combination displayed better neurological outcomes than the I/R group (P<0.05). Zinc, melatonin and combined treatment prevented spinal cord injury by reducing apoptosis rate (P<0.05) and preserving intact ganglion cell numbers (P<0.05). Zinc pre-treatment protected spinal cord by preventing malondialdehyde (MDA) formation (P=0.002), increasing glutathione peroxidase (GPx) activity (P=0.002) and decreasing xanthine oxidase enzyme activity (P=0.026) at molecular level. Melatonin treatment also resulted with MDA formation (P=0.002), increased GPx activity (P=0.002) and decreased xanthine oxidase activity (P=0.026).Conclusion:The results of the study showed that prophylactic zinc and melatonin use in spinal cord I/R not only suppressed lipid peroxidation by activating antioxidant systems but also had significant neuroprotective effects by specifically improving the neurological and histopathological situation.Spinal Cord (2007) 45, 722-730; doi:10.1038/sj.sc.3102035; published online 13 February 2007. [ABSTRACT FROM AUTHOR]

Tube manufacturers use different composition of gels and blood clot activator formulations in serum tube production. Our aim was to investigate the within-tube (repeatability) and between-tube variation, concordance between comparison results of BD and VacuSEL tubes. Blood samples were collected from control subjects (n = 20) and patients (n = 30) in accordance with the CLSI GP41-A6 and CLSI GP34-A guidelines. Twenty-three clinical chemistry parameters were analysed via Roche Cobas C702 Chemistry Analyzer on T0 (0 hour) and T24 (24 hour). Mean differences % were compared with Wilcoxon matched pair test. Clinical significance was evaluated based on desirable bias according to total allowable error (TEa). VacuSEL tubes demonstrated acceptable performance for the results of 20 parameters with regards to desirable bias % limits. Lactate dehydrogenase (LD) [mean difference % (%95 confidence intervals (CI) values of BD and VacuSEL tubes at T0 [6.41% (4.80-8.01%)]; sodium (Na) and total protein (TP) at T24 [-0.27% (-0.46 to -0.07%) and -1.39% (-1.87 to -0.91), respectively] were over the desirable bias limits (LD: 4.3%, Na: 0.23% and TP: 1.36%, respectively) but not exceeding total biological variation CV % [Na: 0.5 (0.0-1.0) % and TP: 2.6 (2.3-2.7) %). %95 confidence intervals (CI) of T0 LD values overlap with within-subject biological variation % (CI) limits (LD: 5.2 (4.9-5.4) %). The differences between two tubes were not medically significant and necessarily conclusive. VacuSEL serum tubes presented comparable performance with BD serum tubes.

Introduction: Our study aimed to compare the short- and particularly long-term type 2 diabetes mellitus (T2DM) remission prediction abilities of ABCD, individualised metabolic surgery (IMS), DiaRem2, Ad-DiaRem and DiaBetter scoring systems in Turkish adult type 2 diabetic morbidly obese patients who underwent bariatric surgery., Patients and Methods: Our study was planned as a retrospective cohort study. A total of 137 patients with T2DM, including 78 sleeve gastrectomy (SG) and 59 Roux-en-Y gastric bypass (RYGB) patients, were included in the 1st-year evaluation after bariatric surgery, and a total of 115 patients with T2DM, including 64 SG and 51 RYGB patients, were included in the evaluation at the end of the 5th year., Results: In the 1st year after bariatric surgery, area under the ROC curve (AUC) values for diabetes remission scores were 0.863 for Ad-DiaRem, 0.896 for DiaBetter, 0.840 for DiaRem2, 0.727 for ABCD and 0.836 for IMS. At 5 years after bariatric surgery, the AUC values for diabetes remission were 0.834 for Ad-DiaRem, 0.888 for DiaBetter, 0.794 for DiaRem2, 0.730 for ABCD and 0.878 for IMS., Conclusions: According to our study, the DiaBetter score provided a better AUC value than the other scores both in the short and long term but showed similar predictive performance to Ad-DiaRem in the short term and IMS in the long term. We believe that DiaBetter and Ad-DiaRem scores might be more appropriate for short-term assessment and DiaBetter and IMS scores for long-term remission assessment., (Copyright © 2024 Copyright: © 2024 Journal of Minimal Access Surgery.)

Objectives: Urinalysis is a first-line test for screening for urinary tract infection. Several devices performing strip and sediment analysis have been introduced. The aim of this study was to compare the performance of Labsan Tricell-1000 and Dirui FUS-2000 automated urine analyzers with manual microscopy., Methods: 463 urine samples were analyzed. Digital image processing and particle recognition automatically display the cells in a flowing sheath fluid mixed monolayer urine sample, take the pictures of particles via digital camera, analyse these pics with a particle recognition software, transfer images of the formed elements to the screen and allow well-trained personnel to select, reclassify or remove them. Manual microscopy was used for comparison., Results: Agreement between Tricell-100 and manual microscopy was very good for RBC (ϰ = 0.80), and WBC (ϰ = 0.83); good for CaOx (ϰ = 0.69), SEC (ϰ = 0.80), YLC (ϰ = 0.72), HC (0.69) and LC (ϰ = 0.64); moderate for BAC (ϰ = 0.51), APC (ϰ = 0.43) and MT (ϰ = 0.55); fair for GC (ϰ = 0.39) and RTEC (ϰ = 0.32)., Conclusions: Labsan Trion TriCell-1000 demonstrated satisfactory performance and can be used in routine urinalysis. In the case of low counts of RBC, presence of yeast, crystal, casts or cell clumping in urine sediment, characterization of urine particles should be performed by manual microscopy., Competing Interests: None., (© 2024 The Authors.)

Objectives: Data generation in clinical settings is ongoing and perpetually increasing. Artificial intelligence (AI) software may help detect data-related errors or facilitate process management. The aim of the present study was to test the extent to which the frequently encountered pre-analytical, analytical, and postanalytical errors in clinical laboratories, and likely clinical diagnoses can be detected through the use of a chatbot., Methods: A total of 20 case scenarios, 20 multiple-choice, and 20 direct questions related to errors observed in pre-analytical, analytical, and postanalytical processes were developed in English. Difficulty assessment was performed for the 60 questions. Responses by 4 chatbots to the questions were scored in a blinded manner by 3 independent laboratory experts for accuracy, usefulness, and completeness., Results: According to Chi-squared test, accuracy score of ChatGPT-3.5 (54.4 %) was significantly lower than CopyAI (86.7 %) (p=0.0269) and ChatGPT v4.0. (88.9 %) (p=0.0168), respectively in cases. In direct questions, there was no significant difference between ChatGPT-3.5 (67.8 %) and WriteSonic (69.4 %), ChatGPT v4.0. (78.9 %) and CopyAI (73.9 %) (p=0.914, p=0.433 and p=0.675, respectively) accuracy scores. CopyAI (90.6 %) presented significantly better performance compared to ChatGPT-3.5 (62.2 %) (p=0.036) in multiple choice questions., Conclusions: These applications presented considerable performance to find out the cases and reply to questions. In the future, the use of AI applications is likely to increase in clinical settings if trained and validated by technical and medical experts within a structural framework., (© 2023 Walter de Gruyter GmbH, Berlin/Boston.)

Background: Tryptophan is an essential amino acid primarily metabolized by the kynurenine pathway in mammals. Intermediate metabolites emerging in this pathway have been associated with many neurogenerative diseases. This study aimed to compare tryptophan pathway metabolite levels in patients with multiple sclerosis (MS) and healthy controls and reveal the relationship of tryptophan metabolites with disease subtype and the Expanded Disability Status Scale (EDSS) score., Methods: The study included a total of 80 MS cases [53 with relapsing remitting MS (RRMS) and 27 with secondary progressive MS (SPMS)] and 41 healthy volunteers. The patients with RRMS were further divided into relapse (RRMS-attack) and non-attack (RRMS-stable) groups. Using liquid chromatography mass spectrometry, tryptophan, kynurenine, kynurenic acid, quinolinic acid, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid levels were measured. The serum metabolite levels of the patient and control groups were compared. In addition, the link and relationship between the EDSS score and disease duration of the patients and their plasma tryptophan metabolite levels were examined., Results: The tryptophan level of the patient group was significantly lower than that of the healthy controls (p<0.05). The kynurenine (105.38±65.43), quinolinic acid (10.42±3.56), kynurenine/tryptophan ratio (0.0218±0.019), and quinolinic acid/kynurenic acid ratio (1.7054±0.96141) of the patients with MS were significantly higher compared to the controls (p<0.05). In the receiver operating characteristic analysis of the power of kynurenine/tryptophan and quinolinic acid/kynurenic acid ratios in predicting the disease, both ratios predicted the diagnosis of MS (area under the curve: 0.793 and 0.645, respectively; p<0.05), albeit at low sensitivity and specificity. The parameters were similar between the RRMS-attack and RRMS-stable patient groups (p>0.05). There was also no significant difference between the RRMS and SPMS patient groups in terms of tryptophan metabolites (p>0.05). Lastly, no significant relationship was observed between tryptophan metabolites and MS subtype and the EDSS score., Conclusion: Our findings revealed that the kynurenine pathway involved in the tryptophan metabolism differed between the patients with MS and healthy controls, and this difference may be a limited guide in the diagnosis of MS, due to major overlaps in values for MS versus Controls, and is insufficient to determine the disease subtype., Competing Interests: Declaration of Competing Interest There is no conflicts of interest in this article., (Copyright © 2023. Published by Elsevier B.V.)

Objectives: Recent studies have shown that derangements in kynurenine pathway metabolite levels are associated with various pathologies such as neurodegenerative diseases, schizophrenia, depression, bipolar disorder, rheumatoid arthritis, and cancer. Therefore, reliable, accurate, fast, and multiplex measurement methods for kynurenines have become increasingly important. This study aimed to validate a new mass spectrometric method for analyzing tryptophan metabolites., Methods: A tandem mass spectrometric method, including protein precipitation and evaporation steps, was developed to measure serum levels of tryptophan, kynurenine, kynurenic acid, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid. Samples were separated using a Phenomenex Luna C18 reversed-phase column. The kynurenine pathway metabolites were detected by tandem mass spectrometry. The developed method was validated according to Clinical & Laboratory Standards Institute (CLSI) guidelines and applied to hemodialysis samples., Results: The developed method was linear at the concentrations of 48.8 - 25,000, 0.98 - 500, 1.2-5000, 1.2-5000, and 0.98-250 ng/mL for tryptophan, kynurenic acid, kynurenine, 3-hydroxyanthranilic acid, and 3-hydroxykynurenine, respectively. The imprecisions were less than 12 %. The median serum concentrations of tryptophan, kynurenine, kynurenic acid, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid were 10530, 1100, 218, 17.6, and 25.4 ng/mL in pre-dialysis blood samples, respectively. They were 4560, 664, 135, 7.4, and 12.8 ng/mL in post-dialysis blood samples, respectively., Conclusions: A fast, simple, cost-effective, accurate, robust, and validated tandem mass spectrometric method was developed, and the method was successfully used for the quantitation of kynurenine pathway metabolite concentrations in hemodialysis patients., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 THE AUTHORS.)

Objectives: Primary Sjögren's syndrome (pSS) is an autoinflammatory disease characterized by inflammation of the exocrine glands. Elevated inflammation causes an increase in kynurenine pathway (KP) metabolite levels by activating indoleamine 2,3-dioxygenase (IDO). The aim of this study was to measure serum KP metabolite concentrations in patients with pSS and to evaluate the relationship between these metabolites with disease activity score and clinical manifestations., Design & Methods: A total of 80 patients with pSS and 80 healthy controls were enrolled in this study. Serum tryptophan (TRP), kynurenine (KYN), kynurenic acid (KYNA), 3-hydroxyanthranilic acid (3HAA), 3-hydroxykynurenine (3HK), quinolinic acid (QUIN) concentrations were quantified with liquid chromatography with tandem mass spectrometry (LC-MS/MS). Demographic characteristics, clinical manifestations and disease activity score (ESSDAI) of the participants were recorded., Results: The serum level of KYN and QUIN were significantly higher in patients with pSS with low and moderate activity compared those healthy controls, while the serum level of TRP, KYNA/KYN and 3HK/KYN were lower. In addition, the significant difference for the serum level of KYNA was only in patients with moderate activity from healthy controls, and the difference was higher in favor of pSS patients. Moreover, the KYN/TRP levels were significantly increased with disease activity. The ESSDAI score was positively correlated with KYN/TRP ratio, but negatively correlated with KYNA/KYN ratio., Conclusions: These findings indicated that KP metabolites may play a role in the etiopathogenesis, activation and progression of pSS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Background: This study evaluated the potential of Wharton's jelly mesenchymal stem cells with high tolerogenic properties in reducing immunosuppressive dosage and related adverse effects., Methods: A 4- to 6-week-old, 30-40 g weight, male inbred CD57BL/6 mice were used as skin allograft donors, whereas Balb/c mice with similar characteristics were used as recipients. Wharton's jelly stem cells were obtained from a commercial kit sourced from human umbilical cord. Skin allografts were performed from CD57Bl6 to Balb/c mice (day 0). Group 1 (control) received no treatment. Group 2 received 15 mg/kg cyclosporin A on days 0 to 30. Group 3 received 5.7 × 10 6 and 10.3 × 10 6 cell/kg Wharton's jelly stem cells on days 0 and 3, respectively. Groups 4, 5, and 6 received a combination of 15, 10, and 5 mg/kg per day cyclosporine A (days 0 to 30) with the same stem cell dose with group 3, respectively. Graft rejection was evaluated with digital photography and thermal imaging, histopathology (Banff grading, epithelialization scores, dermoepidermal dissociation), immunochemistry (Ki-67 and Bcl-2), and biochemical methods (interleukin 10, interleukin 2, interferon γ, tumor necrosis factor α) (day 10). Cumulative adverse effects of cyclosporin A occurring in the groups were revealed by histopathological evaluation of kidney and liver (a modified semiquantitative method of infiltration of inflammatory cells around the portal area and lobular region in liver; modification of the Banff rating of proximal tubules and hypertrophia of juxtaglomerular apparatus cells in kidney) (day 30)., Results: There was no rejection in groups 2, 4, and 5 until the end of study. These were statistically different versus groups 1 (day 10 ± 0.71), 3 (day 11 ± 0.82), and 6 (day 11 ± 0.58) (all P 's < 0.05). Groups 4 and 5 have exhibited statistically similar findings in histopathological (4 epithelization score: 3.7 ± 1.3; 5 epithelization score: 3.5 ± 0.5; 4 Banff grading score: 0.8 ± 0.6; 5 Banff grading score: 1.0 ± 0.5; both P 's = 1.00), immunohistochemical (4 Bcl-2 score: 3.5 ± 0.5, P = 0.618; 5 Bcl-2 score: 3.4 ± 0.5, P = 1.00; 4 Ki-67 score: 3.7 ± 0.4, P = 1.00; 5 Ki-67 score: 3.5 ± 0.5, both P 's = 1.00), and levels of cytokines (both P 's = 1.00) versus group 2. Adverse effects on kidneys and liver were lowest and statistically similar in groups 3, 5, and 6 (all P 's = 00) versus group 1., Conclusions: Wharton's jelly mesenchymal stem cells alter bioavailability of cyclosporine, albeit at much lower doses and with fewer systemic adverse effects., Competing Interests: Conflicts of interest and sources of funding: The authors state that they have no conflicts of interest to disclose and no funding was received to assist in the creation of this manuscript., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Introduction: This study aims to compare the predictive capacity of ABCD, DiaRem2, Ad-DiaRem, and DiaBetter scoring systems for type 2 diabetes mellitus (T2DM) remission in Turkish adult morbidly obese patients who underwent SG. Methods: This retrospective cohort study included 80 patients who underwent sleeve gastrectomy (SG) operation who were diagnosed with T2DM preoperatively, and had at least one-year follow-up after surgery. Because bariatric surgery is performed on patients with class III obesity (BMI ≥ 40 kg/m2) or class II obesity (BMI ≥ 35 kg/m2) with obesity releated comorbid conditions in our hospital, our study cohort consisted of these patients. Results: The diagnostic performance of the DiaBetter, DiaRem2, Ad-DiaRem and ABCD for identifying diabetes remission, assessed by the AUC was 0.882 (95% CI, 0.807-0.958, p < 0.001), 0.862 (95% CI, 0.779-0.945, p < 0.001), 0.849 (95% CI, 0.766-0.932, p < 0.001) and 0.726 (95% CI, 0.601-0.851, p = 0.002), respectively. The AUCs of the Ad-Diarem, DiaBetter and DiaRem2 were statistically higher than AUC of the ABCD (all p-value < 0.001). Besides, there was no statistically significant difference in AUCs of the Ad-Diarem, DiaBetter and DiaRem scores (all p-value > 0.05). Conclusion: Ad-Dairem, DiaBetter and DaiRem scoring systems were found to provide a successful prediction for diabetes remission in sleeve gastrectomy patients. It was observed that the predictive power of the ABCD scoring system was lower than the other systems. We think that the use of scoring systems for diabetes remission, which have a simple use, will become widespread., (© 2022 Suleyman Baldane et al., published by Sciendo.)

Objective. This study was aimed to evaluate the prevalence of Cushing's syndrome and the diagnostic performance of the 1 mg dexamethasone suppression test in class 3 obese patients. Methods. Anthropometric measurements and other laboratory data, including 1 mg dexamethasone suppression test of 753 class 3 obese patients, who applied to the Endocrinology and Metabolism Outpatient Clinic for the pre-bariatric surgery evaluation between 2011 and 2020, were evaluated retrospectively. Results. An abnormal response to the 1 mg dexamethasone suppression test (cortisol ≥1.8 mcg/dl) was observed in 24 patients and the presence of Cushing's syndrome was confirmed by additional tests in 6 patients. The prevalence of abnormal dexamethasone suppression test was 3.18% and the prevalence of Cushing's syndrome 0.79%. The specificity value was determined as 97.5% for 1 mg dexamethasone suppression test with cortisol threshold value ≥1.8 mcg/dl. Conclusions. The prevalence of Cushing's syndrome was found to be low in class 3 obese patients and 1 mg of dexamethasone suppression test had a very sufficient performance for Cushing's syndrome screening in this patient group., (© 2022 Suleyman Baldane et al., published by Sciendo.)

Aim: The aim of this study was to measure serum levels of methylarginine derivatives and related metabolites in patients with gout. Materials & methods: This study enrolled 100 patients with gout and 80 patients in the control group. Serum asymmetric dimethylarginine, symmetric dimethylarginine, L-N-monomethylarginine, arginine, homoarginine, citrulline and ornithine levels were measured with tandem mass spectrometry. Results: Serum ornithine, citrulline and total methylated arginine load levels were statistically significantly higher in patients with gout compared with the control group, while serum arginine and homoarginine levels and global arginine bioavailability ratio were statistically significantly lower. Conclusion: There may be an association between gout, methylarginine levels and hyperuricemia and increased risk of cardiovascular disease.

Our aim in this study was to develop a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the measurement of atorvastatin, rosuvastatin and their major metabolites, and furthermore to evaluate patients' adherence to statin therapy and to investigate the effect of statin therapy on various hematological and biochemical parameters. A simple protein precipitation was performed for the extraction of analytes and the extracted samples were injected directly. The levels of drugs and their metabolites were measured by the validated method in a total of 210 patients diagnosed with unstable angina pectoris (USAP), ST-elevation myocardial infarction (STEMI) or non-ST-elevation myocardial infarction (NSTEMI). Various biochemical and hematological parameters were measured. The linearity ranges for atorvastatin and rosuvastatin were 1.22-2,500 and 0.97-2000 ng/ml, respectively. The inter-assay coefficient of variation for all analytes was ≤ 6%. In patients diagnosed with USAP, STEMI and NSTEMI, treatment compliance rates were 22.1, 23.5 and 41.2% for atorvastatin and 36.1, 40.2 and 67.1% for rosuvastatin, respectively. An economical, simple and reliable measurement method has been developed. Our findings support the poor patient compliance with statin therapy in the included population. It was observed that 6 months of statin treatment caused slight changes in biochemical and hematological parameters., (© 2022 John Wiley & Sons Ltd.)

Behçet disease (BD) is an inflammatory, multisystemic vasculitis of unknown etiopathogenesis. However, innate and adaptive immune system involvement and immune-mediated networks play a vital role in the inflammatory cascade. Indoleamine 2,3-dioxygenase 1 (IDO1) is activated in chronic inflammatory states and catalyzes the first and rate-limiting step of tryptophan (TRP) metabolism along the kynurenine pathway (KP). The study aimed to measure KP metabolites levels in patients with BD and investigate the relationship between disease activity and clinical findings with these metabolites. The study included 120 patients with BD and 120 healthy volunteers. Serum TRP, kynurenine (KYN), kynurenic acid (KYNA), 3-hydroxyanthranilic acid (3HAA), 3-hydroxykynurenine (3HK), and quinolinic acid (QUIN) levels were measured with the tandem mass spectrometric method. Demographic data, clinical manifestations, and disease activity score (BDCAF) were recorded. Serum KYN, KYNA, 3HK, 3HAA, QUIN levels, and KYN/TRP ratio were higher (p < 0.05) in patients with BD compared to the control group, while TRP levels were lower (p < 0.05). KYN/TRP ratio and QUIN levels were significantly higher in the presence of neuro-Behçet, while serum KYN levels were significantly higher in the presence of arthritis (p < 0.05). In addition, serum QUIN levels were significantly higher in the presence of thrombosis (p < 0.05). BDCAF score positively correlated with KYN/TRP ratio. Our findings showed that serum KP metabolite levels were elevated in patients with BD, and there is a relationship between these metabolites with disease activity, clinical findings, and inflammatory burden., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Objective: In this study, the objective was to evaluate the cardiovascular and metabolic effects in men with male pattern alopecia beginning before 30 years of age., Methods: Total of 81 people (41 androgenetic alopecia (AGA) and 40 healthy individuals) were included in the study. Twenty-four-hour ambulatory blood pressure (ABP) measurement, high sensitive C-reactive protein (hsCRP), galectin-3 were studied. Hamilton-Norwood scale (HNS) was used to determine the AGA types of the cases., Results: The mean age in the AGA and control groups was 30.3 ± 7.5 and 30.8 ± 6.0, respectively. Twenty-four-hour ABP measurements, hsCRP, and galectin-3 were similar in both groups. There was a positive correlation between HNS grade with age, BMI, triglyceride levels and fasting blood glucose levels in individuals with AGA. Similarly, there was a positive correlation between HNS grade with daytime pulse wave velocity and night-time reflection magnitude. A significant positive correlation was determined between hsCRP with BMI and waist circumference, and between galectin-3 with BMI, waist circumference, hip circumference, HOMA-IR in individuals with AGA., Conclusions: This study shows that AGA patients are similar to the normal population in terms of insulin resistance or metabolic syndrome components. However, hsCRP and galectin-3 appear to be associated with cardiovascular disease risk factors in individuals with AGA.

Hydroxychloroquine has attracted attention in the treatment of COVID-19. Many conflicting findings have been reported regarding the efficacy and safety of this drug, which has been used safely in the rheumatological diseases for years. However, these studies lacked measurement methods that allow accurate assessment of hydroxychloroquine and its metabolite levels. The aim of this study was to measure hydroxychloroquine and its metabolite levels in whole blood samples of patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), Sjogren's syndrome (SS) and scleroderma (Scl) by a robust, simple and accurate validated tandem mass spectrometric method, and to investigate the relationship between these levels with drug-related adverse effects and disease activity scores. The validated LC-MS/MS method was applied to measure blood hydroxychloroquine and its metabolite levels of patients with RA, SLE, SS, Scl. Various haematological and biochemical parameters were measured with Beckman-Coulter AU 5800 and Beckman Coulter LH 780 analyzers, respectively. QTc intervals were calculated with Bazett's formula, and the patients were followed up by clinicians in terms of clinical findings and adverse effects. Hydroxychloroquine levels of patients were similar to previous studies. There was a negative correlation between disease activity scores and hydroxychloroquine levels, while the highest correlation was between QTc interval, creatinine and GFR levels with desethylchloroquine. Bidetylchloroquine had the highest correlation with RBC count and liver function tests. Our findings showed that hydroxychloroquine and its metabolite levels were associated with disease activity scores, renal, hepatic function, QTc prolongation, and hematological parameters., (© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Objectives: The study aimed to develop a validated LC-MS / MS method for the measurement of carbamazepine and carbamazepine-10,11-epoxide (CBZE) levels, to compare the carbamazepine concentrations measured by AbSciex API 3200 LC-MS/MS and Beckman Coulter Emit® 2000 immunoassay and to investigate the effect of carbamazepine concentrations on various hematological and biochemical parameters., Methods: For the measurement of carbamazepine and CBZE levels, a validated LC-MS / MS method was developed. Serum carbamazepine levels of patients were measured by AbSciex API 3200 LC-MS / MS and Beckman Coulter Emit® 2000 immunoassay. Biochemical, hematological parameters, and hormone levels were measured by Beckman-Coulter AU 5800 (Beckman Coulter, Brea, USA), Beckman Coulter LH 780 (Beckman Coulter, Miami, FL, USA), and Cobas 6000 (Roche Diagnostics, Germany) analyzers, respectively., Results: The imprecision values for all analytes were less than 7.0 %. The correlation coefficient between the methods was 0.981 (95 % confidence interval: 0.975 to 0.985). Linear regression analysis demonstrated highly significant associations of carbamazepine concentrations with albumin (B = -0.300, p = 0.040), calcium (B = -0.262, p = 0.004), phosphorus (B = -0.241, p = 0.022), WBC (B = -0.288, p = <0.001), PLT (B = -0.236, p = 0.003), RBC (B = -0.257, p = 0.001), NEU% (B = -0.289, p = <0.001), LYM% (B = -0.268, p = 0.001), D vitamini (B = -0.344, p = 0.006) levels., Conclusions: A robust, rapid, and simple method has been developed. Our study revealed that carbamazepine and its metabolite have a significant correlation and likely impact on bone metabolism, blood cell counts, serum protein, albumin levels, and electrolyte concentrations., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Various studies reported that increased proinflammatory cytokines in patients with ankylosing spondylitis (AS). Proinflammatory cytokines can affect the expression of various kynurenine pathway enzymes and therefore lead to metabolic changes that can affect the inflammatory response and immunity. Our aim was to measure serum levels of kynurenine pathway metabolites in patients with AS., Methods: The study included 85 patients with AS and 50 healthy volunteers. Serum tryptophan, kynurenine, kynurenic acid, 3-hydroxyanthranilic acid, 3-hydroxykynurenine, quinolinic acid concentrations were measured with tandem mass spectrometry. In addition, participants were divided into four groups according to the treatment regimen: TNF-α inhibitor group, conventional therapy group, control group and newly diagnosed AS group. These groups were compared in terms of kynurenine pathways metabolites, interleukin 6 (IL-6), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels., Results: Serum tryptophan, kynurenic acid, 3-hydroxykynurenine levels were significantly decreased (p < 0.05) in both AS groups compared to the control group, while the levels of kynurenine, quinolinic acid, CRP, ESR, and IL-6 were higher (p < 0.05). The Kynurenine/Tryptophan ratio and CRP levels of the conventional therapy and anti-TNF therapy group were significantly lower than the newly diagnosed AS patients (p < 0.05)., Conclusion: As a result of our study, we found that altered kynurenine pathway metabolism in patients with AS. Conventional therapy and anti-TNF-α therapy are effective in reducing the Kynurenine/Tryptophan ratio and CRP levels, although the effect of both treatments on other metabolites appears to be limited., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Our aim in this study was to measure serum levels of methylarginines and related metabolites in patients with ankylosing spondylitis (AS), moreover, to investigate the relationship between these parameters and various clinical and laboratory parameters of patients with AS. The study included 60 patients with AS and 60 healthy volunteers. Serum asymmetric dimethylarginine (ADMA), L-N monomethylarginine (L-NMMA), symmetric dimethylarginine (SDMA), arginine (Arg), homoarginine (hArg), ornithine, and citrulline concentrations were measured with tandem mass spectrometry. In addition, participants were divided into three groups according to the treatment regimen: TNF-α inhibitor group (n = 25), conventional therapy group (n = 35), and control group (n = 60). These groups were compared in terms of serum levels of methylarginine pathway metabolites and various biochemical parameters. It was found that total methylated arginine load significantly increased in patients with AS (p < 0.001), and the Arg/ADMA ratio was positively correlated with HDL levels and negatively correlated with glucose, ESR, total cholesterol, triglyceride, and LDL levels. In addition, serum ADMA, SDMA, total methylated arginine load, and CRP levels were lower (p < 0.05) in the TNF-α group compared to the conventional treatment group. To the best of our knowledge, this is the first study to comprehensively investigate serum methylarginine levels in patients with AS. Elevated total methylated arginine load and decreased global arginine bioavailability ratio (GABR) indicate that NO metabolism is impaired in patients with AS. Therefore, the increased cardiovascular risk in patients with AS may be related to the decreased NO production or bioavailability due to the elevated total methylarginine load., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Favipiravir is a broad-spectrum inhibitor of viral RNA polymerase. It is currently used as a possible treatment for coronavirus disease 2019 (COVID-19). Pre-clinical or clinical trials of favipiravir require robust, sensitive, and accurate bioanalytical methods for quantitation of favipiravir levels. Recently, several studies have been reported about developing a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring favipiravir levels. However, these methods were validated predominantly for plasma samples, electrospray ionization was operated only in negative or positive mode, and clinical application of these methods has not been applied for patients with COVID-19. This study aimed was to develop a validated LC-MS/MS method for the measurement of favipiravir levels in positive and negative electrospray ionization mode and to perform a pilot study in patients with COVID-19 receiving favipiravir to demonstrate the applicability of this method in biological samples. Simple protein precipitation was used for the extraction of favipiravir from the desired matrix. Favipiravir levels were quantitated using MS / MS with an electrospray ionization source in positive and negative multiple reaction monitoring (MRM) mode. The chromatographic detection was performed on a reverse-phase Phenomenex C18 column (50 mm × 4.6 mm, 5 µm, 100 Å) with gradient elution using 0.1% formic acid in water and 0.1% formic acid in methanol as mobile phase. The method was linear over the concentration ranges of 0.048-50 µg/mL (in negative ionization mode) and 0.062-50 µg/mL (in positive ionization mode) with a correlation coefficient (r 2 ) better than 0.998. The total run time was 3.5 min. The intra-assay and inter-assay %CV values were less than 7.2% and 8.0%, respectively. A simple, rapid and robust LC-MS / MS method was developed for the measurement of favipiravir and validation studies were performed. The validated method was successfully applied for drug level measurement in COVID-19 patients receiving favipiravir., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Multiple sclerosis (MS) is an autoinflammatory, chronic central nervous system disease. In the pathogenesis of the disease increased nitric oxide (NO) levels play an important role. Nitric oxide (NO) has neuroprotective effects in physiological conditions, however, it is thought that excessive NO formation in MS may lead to demyelination and axonal damage. Derivatives of methylarginine including asymmetric dimethyl arginine (ADMA), L-N monomethyl arginine (L-NMMA), symmetric dimethyl arginine (SDMA) directly or indirectly reduce NO production. Our aim was to measure the levels of methylarginine derivatives and citrulline, ornithine, arginine, homoarginine levels, which are metabolites associated with NO metabolism, in MS subgroups., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Nitric oxide (NO) plays an important role in endothelial homeostasis. Asymmetric dimethyl arginine (ADMA), L-N monomethyl arginine (L-NMMA) and symmetric dimethyl arginine (SDMA), which are derivatives of methylarginine, directly or indirectly reduce NO production. Therefore, these metabolites are an important risk factor for various diseases, including cardiovascular diseases. Numerous methods have been developed for the measurement of methylarginine derivatives, but various difficulties have been encountered. This study aimed to develop a reliable, fast and cost-effective method for the analysis and measurement of methylarginine derivatives (ADMA, SDMA, L-NMMA) and related metabolites (arginine, citrulline, homoarginine, ornithine), and to validate this method according to Clinical and Laboratory Standards Institute (CLSI) protocols., Methods: For the analysis of ADMA, SDMA, L-NMMA, arginine, homoarginine, citrulline, ornithine, 200 Âµl of serum were precipitated with methanol, and subsequently derivatized with a butanol solution containing 5% acetyl chloride. Butyl derivatives were separated using a C18 reverse phase column with a 5 min run time. Detection of analytes was achieved by utilising the specific fragmentation patterns identified through tandem mass spectrometry., Results: The method was linear for ADMA, SDMA, L-NMMA, ornithine, arginine, homoarginine and citrulline in the ranges of 0.023-6.0, 0.021-5.5, 0.019-5.0, 0.015-250, 0.015-250, 0.019-5 and 0.015-250 µM, respectively. The inter-assay CV% values for all analytes was less than 9.8%., Conclusions: Data obtained from method validation studies shows that the developed method is highly sensitive, precise and accurate. Short analysis time, cost-effectiveness, and multiplexed analysis of these metabolites, with the same pretreatment steps, are the main advantages of the method., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 THE AUTHORS. Publishing services by ELSEVIER B.V. on behalf of MSACL.)

Background: Acne vulgaris is a common skin disease in adolescents known to be associated with oxidative stress. However, the number of studies in which oxidative stress and antioxidants are evaluated together is limited., Aims: In this study, we aimed to investigate L-arginine/nitric oxide (NO) pathway metabolites, ischemia-modified albumin (IMA), and vitamin A and E levels in patients with acne and its association with disease severity., Patients/methods: Ninety patients with acne and 30 healthy adults were included in the study. The serum levels ofL-arjinin, L-arginine metabolites, IMA, and vitamins A and E measured in the patient and control groups., Results: Asymmetric dimethylarginine (ADMA), LN G -monomethyl-L-arginine (L-NMMA) and symmetric dimethylarginine (SDMA), and IMA levels were significantly higher in the patients with acne than in the control group (P ˂ .05). The L-arginine/ADMA ratio and citrulline and vitamin A levels were significantly lower in patients with acne than those of the controls (P ˂ .05). ADMA and IMA plasma levels were increased in parallel with the disease severity (P ˂ .05). L-arginine/ADMA ratio, L-arginine, citrulline, and vitamin A plasma levels decreased as the disease became severe (P ˂ .05). Although L-arginine and vitamin E levels were lower in the patient group compared to the control group, the difference was not statistically significant (P ˃ .05)., Conclusion: These results suggest that IMA and L-arginine-NO pathway associated with ischemia and oxidative stress may play an important role in the pathogenesis and progression of acne vulgaris., (© 2020 Wiley Periodicals, LLC.)

Objective: Clozapine is one of the most effective drugs for resistant schizophrenia, but its severe metabolic and hematological side effects limit the use of clozapine. It has been reported that clozapine blood concentrations should be maintained between 350-600 ng/mL. Our aim was to develop a determination method for clozapine and its main metabolites norclozapine and clozapine-N-oxide, to perform validation studies and to investigate the change of various biochemical parameters in patients using clozapine., Methods: A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for clozapine measurement. Thus, blood samples were collected from 38 patients with schizophrenia and 32 healthy volunteers. Biochemical and hematological parameters were measured by Beckman-Coulter AU 5800 (Beckman Coulter, Brea, USA) and Beckman Coulter LH 780 analyzer (Beckman Coulter, Miami, FL, USA), respectively. Hormone levels were analyzed using Cobas 6000 analyzer (Roche Diagnostics, Germany)., Results: The LCMS/MS method was linear between 1.22-2500 ng/mL (r 2  = 0.9971) for clozapine. The retention times of clozapine, norclozapine and clozapine-N-oxide were 0.92, 0.89 and 0.95, respectively. Blood glucose (GLU) (p = 0.025), low density lipoprotein (LDL-cholesterol) (p = 0.015), triglyseride (TG) (p = 0.042) and total cholesterol (TC) (p = 0.024) levels were higher; hemoglobin (HGB) (0.015), mean corpuscular hemoglobin (MCH) (0.036), red blood cell count (RBC) (0.020), neutrophil (NEU) (0.034), and platelet (PLT) (P = 0.005) levels were lower in the clozapine group., Conclusions: This LC-MS/MS method was rapid, simple, cost-effective and suitable for the routine clozapine monitoring. Furthermore, norclozapine and clozapine-N-oxide were also determined. Monitoring of metabolic and hematological parameters with clozapine levels is very important. However, the limitations of the study were that the method was not validated for norclozapine and clozapine-N-oxide, so the validation parameters were not evaluated for these two metabolites., Competing Interests: Declaration of Competing Interest None., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Asymmetric dimethylarginine, symmetric dimethylarginine, and L-monomethylarginine are originated from the subsequent proteolytic catalysis of methylated arginine residues on different proteins and inhibit the endogenous nitric oxide generation. The changes in total methylarginine load (Asymmetric dimethylarginine plus symmetric dimethylarginine plus L-monomethylarginine) may contribute to hypertension. The aim of this study was to determine serum methylarginine concentrations in patients with masked hypertension and determine the association between these biomarkers and blood pressure measurements. Control group, masked hypertension and hypertension groups consisted of 40 subjects (11 males, 28 females, mean age 48.6 ± 13.1), 28 subjects (14 males, 14 females, mean age 50.9 ± 11.0) and 36 subjects (15 males, 21 females, mean age 54.4 ± 12.3 years), respectively ( P = 0.149). Serum total methylarginine load was significantly higher in hypertension group (0.63 ± 0.23) compared to masked hypertension (0.49 ± 0.16) and control groups (0.38 ± 0.13) ( P = 0.008 and P < 0.001). While there was no statistically significant difference between healthy control groups [0.147 (0.03-0.29)] and masked hypertension patients [0.144 (0.05-0.42)] for serum symmetric dimethylarginine levels ( P = 0.970), it was markedly elevated in hypertension group [0.25 (0.07-0.54)] compared to masked hypertension group [0.14 (0.05-0.42)] ( P = 0.001). Serum total methylarginine load was positively correlated with night-time SBP (r = 0.214, P = 0.029). Serum methylarginine levels might be a useful marker for determining the courses of clinical hypertension.

Background: Depression is common in chronic kidney disease (CKD) patients and associated with significant increase in morbidity and mortality. In recent years, a relationship between vitamin D deficiency and depression has been shown. The aim of this study is to investigate the relationship between 25-hydroxy (OH) vitamin D and depression in hemodialysis patients., Methods: A total of 140 patients were included in the study. Hamilton depression scale (HAM-D) was completed by all patients. 25(OH) vitamin D levels were compared between patients with and without depressive symptoms., Results: Patients who had depressive symptoms had significantly lower 25(OH) vitamin D levels (13.70 [24.3-8.25] vs. 18.20 [29.2-11.7] ng/mL, p = 0.016). HAM-D score showed significant association with gender (p = 0.011) and 25(OH) vitamin D level (p = 0.011). Univariate logistic regression analysis showed that males had lower risk of depression by a ratio of 61.1% (OR 0.389, p = 0.012) and vitamin D-deficient patients had 2.88 times greater risk of depression compared to non-deficient patients (OR 2.885, p = 0.013). Multivariate logistic regression analysis showed that males had 53.7% less risk of depression (OR 0.463, p = 0.046) and vitamin D-deficient patients had 2.39 times greater risk of depression (OR 2.397, p = 0.047). When evaluated by gender, univariate logistic regression analysis showed that 25(OH) vitamin D and other variables were not associated with depression in females (p > 0.05), while only vitamin D level had a significant effect on depression in males (OR 8.207, p = 0.008)., Conclusions: We found a significant association between vitamin D level and depressive symptoms in hemodialysis patients. When analyzed according to gender, this association was found to stand independent of other variables only in males.

Introduction: Imatinib has favorable pharmacokinetic properties, but primary and secondary resistance mechanisms may cause a decrease in clinical response over time. There is a positive correlation between serum imatinib concentrations and treatment response. Our aim was to develop a method for the measurement of imatinib and its' active metabolite N-desmethyl imatinib., Methods: Serum imatinib and N-desmethyl imatinib levels were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and validation studies were carried out according to CLSI (The Clinical & Laboratory Standards Institute) protocols. Serum samples were collected from 40 patients with chronic myeloid leukemia (CML) and analyzed with LC-MS/MS and ultra high-performance liquid chromatography (UHPLC) methods., Results: The linearity range and correlation coefficient were 12.2-12,500 ng/mL and 0.9987 for LC-MS/MS method, respectively. Limit of quantitation was determined as 24.4 ng/mL. The retention times of imatinib and N-desmethyl imatinib were 1.66 and 1.60 min, respectively. There was no statistically significant difference between the results of both methods., Discussion: This LC-MS/MS method is cost-effective and has adavantages such as using low serum volumes, requiring simple pretreatment steps (only protein precipitation) and reduced turnaround times for analysis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Nitric oxide synthase (NOS) is present in the brain and cerebral arteries and it enables the synthesis of nitric oxide (NO), which plays a critical role in brain perfusion. Asymmetrical dimethylarginine (ADMA) is an endogenous NOS inhibitor., Objectives: The aim of this study was to evaluate serum ADMA levels, which are an indicator of endothelial dysfunction of the renal functions in patients with acute ischemic stroke, and to determine whether there is a possible correlation between ADMA and NO levels and the l-arginine-to-ADMA ratio., Material and Methods: Fifty-two patients (22 male and 30 female; mean age: 75.2 ±10.1 years) with a diagnosis of acute ischemic stroke in the first 24 h post-stroke and 48 healthy individuals (controls; 13 male and 35 female; mean age: 60.1 ±7.92 years) were included in this study. The risk factors recorded and evaluated were age and gender of the patients, serum lipid levels, serum ADMA levels, nitrate-to-nitrite ratios, l-arginine, l-arginine-to-ADMA ratios, sedimentation rate, C-reactive protein (CRP), urea and creatinine levels, and glomerular filtration ratio (eGFR)., Results: The mean serum ADMA level was 0.48 ±0.23 μM for the patients and 0.36 ±0.18 μM for the controls. The mean NO level was 2.78 ±0.59 μM for the patient group and 4.49 ±2.84 μM for the controls. The ADMA levels for the patient group were significantly higher than for the control group (p = 0.011); the NO levels for the patients were significantly lower than for the controls (p < 0.001). The logistic regression method demonstrated that ADMA and NO levels may be independent risk factors for the patient group, and the receiver operating characteristic (ROC) curve analysis showed that both of these variables were discriminative risk factors., Conclusions: An increased serum level of the NOS inhibitor ADMA was found to be a possible independent risk factor for ischemic stroke.

A decrease in bone mass is observed in hemophilic patients. The aim of this study was to evaluate bone mineral density (BMD), parathyroid hormone (PTH), 25-hydroxy vitamin D (vitamin D), and a bone formation and resorption marker, procollagen type I N-terminal propeptide (PINP) and urinary N-terminal telopeptide (uNTX) respectively, in hemophilic patients and healthy controls. Laboratory parameters related to the pathogenesis of bone loss such as neutrophil-lymphocyte ratio (NLR) and platelet-lymphocyte ratio (PLR) were also evaluated. Thirty-five men over 18 years of age, with severe hemophilia (A and B) and receiving secondary prophylaxis, were included in the study. The same number of age-, sex-, and ethnicity-matched healthy controls were evaluated. Anthropometric, biochemical, and hormonal parameters were determined in both groups. No significant difference in anthropometric parameters was found between the two groups. The BMD was low in 34% of hemophilic patients. Vitamin D, calcium, and free testosterone levels were significantly lower (p < 0.001, p = 0.011, p < 0.001, respectively), while PTH, PINP, and activated partial thromboplastin time (aPTT) levels were significantly higher (p < 0.014, p = 0.043, p < 0.001, respectively), in hemophilic patients compared to controls. There was no significant difference between the two groups in NLR, PLR, phosphorus, thyroid-stimulating hormone, and uNTX level. The reduction of bone mass in hemophilic patients may be evaluated using the markers of bone formation and resorption, enabling early detection and timely treatment.

Objective: The objective of this study is to compare serum levels of FKN and SFRP-4 in patients with normal glucose tolerance (NGT), impaired glucose tolerance (IGT), and type 2 diabetes mellitus (T2DM)., Methods: A total of 152 patients presented to the endocrinology outpatient clinic of our hospital were included in the study. Eighty-two patients with a history of T2DM were assigned to the T2DM group. IGT (n = 34) and NGT (n = 36) groups included the patients who received oral glucose tolerance test outcomes., Results: Serum FKN levels were significantly higher in the IGT and T2DM groups compared to the NGT group (p < 0.001 and p < 0.001, respectively). Serum SFRP-4 levels were significantly higher in the T2DM group compared to the IGT and NGT groups (p = 0.001 and p = 0.004, respectively). A significant correlation was observed between FKN and fasting glucose levels. SFRP-4 was significantly correlated with fasting glucose, HbA1c, and triglyceride levels., Conclusion: To our knowledge, increased FKN levels in patients with IGT were demonstrated for the first time in this study. The results of our study support the opinion that FKN and SFRP-4 may contribute to the pathogenesis of T2DM (Tab. 1, Fig. 3, Ref. 23).

Background: Recent reports have described inherent problems with androgen immunoassays compared with mass spectrometry analyses. In this study, a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed according to CLSI rules. The developed method was compared with two immunoassay methods, the enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA)., Methods: After liquid-liquid extraction, a Shimadzu Prominence LC unit coupled to an ABSCIEX API 3200 mass spectrometer with atmospheric pressure chemical ionization was used to quantify serum androstenedione levels. Serum androstenedione results taken from tandem mass spectrometry were compared with the immunoassays., Results: The androstenedione assay was linear up to 50 ng/mL. Lower limit of quantitation and lower limit of detection were 0.195 ng/mL and 0.097 ng/mL, respectively. This method was not affected by matrix effect and other steroid hormone interferences. In this study, the obtained recovery was 69 - 99%, carryover value was determined as 0.026 ng/mL. According to the results of an interference study, androstenedione bias % did not exceed the limit of allowable bias % and 88.7% recovery was acquired for androstenedione. In the LC-MS/MS and ELISA comparison study, the slope value was found as 18.412, intercept -22.87, and r2 value as 0.1033. In the LC-MS/MS and RIA comparison study, slope value was found as 1.085, intercept 0.4541, and r2 value as 0.3712. In the RIA and ELISA comparison study, slope value was found as 9.57, intercept -15.5, and r2 value as 0.19., Conclusions: The LC-MS/MS provides agreement with the results of radioimmunoassay but not with ELISA. This method offers better selectivity compared to immunoassay systems.

The role of vitamin D in calcium absorption and bone health is known. The studies revealed that vitamin D modulates breast cancer cell growth and it is also associated with a reduced breast cancer risk. The primary objective of this study was to highlight the metabolic effect of Vitamin D on MCF-7 breast cancer cell line. For that purpose, we checked the apoptosis, energy, amino-acid and acylcarnitine levels in cancer cells, that the study propose, that 1α, 25(OH)2D3 could inhibit cell growth in a dose and time dependent manner. IC50 dose was calculated as 145 nM for vitamin D. We observed the apoptosis level in vitamin D groups, which were 18, 28 and 38.5 % at 24, 48 and 72 hours, respectively. During metabolic screening analysis, it was observed that glutamine, methionine and glutamic acid levels were treated more by Vitamin D groups in cell line and also, that acylcarnitine level was increased in 24 and 48 hour groups when compared to the control, but decreased in 72 hours. Further studies are needed to analyze the role of amino acids and acylcarnitines for early apoptosis and cancer metabolism (Tab. 2, Fig. 4, Ref. 24).