End-user verification results of two serum separator tubes for clinical chemistry analytes according to CLSI GP34-A and CLSI GP41-A6.

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.

Comparison of Labsan Tricell-1000 and Dirui FUS-2000 automated urine analyzers with manual microscopy.

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 (Ï° = 0.80), and WBC (Ï° = 0.83); good for CaOx (Ï° = 0.69), SEC (Ï° = 0.80), YLC (Ï° = 0.72), HC (0.69) and LC (Ï° = 0.64); moderate for BAC (Ï° = 0.51), APC (Ï° = 0.43) and MT (Ï° = 0.55); fair for GC (Ï° = 0.39) and RTEC (Ï° = 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.

Comparison of scoring systems for predicting short- and long-term type 2 diabetes remission after bariatric surgery.

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.

Evaluation of Bone Turnover Markers Such as Osteoprotegerin, Sclerostin and Dickkopf-1 in Subclinical Hyperthyroidism.

In this study, it was aimed to assess effects of subclinical hyperthyroidism (SH) on bone metabolism using osteoprotegerin (OPG), sclerostin, Dickkopf-1 (DKK1) and biochemical parameters. This cross-sectional prospective study included 40 patients with SH and 40 euthyroid controls. Serum OPG, sclerostin, DKK-1, type-1 procollagen, C-terminal polypeptide (CTx) and 24-hours urine N-terminal telopeptide (NTx) were measures using ELISA kit. Bone mineral density measurements were performed using dual energy X-ray absorptiometry (DEXA). Risk for 10-years hip and major fracture was estimated by Turkish version of FRAX. No significant difference was detected in age, gender, body mass index, smoking and menopause rates between SH and control groups. The risk for 10-years hip fracture and major osteoporotic fracture were estimated as 4.45% and 0.55% in SH group, respectively. The OPG levels were significantly lower in patients with SH than controls (P = 0.017). No significant difference was detected in other bone formation and degradation parameters. No significant correlation was detected between OPG level and risk for major osteoporotic fracture (P > 0.05); however, a negative correlation was detected between OPG level and risk for hip fracture (rho = 0.233; P = 0.038). Serum OPG is markedly affected in patients with SH. In addition, OPG seemed to be associated with osteoporotic fracture risk. Available data show that SH is significantly associated with risk for fracture; thus, it is important to assess risk for fracture in patients with SH.

Comparison of three chatbots as an assistant for problem-solving in clinical laboratory.

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.

Relationship of tryptophan metabolites with the type and severity of multiple sclerosis.

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.

Measurement of kynurenine pathway metabolites by tandem mass spectrometry.

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.

Impaired kynurenine metabolism in patients with primary Sjögren's syndrome.

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.

The Role of Stem Cells Derived From the Mesenchyme of the Umbilical Cord in Reducing Immunosuppressive Drug Doses Used in Allogenic Transplantations.

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.

Assessment of the diagnostic performance of the 1 mg dexamethasone suppression test in class 3 obese patients.

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.

Comparison of scoring systems for predicting remission of type 2 diabetes in sleeve gastrectomy patients.

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.

Raised total methylated arginine load in patients with gout.

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.

Quantification of serum homoarginine, methylated arginine and inhibin-A levels in a high-risk pregnancy.

The plasma levels of homoarginine (h-Arg) and methylated arginine have proven to be an independent cardiovascular risk factor. We aimed to determine the h-Arg and methyl arginine levels in serums of high-risk pregnancy causing potential complications. These participants were divided into four groups as the control group with quadruple test, the high-risk group quadruple test the control group with binary test, the high-risk group with quadruple test that have a positive result from second-trimester screening with a cut-off value of 1 in 300. The serum methyl arginine and homoarginine levels were analysed with liquid chromatography-tandem mass spectrometry. Serum h-Arg levels were found to be higher in high-risk groups compared to control groups and it was also detected higher in the groups with quadruple test than the groups with binary test (p < .05). H-Arg levels in the groups showed strong negative correlation with age and serum inhibin-A levels (r = -0.288, p < .001). Also, there was a strong negative correlation between serum asymmetric dimethylarginine (ADMA) and serum inhibin-A levels (r = -0.352, p < .001). H-Arg may be a new risk marker to detect high-risk pregnancies in early pregnancy. In addition to, methylated arginine such as ADMA has a key regulator in a physiological concentration of h-Arg.IMPACT STATEMENTWhat is already known on this subject? H-Arg levels decrease may be associated with preeclampsia, GDM, macrosomia, low birth weight, and preterm delivery in pregnancy.What do the results of this study add? Serum h-Arg levels were found to be higher in high-risk groups. Additionally, h-Arg levels and ADAM, one of the methylated arginines in the groups showed a strong negative correlation with serum inhibin-A levelsWhat are the implications of these findings for clinical practice and/or further research? H-Arg may be a new risk marker to detect high-risk pregnancies.

Elevated serum levels of kynurenine pathway metabolites in patients with Behçet disease.

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.

New steps in acute coronary syndrome and antihyperlipidemic treatment: Determination of statins and metabolites by liquid chromatography tandem mass spectrometry.

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.

Effect of calcium and vitamin D supplementation on the clinical, hormonal, and metabolic profile in non-obese women with polycystic ovary syndrome.

BACKGROUND: In this study, we investigated the effect of calcium and vitamin D (Ca/Vit D) supplementation on the clinical, hormonal, and metabolic profile of patients with low vitamin D levels. In addition, we investigated the effect of Ca/Vit D supplementation on asymmetric dimethylarginine (ADMA) level in patients with polycystic ovary syndrome (PCOS). METHODS: In total, 75 patients aged 19-35 years, with a normal body mass index and a diagnosis of PCOS and Vit D deficiency/insufficiency, were included in the study. Patients received 50,000 IU of vitamin D3 once a week for 8 weeks. Afterward, 2500 mg calcium carbonate equivalent to 1000 mg calcium ion and 9.68 mg cholecalciferol equivalent to 880 IU vitamin D3 were administered orally as a maintenance treatment once a day. RESULTS: The mean age of the patients was 21.7 ± 3.5. After Ca/Vit D supplementation, Vit D levels significantly increased compared to baseline (8.6 ng/ml) levels. An increase in SHBG levels (p < 0.001), a decrease in total testosterone, FAI (p = 0.042), and ADMA levels (p < 0.001) were observed in the first and third months compared to the onset. Significant improvement compared to baseline was observed in menstrual irregularity and median mFG score. CONCLUSION: Ca/Vit D supplementation can improve PCOS symptoms such as menstrual dysfunction, hirsutism, and hyperandrogenism. It may be effective in reducing the risk of cardiovascular disease in patients with PCOS later in life by decreasing ADMA levels, which is an indicator of endothelial dysfunction.

Altered kynurenine pathway metabolite levels in toddlers and preschool children with autism spectrum disorder.

AIM: There are increasing reports about the potential role of kynurenine pathway metabolites in autism spectrum disorder (ASD). Early childhood is a very crucial period of time for the etiopathogenesis of ASD and previous studies reported an age-dependent alteration in kynurenine metabolism. However, no study specifically examined kynurenine metabolites in very young children with ASD. This study aimed to investigate kynurenine pathway metabolite levels, kynurenine pathway enzyme activities and neuroprotective index (kynurenic acid/3-hydroxykynurenine ratio) in toddlers and preschool children with ASD. MATERIALS AND METHODS: A total of 68 children with ASD and 44 healthy controls aged between 18 and 60 months were included in this study. Serum levels of kynurenine pathway metabolites were determined by liquid chromatography-mass spectrometry/mass spectrometry system. RESULTS: Serum 3-hydroxykynurenine and kynurenic acid concentrations were significantly higher in the ASD group than in the control group, whereas serum 3-hydroxyanthranilic acid concentrations were significantly lower. CONCLUSIONS: These findings showed that the kynurenine pathway may play a role in the etiopathogenesis of ASD in early childhood.

A study on relationship between androgenetic alopecia and cardiovascular risk using high sensitivity C-reactive protein and galectin-3 in men with early onset baldness.

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.

Developing a robust, fast and reliable measurement method for the analysis of methylarginine derivatives and related metabolites.

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.

Effects of hydroxychloroquine and its metabolites in patients with connective tissue diseases.

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.