Tissue and plasma free amino acid detection by LC-MS/MS method in high grade glioma patients.

PURPOSE: The changes in serum amino acid profiles are evaluated in different types of cancers and screening tests were developed for estimating the risk of cancer by rapid analysis of plasma free amino acid (PFAA) levels. There is scarce evidence about the metabolomics analysis of PFAA in malignant gliomas. The aim of the present study was to identify the most promising diagnostic amino acid biomarkers that could be objectively measured for high-grade glioma and to compare their level with the tissue counterpart. METHODS: In this prospective study, we collected serum samples from 22 patients with the pathological diagnosis of high-grade diffuse glioma according to WHO 2016 classification and 22 healthy subjects, and brain tissue from 22 controls. Plasma and tissue amino acid concentrations were analyzed applying liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. RESULTS: Serum alanine, alpha-aminobutyric acid (AABA), lysine (Lys) and cysteine concentrations were significantly higher in high-grade glioma patients despite low levels of alanine and Lys in the tumor tissue. Aspartic acid, histidine and taurine were significantly decreased in both serum and tumors of glioma patients. A positive correlation was detected between tumor volumes and serum levels of latter three amino acids. CONCLUSION: This study demonstrated potential amino acids which may have diagnostic value for high-grade glioma patients by utilizing LC-MS/MS method. Our results are preliminary to compare serum and tissue levels of amino acids in patients with malignant gliomas. The data presented here may provide feature ideas about the metabolic pathways in the pathogenesis of gliomas.

Within- and between-subject biological variation data for serum zinc, copper and selenium obtained from 68 apparently healthy Turkish subjects.

OBJECTIVES: Trace elements (TrEL) are nutritionally essential components in maintaining health and preventing diseases. There is a lack of reliable biological variation (BV) data for TrELs, required for the diagnosis and monitoring of TrEL disturbances. In this study, we aimed to provide updated within- and between-subject BV estimates for zinc (Zn), copper (Cu) and selenium (Se). METHODS: Weekly serum samples were drawn from 68 healthy subjects (36 females and 32 males) for 10 weeks and stored at -80 °C prior to analysis. Serum Zn, Cu and Se levels were measured using inductively-coupled plasma mass spectrometry (ICP-MS). Outlier and variance homogeneity analyses were performed followed by CV-ANOVA (Røraas method) to determine BV and analytical variation estimates with 95% CI and the associated reference change values (RCV) for all subjects, males and females. RESULTS: Significant differences in mean concentrations between males and females were observed, with absolute and relative (%) differences for Zn at 0.5 µmol/L (3.5%), Cu 2.0 µmol/L (14.1%) and Se 0.06 µmol/L (6.0%). The within-subject BV (CVI [95% CI]) estimates were 8.8% (8.2-9.3), 7.8% (7.3-8.3) and 7.7% (7.2-8.2) for Zn, Cu and Se, respectively. Within-subject biological variation (CVI) estimates derived for male and female subgroups were similar for all three TrELs. Marked individuality was observed for Cu and Se. CONCLUSIONS: The data of this study provides updated BV estimates for serum Zn, Cu and Se derived from a stringent protocol and state of the art methodologies. Furthermore, Cu and Se display marked individuality, highlighting that population based reference limits should not be used in the monitoring of patients.

Rapid detection of serum free light chains by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Introduction: Serum free light chain (FLC) measurements are increasingly prominent for patients with plasma cell disorders (PCDs) in screening, prognostic stratification, and monitoring therapy responses. Objectives: We aimed to develop a sensitive, reliable, and accurate method for diagnosing PCDs that can notably decrease the time and cost of current methods. Methods: Here, we present a novel approach for FLC measurement using immunoenrichment on micro-affinity chromatography in combination with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) detection. In this study, serum free kappa (κ) and free lambda (λ) light chain (LC) levels in the serum of 105 patients were compared between the nephelometric serum FLC quantification and MALDI-TOF MS detection. Results: Cohen's kappa coefficient between the MALDI-TOF MS-based method and the FLC assay revealed an almost perfect agreement in the case of normal (negative) results (κ = 0.92; 95% confidence interval (CI): 0.837 to 0.968) and a good agreement in the case of increased (positive) results (κ = 0.76; 95% CI: 0.608 to 0.870). In Spearman's correlation analysis, the best correlation was found between serum free κ/λ ratios (r = 0.628, 0.496 to 0.732; p <0.0001). Our method showed sensitivity (92.5%) and specificity (76.3%) for discrimination between the κ/λ FLC ratio compared to the serum FLC assay. Conclusion: The proposed method can significantly contribute to diagnosing and monitoring PCDs as it can significantly be time-saving, cost-effective in FLC measurement.

Melatonin with 1,25-dihydroxyvitamin D3 protects against apoptotic ischemia-reperfusion injury in the rat kidney.

This study was designed to evaluate the preventive role of melatonin (Mel) and 1,25-dihydroxyvitamin D3 (VD3) in biochemical and apoptotic events leading to tissue injury and renal dysfunction after ischemia-reperfusion (I/R). Thirty male Wistar rats were divided into five groups: sham-operated, I/R, Mel + I/R, VD3 + I/R, and Mel + VD3 + I/R. The rats were intraperitoneally administered with Mel (10 mg/kg), VD3 (0.5 µg/kg), or Mel (10 mg/kg) plus VD3 (0.5 µg/kg) each day at 1 week prior to ischemia. Right nephrectomy was initially performed and left renal I/R injury was induced by 45 min of bilateral renal ischemia followed by 45 min of reperfusion. After reperfusion, kidneys and blood were obtained for histopathologic and biochemical evaluation. Mel and VD3 had an ameliorative effect on biochemical parameters such as serum creatinine, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, and apoptosis (caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining) in the kidneys against renal I/R injury in rats. Additionally, VD3 combined with Mel significantly reduced apoptotic and histological alterations when compared with Mel or VD3 alone. This preventive effect on renal tubular apoptosis was remarkable when Mel was combined with VD3.

Leukocyte telomere length as a compensatory mechanism in vitamin D metabolism.

Vitamin D deficiency is common among postmenopausal women. Telomere length can be a potential protective mechanism for age-related diseases. The objective of our study is to examine the association of vitamin D supplementation on leukocyte telomere length (LTL) in healthy postmenopausal women with vitamin D deficiency. The study was designed as a placebo-controlled study to investigate the short-term effects of vitamin D supplementation and seasonal changes on vitamin D related parameters, including 25(OH)D, 1,25(OH)2D parathormone (PTH), Vitamin D binding protein (VDBP), vitamin D receptor (VDR), and telomere length in a cohort of postmenopausal women (n = 102). The group was divided as supplementation (n = 52) and placebo groups (n = 50). All parameters were measured before and after treatment. Serum VDBP levels were measured by ELISA method and VDR, GC (VDBP) gene expressions and relative telomere lengths were measured in peripheral blood mononuclear cells (PBMC) using a quantitative real-time PCR method. The results demonstrate that baseline levels were similar between the groups. After vitamin D supplementation 25(OH)D, 1,25(OH)2D, PTH and VDBP levels were changed significantly compared to the placebo group. At the end of the study period, LTL levels were significantly increased in both groups and this change was more prominent in placebo group. The change in GC expression was significant between treatment and placebo groups but VDR expression remained unchanged. Even though the study was designed to solely assess the effects of vitamin D supplementation, LTL was significantly increased in the whole study group in summer months suggesting that LTL levels are affected by sun exposure and seasonal changes rather than supplementation. The study displayed the short-term effect of Vitamin D supplementation on vitamin D, PTH levels, LTL and vitamin D associated gene expressions. The relation between Vitamin D and LTL is not linear and could be confounded by several factors such as the population differences, regional and seasonal changes in sun exposure.

Effects of Prolonged Intermittent Fasting Model on Energy Metabolism and Mitochondrial Functions in Neurons.

Background: Calorie restriction (CR) during daily nutrition has been shown to affect the prognosis of many chronic diseases such as metabolic syndrome, diabetes, and aging. As an alternative nutrition model, prolonged intermittent fasting (PF) in humans is defined by the absence of food for more than 12 h. In our previous human studies, CR and PF models were compared and it was concluded that the two models might have differences in signal transduction mechanisms. We have investigated the effects of these models on neurons at the molecular level in this study. Methods: Neurons (SH-SY5Y) were incubated with normal medium (N), calorie-restricted medium (CR), fasting medium (PF), and glucose-free medium (G0) for 16 h. Simultaneously, ketone (beta-hydroxybutyrate; bOHB) was added to other experiment flasks containing the same media. Concentrations of lactate, lactate dehydrogenase (LDH), bOHB, and glucose were measured to demonstrate the changes in the energy metabolism together with the mitochondrial functions of cells. Citrate synthase activity and flow cytometric mitochondrial functions were investigated. Results: At the end of incubations, lactate and LDH levels were decreased and mitochondrial activity was increased in all ketone-added groups (P < .01) regardless of the glucose concentration in the environment. In the fasting model, these differences were more prominent. Conclusion: Our results demonstrated that neurons use ketones regardless of the amount of glucose, and bOHB-treated cells had positive changes in mitochondrial function. We conclude that the presence of bOHB might reverse neuron damage and that exogenous ketone treatment may be beneficial in the treatment of neurological diseases in the future.

Kappa/Lambda light-chain typing in Alzheimer's Disease.

BACKGROUND: Alzheimer's disease is a progressive neurodegenerative disorder characterized by memory loss and cognitive impairment. The diagnosis of Alzheimer's disease according to symptomatic events is still a puzzling task. Developing a biomarker-based, low-cost, and high-throughput test, readily applicable in clinical laboratories, dramatically impacts the rapid and reliable detection of the disease. OBJECTIVE: This study aimed to develop an accurate, sensitive, and reliable screening tool for diagnosing Alzheimer's disease, which can significantly reduce the cost and time of existing methods. METHODS: We have employed a MALDI-TOF-MS-based methodology combined with a microaffinity chromatography enrichment approach using affinity capture resins to determine serum kappa (κ) and lambda (λ) light chain levels in control and patients with AD. RESULTS: We observed a statistically significant difference in the kappa light chain over lambda light chain (κLC/λLC) ratios between patients with AD and controls (mean difference -0,409; % 95 CI:- 0.547 to -0.269; p<0.001). Our method demonstrated higher sensitivity (100.00%) and specificity (71.43%) for discrimination between AD and controls. CONCLUSION: We have developed a high-throughput screening test with a novel sample enrichment method for determining κLC/λLC ratios associated with AD diagnosis. Following further validation, we believe our test has the potential for clinical laboratories.

Bronchial hyperreactivity and airway wall thickening in obstructive sleep apnea patients.

PURPOSE: Hypoxia/reoxygenation episodes in obstructive sleep apnea (OSA) results in the alteration of the oxidative balance, leading to the development of inflammation. Airway wall thickening and inflammatory changes are suggested as a primary cause of the airway hyperresponsiveness in asthmatics. Bronchial hyperreactivity (BH) may also occur in patients with OSA. We investigated the presence of BH and airway wall thickness in OSA and correlations with inflammatory markers. MATERIALS AND METHODS: Sixteen OSA patients and ten controls without allergic diseases were prospectively studied. Plasma pro-B-type natriuretic peptide (pro-BNP), fibrinogen, D-dimer, α1-antitrypsin, and high-sensitive C-reactive protein levels were measured. Airway wall thickness was evaluated with high-resolution CT, and BH was assessed by giving each subject a methacholine challenge test. RESULTS: In OSA patients, bronchial wall thickness, fibrinogen, D-dimer, α1-antitrypsin, high sensitive C-reactive protein, and pro-BNP levels were significantly greater than those in control subjects. Among the 16 patients, three had BH on methacholine challenge. Bronchial wall thickness(mm) was positively correlated with apnea-hypopnea index (AHI: number of apneas + hypopneas/hour of sleep), BMI, respiratory arousal index, nocturnal oxygen desaturation (NOD) duration (time in minutes with a nocturnal arterial oxygen saturation of <90% during sleep), and α1-antitrypsin levels. NOD duration also correlated with pro-BNP and fibrinogen levels. CONCLUSIONS: In OSA patients, walls of central airways were thicker than normal subjects. BH may have occurred in OSA patients. NOD duration correlated with inflammatory parameters and oxygen desaturation index 3% had an effect on the thickness of bronchial walls. But overall, AHI was found to be the only independent predictor of bronchial wall thickness.

Early Postnatal Metabolic Profile in Neonates With Different Birth Weight Status: A Pilot Study.

Introduction: Restricted or enhanced intrauterine growth is associated with elevated risks of early and late metabolic problems in humans. Metabolomics based on amino acid and carnitine/acylcarnitine profile may have a role in fetal and early postnatal energy metabolism. In this study, the relationship between intrauterine growth status and early metabolomics profile was evaluated. Materials and Methods: A single-center retrospective cohort study was conducted. Three hundred and sixty-one newborn infants were enrolled into the study, and they were grouped according to their birth weight percentile as small for gestational age (SGA, n = 69), appropriate for gestational age (AGA, n = 168), and large for gestational age (LGA, n = 124) infants. In all infants, amino acid and carnitine/acylcarnitine profiles with liquid chromatography-tandem mass spectrometry (LC-MS/MS) were recorded and compared between groups. Results: LGA infants had higher levels of glutamic acid and lower levels of ornithine, alanine, and glycine (p < 0.05) when compared with AGA infants. SGA infants had higher levels of alanine and glycine levels when compared with AGA and LGA infants. Total carnitine, C0, C2, C4, C5, C10:1, C18:1, C18:2, C14-OH, and C18:2-OH levels were significantly higher and C3 and C6-DC levels were lower in SGA infants (p < 0.05). LGA infants had higher C3 and C5:1 levels and lower C18:2 and C16:1-OH levels (p < 0.05). There were positive correlations between free carnitine and phenylalanine, arginine, methionine, alanine, and glycine levels (p < 0.05). Also, a positive correlation between ponderal index and C3, C5-DC, C14, and C14:1 and a negative correlation between ponderal index and ornithine, alanine, glycine, C16:1-OH, and C18:2 were shown. Conclusion: We demonstrated differences in metabolomics possibly reflecting the energy metabolism in newborn infants with intrauterine growth problems in the early postnatal period. These differences might be the footprints of metabolic disturbances in future adulthood.

Attenuated lactate response to ischemic exercise in migraine.

BACKGROUND: Although the pathogenesis of migraine still remains unclear, certain metabolic studies done on patients with migraine indicate possible deficits in mitochondrial activity. Previously, the forearm ischemic exercise test (FIT) has been used as a screening tool to evaluate mitochondrial dysfunction in metabolic myopathies. MATERIAL/METHODS: We studied the response of migraine patients to exercise using the modified FIT and compared this to the responses in a healthy group. After baseline venous blood samples were drawn, a sphygmomanometer cuff placed around the upper arm was inflated to 30 mmHg above systolic blood pressure and the subject performed a maximal isometric contraction of the forearm flexors using a hand-grip dynamometer with a 9-contraction: 1-relaxation duty cycle for a total time of 60 seconds. RESULTS: In migraine patients, attenuated lactate response after the FIT in the 1st, 3rd and 5th minutes were significantly different when compared to the control group. CONCLUSIONS: Our results show that there may be defects in the anaerobic or proximal glycolytic pathways in migraine patients, evident in stressful situations. We also conclude that the FIT may be useful for research on migraine pathophysiology.

Homocysteine levels and echocardiographic findings in obstructive sleep apnea syndrome.

BACKGROUND: The obstructive sleep apnea syndrome (OSAS) is characterized by repeated upper airway obstruction during sleep together with decreases in oxygen saturation leading to a series of pathological events, primarily in the cardiovascular system. Elevated plasma homocysteine levels have recently been considered as an independent risk factor for vascular disease, and increased levels are attributed to cardiovascular diseases. OBJECTIVES: We aimed to investigate the possible relationship between homocysteine levels and echocardiographic findings in OSAS patients at different stages of disease. METHODS: Thirty-eight patients (23 males and 15 females) with polysomnographically verified OSAS (mean age, 49 +/- 12 years, range 27-74) and a mean body mass index of 31.27 +/- 5.24 kg/m(2) (range 22.60-47.90) were prospectively studied. Plasma levels of homocysteine, cholesterols, triglycerides, vitamin B(12) and high-sensitive C-reactive protein (hsCRP), as well as echocardiographic and lung function parameters were assessed. RESULTS: Homocysteine levels were elevated in all OSAS groups and were statistically significantly different between the mild and moderate/severe groups. Significant differences were present between the variables nocturnal oxygen desaturation (NOD), respiratory arousal and light sleep among the mild and moderate/severe groups. We found a significant positive correlation between homocysteine levels and NOD duration, and hsCRP levels were positively correlated with the apnea-hypopnea index and NOD duration. CONCLUSIONS: In all OSAS groups, homocysteine levels were elevated regardless of the presence of cardiac dysfunction. Echocardiographic abnormalities were primarily left-ventricular (LV) hypertrophy and LV diastolic dysfunction and could be observed in all OSAS severity groups.

Relationship between bone mineral density, bone turnover markers and homocysteine, folate and vitamin B12 levels in postmenopausal women.

PURPOSE: We aimed to investigate whether levels of homocysteine (Hcy), folate, and vitamin B12 are related to bone turnover markers and bone mineral density (BMD) in postmenopausal women. METHODS: One hundred and twenty postmenopausal women were divided into three groups: osteoporotic, osteopenic and normal, according to the BMD measurements. The age, weight, body mass index (BMI), years since menopause (YSM), gravidity, parity, bone turnover markers [type I collagen C-telopeptides (CTx) and bone-specific alkaline phosphatase (BAP)], serum Hcy, parathyroid hormone (PTH), vitamin B12, folate, calcium and magnesium levels were compared with each other. RESULTS: Twenty-five women had osteoporotic, 42 women had osteopenic, and 53 had normal BMD values. After adjusting for confounding factors, serum Hcy levels were significantly higher in osteoporotic women [adj OR = 38.95 (1.474-1029.88) p = 0.02]. The age, YSM, PTH, CTx and BAP levels were related to serum Hcy in all women (beta = 0.523, p = 0.0001; beta = 0.446, p = 0.001; beta = 0.295, p = 0.005; beta = 0.239, p = 0.026; beta = 0.451, p = 0.001, respectively). CONCLUSIONS: Our data showed that vitamin B12, folate and Hcy levels were not related with BMD in postmenopausal women. We think that one of the underlying mechanisms of increased Hcy levels and osteoporosis may be a mechanistic link which cannot detected by BMD or biochemical markers.

Institutional experience of newborn screening for inborn metabolism disorders by tandem MS in the Turkish population.

Background The tandem mass spectrometry method in the screening of congenital metabolic disorders is not included in routine national newborn screening programmes in Turkey. To evaluate the distribution of acylcarnitines and amino acid levels in normal newborns, establish acylcarnitine and amino acid cut-off levels and further preliminary results of inherited metabolic disorders inferentially in the Turkish population. Methods Newborn screening tests performed by tandem MS from 2016 to 2018 were retrospectively reviewed. The study group included 17,066 newborns born in our hospitals located in various regions of Turkey. Blood samples were obtained from infants older than 24 h of age. Among the 17,066 newborns, the metabolic screening data of 9,994 full-term newborns (>37 weeks) were employed to obtain the percentile distribution of the normal population. The study group (17,066) was screened for 26 types of inborn error of metabolism. Results Our established cut-offs, were compared with the cut-offs determined by Region for Stork Study and Centers for Disease Control. Among the 26 screened disorders, a total of 12 cases (8 amino acid metabolism disorders, 1 urea cycle defect, 2 organic acidaemias and 1 fatty acid oxidation disorder) were identified. Conclusions Because of the high rate of consanguineous marriages in Turkey, the development of a nationwide screening panel is necessary for early detection and management of potentially treatable inherited metabolic disorders.

An Assessment of HbA1c in Diabetes Mellitus and Pre-diabetes Diagnosis: a Multi-centered Data Mining Study.

HbA1c test has been widely used to evaluate glycemic control in patients with diabetes. However, there are controversial results regarding the value of HbA1c in the diagnosis of diabetes mellitus (DM). The present study investigates the diagnostic effectiveness of HbA1c in a large patient group. The oral glucose tolerance test and HbA1c results of 6551 patients (4704 healthy, 1345 pre-diabetes, 502 DM) in 12 different medical centers in Turkey between 2010 and 2016 were examined to understand the effectiveness of HbA1c in the diagnosis of DM. Different Roche systems were used for measuring HbA1c via the immunoturbidimetric method. The DM ROC curves revealed the diagnostic sensitivity, specificity, and AUC of 74.5%, 87.1%, and 0.866 (CI 95% 0.858-0.875), respectively, for HbA1c (at the cut-off 41 mmol/mol, 5.9%). For HbA1c at the universal diagnostic decision value of 48 mmol/mol (6.5%), the sensitivity and specificity were determined as 32.4% and 99.9%, respectively. The ROC curves for fasting plasma glucose (FPG) revealed the diagnostic sensitivity, specificity, and AUC of 71.3%, 85.3%, and 0.853 (CI 95% 0.844-0.861), respectively. However, the ROC curve results for pre-diabetes (HbA1c at the cut-off value of 39 mmol/mol, 5.7%) revealed the diagnostic sensitivity, specificity, and AUC of 45.7%, 76.4%, and 0.641, respectively. Furthermore, it was shown that the changes in HbA1c values due to gender and age had no clinical effect on the diagnosis. According to our results, it remains challenging to suggest HbA1c measurements can have a significant contribution to the FPG measurements. It was found that the sensitivity is specifically low in the assessment of the pre-diabetes data. Additionally, considering the problems associated with Hb1Ac measurements, further studies conducted in different regions by using different methods are required.

The effect of vitamin C on ischemia reperfusion injury because of prolonged tourniquet application with reperfusion intervals.

We examined the effect of vitamin C on muscle injury distal to the tourniquet which was applied for 4 hours with 10- and 20-minute reperfusion intervals after 2 hours of tourniquet. Sixty-four Sprague-Dawley rats were allocated to 4 randomized groups. After 2 hours tourniquet, 10- and 20-minutes of reperfusion were allowed to half of each group respectively. Afterward an additional 2 hours compression was applied. Except the control group the animals received vitamin C intravenously, before the first tourniquet in Group I, at the reperfusion interval in Group II, and at both times in Group III. Malondialdehyde levels were measured in blood and the tibialis anterior muscle. The muscle was histopathologically examined. The data was evaluated statistically. The effects of timing and the dose of vitamin C on ischemia reperfusion injury remain controversial and there was no statistical difference between 10- and 20-minute reperfusion intervals. But the blood malondialdehyde levels showed that vitamin C has a positive effect on the muscle injury caused by ischemia-reperfusion.

Serotonin dilemma in postmenopausal women: is it low or high?

OBJECTIVE: In this study, we have studied with premenopausal (PM), naturally menopausal (NM) and surgically induced menopausal (SM) women in order to investigate the differences in serum cortisol, dehydroepiandrosterone sulfate (DHEA-S), follicle stimulating hormone (FSH) and estradiol (E2) levels on serum serotonin levels. METHODS: Forty premenopausal (36.7+/-3.5 years), 40 naturally menopausal (54.2+/-8.4 years) and 38 surgically induced menopausal (55.4+/-11.2 years) women were included in the study. None of the subjects were using antidepressants or hormone replacement therapy. In NM and SM, years since menopause (YSM) were 3.16+/-1.58 and 3.36+/-1.89, respectively. Cortisol, DHEA-S, FSH and E2 levels were determined by immunochemiluminisence while serotonin levels were determined by HPLC. RESULTS: Serum serotonin levels in NM women were higher than the other two groups [144.23+/-45.29 microg/L vs 61.35+/-37.72 microg/L in SM women and 98.74+/-50.29 microg/L in PM women]. E2 and DHEA-S were positively correlated, while FSH and cortisol were negatively correlated with serotonin in NM and SM. There was no significant correlation between serotonin and age or YSM. In the PM group, there was no significant correlation between serotonin and the hormones. CONCLUSION: In conclusion, increased serotonin levels in naturally menopausal women may be a compensatory mechanism to decreased E2 levels as it is postulated that there is strong interaction between E2 and the serotoninergic system.

Coagulation status and biochemical and inflammatory markers in multiple sclerosis.

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system and is the most common cause of neurologic disability in young adults. In this study, the coagulation status and biochemical and non-specific inflammatory markers in patients with MS were investigated. Plasma prothrombin time, activated partial thrombin time, fibrinogen, D-dimer, serum high sensitive C-reactive protein, homocysteine, blood urea nitrogen, creatinine, calcium, total protein, albumin, total cholesterol, vitamin B12, folate levels and erythrocyte sedimentation rate were measured in 42 patients with MS and 31 healthy subjects as a control group. There was a positive correlation between homocysteine and D-dimer levels (r=0.84, p<0.01). However, there was no significant correlation between homocysteine, vitamin B12 (r=0.18) and folate (r=0.23) levels. Serum total protein, albumin and calcium levels of MS patients were lower than the control group. There are some alterations in the coagulation and biochemical status in MS patients. These findings may contribute to better understanding of the etiopathogenesis and clinical characteristics of this disease.

Glutaric acidemia type II patient with thalassemia minor and novel electron transfer flavoprotein-A gene mutations: A case report and review of literature.

Glutaric acidemia type II (GAII), also known as multiple acyl-CoA dehydrogenase deficiency, is an autosomal recessive inborn error of amino acid and fatty acid metabolism. We report a case of GAII with novel electron transfer flavoprotein (ETF)-A mutations in a 2-year-old female with thalassemia minor. The patient developed an episode of hypoglycemia and hypotonicity on the postnatal first day. Laboratory investigations revealed elevations of multiple acyl carnitines indicating glutaric acidemia type II in newborn screening analysis. Urinary organic acids were evaluated for the confirmation and revealed a high glutaric acid excretion. Genetic analysis revealed two novel mutations in the ETF-A gene, which are considered to be compound heterozygote. At the 8 mo of life ketone therapy was added, which significantly increased the neuromotor development. The patient had been closely followed for two years with carnitine, riboflavin, coenzyme Q10, and ketone supplementation in addition to a high carbohydrate diet. Although the patient had comorbidity like thalassemia minor, her neuromotor development was normal for her age and had no major health problems. This specific case expands the previously reported spectrum of this disease.