Cost analysis of 25-hydroxy vitamin D tests in Turkiye with big data: A cross-sectional study.

OBJECTIVE: The high prevalence of vitamin D deficiency in the population causes physicians to request more vitamin D tests and increases laboratory costs. It is aimed at investigating the demanded numbers and cost analyzes of 25-hydroxyvitamin D (25(OH)D) tests with the big data obtained from the national information health system of the Turkish Ministry of Health. METHODS: Between 2017 and 2018, all inpatient and outpatient tests and 25(OH)D tests in all medical biochemistry laboratories in Turkiye were determined based on department and institution type. The cost amount, distribution among health institutions, and test request rates were calculated. In both years, the top ten most expensive tests, according to health institutions, were evaluated. RESULTS: The total number of medical biochemistry tests performed in 2017 and 2018 was 1.424.948.155 and 1.713.134.326, respectively. The number of 25 (OH)D tests analyzed in the same years was 8.698.393 and 13.919.127, respectively. When the data of the 2 years are compared, the consumption of 25 (OH)D tests increased by 37% in General hospital laboratories, whereas it increased by 115.09% in primary health laboratories. When all health institutions were evaluated, the increase rate in 25 (OH)D test demand was 60%, while the cost increase rate was 23%. CONCLUSION: This report showed that the demands for 25(OH)D testing are increasing steeply, especially in primary health-care facilities. In this direction, laboratory information system test demand restrictions in accordance with national and international guidelines are important issues for policymakers.

Investigation of the effect of vitamin K1 prophylaxis on newborn screenings tests in newborns.

Background: Routine screening for hereditary disorders in newborns includes screening for treatable metabolic and endocrine disorders, such as biotidinase deficiency, galactosemia, maple syrup urine disease, hypothyroidism, and cystic fibrosis. Incorrect test results may be encountered due to the use of vitamin K1. To investigate the interference effect of vitamin K1 on neonatal screening tests and to raise awareness of erroneous measurements. Methods: Heel blood samples were taken from 25 newborns born in a neonatal intensive care unit. Dry blood C0, C2, C3, C4, C4DC, C5:1, C5OH, C5DC, C6, C6DC, C8, C8:1, C8DC, C10, C10:1, C10DC, C12, C14, C14:1, C14:2, C16, C16:1, C18, C18:1, C18:2, C18:OH, methylglutaryl, valine, leucine/isoleucine, methionine, phenylalanine, argininosuccinic acid, aspartate, alanine, arginine, citrulline, glycine, ornithine, and glutamate tests were studied using the tandem mass spectrometry (MS) method. The results of the heel blood samples obtained before and after the application of vitamin K1 (Phyto menadione) were compared. Results: In two studies conducted with in vitro and in vivo tests, C0, C2, C3, C4, C4DC, C5, C5OH, C6, C8, C10, C10:1, C14, C16, C16:1, C18, C18:1, methylglutaryl, phenylalanine, argininosuccinic acid, tyrosine, aspartate, arginine, citrulline, glycine, and glutamine were all significantly elevated (p < 0.05). Conclusions: Heel blood samples may yield false results due to vitamin K1 administration. In the case of doubtful results, a new sample should be taken and the measurement should be repeated.

Relationship Between the Coma Scores and Hemogram Parameters Measured by New-generation Devices.

OBJECTIVE: To investigate the relationship between the coma scores-Glasgow coma scale (GCS), Sequential Organ Failure Assessment (SOFA), and Acute Physiological and Chronic Health Assessment (APACHE-II)-in intensive care unit (ICU) patients and the percentage of macrocytosis (%MAC), immature granulocyte (IG), cellular haemoglobin concentration (cHGB), nucleated red blood cell (NRBC), nucleated red cell/ white cell ratio (NR/W), hyperchromic ratio (%HPR), and platelet distribution width (PDW) values. STUDY DESIGN: A descriptive comparative study. Place and Duration of the Study: Medicine Faculty, University of Harran, Turkiye, from December 2020 to May 2022. METHODOLOGY: The hemogram parameters of the patient groups with a GCS of 3-8 (n=51) and a GCS of 9-15 (n=43) and a control group of 55 healthy volunteers were measured using the new-generation hemogram autoanalyzer AlinityHQ (Abbott, USA). These parameters were compared with the coma scores (GCS, SOFA, and APACHE-II) of the patients. RESULTS: There was a statistically significant difference in IG, %MAC and PDW values (p-values were 0.025, 0.011, and 0.004, respectively) and an inverse correlation with GCS scores (correlation coefficients were -0.247, -0.264, and -0.297, respectively) was observed. There was also a significant correlation between the SOFA scores and %HPR and cHGB (correlation coefficients were 0.234, -0.358, p-values were 0.025, 0.001, respectively), and the APACHE-II scores and NRBC and NR/W values (correlation coefficients were -0.270, -0.247, p values were 0.009, 0.017, respectively). CONCLUSION: While other haematological parameters other than PDW were not associated with coma scores, parameters measured using new-generation haematological devices (%MAC, IG, cHGB, NRBC, NR/W, and %HPR) were found to be associated with estimated coma scores. These parameters can therefore be used as simple, rapid prognostic biomarkers and assist researchers in the development of new scoring models. KEY WORDS: ICU, Hyper, Coma, Sofa, Apache.

Effects of COVID-19 pandemic lockdown on the metabolic control of type 2 diabetes mellitus in patients.

Objective: The effects of the COVID-19 pandemic on the control of diabetes mellitus in patients are largely unknown. In this study we aimed to analyze the impact of the pandemic and the ensuing lockdown on the management of type 2 diabetes mellitus. Subjects and methods: A total of 7,321patients with type 2 diabetes mellitus (4,501 from the pre-pandemic period, 2,820 from the post-pandemic period) were studied retrospectively. Results: The admission of patients with diabetes melitus (DM) decreased significantly during the pandemic (4,501 pre-pandemic vs. 2,820 post-pandemic; p < 0.001). The mean age of patients was statistically lower (51.5 ± 14.0 vs. 49.7 ± 14.5 years; p < 0.001), and the mean glycated hemoglobin (A1c) level was significantly higher (7.9% ± 2.4% vs. 7.3% ± 1.7%; p < 0.001) in the post-pandemic period than in the pre-pandemic. The female/male ratio was similar in both periods (59.9%/40.1% for pre-pandemic, 58.6%/41.4% for post-pandemic; p = 0.304). As calculated by month the pre-pandemic rate of women was higher only in January (53.1% vs. 60.6%, p = 0.02). Mean A1c levels were higher in the postpandemic period than in the same month of the previous year, excluding July and October (p = 0.001 for November, p < 0.001 for others). Postpandemic patients admitted to the outpatient clinic were significantly younger than prepandemic visits for July (p = 0.001), August (p < 0.001) and December (p < 0.001). Conclusion: The lockdown had detrimental effects on blood sugar management in patients with DM. Hence, diet and exercise programs should be adapted to home conditions, and social and psychological support should be provided to patients with DM.

False negative effect of high triglycerides concentration on vitamin D levels: A big data study.

Background: Inaccurate test results may be a reason why vitamin D deficiency is seen as a common problem worldwide. Interferences from the sample matrix during testing are the most important factors in measurement errors. In this study, the relationship between triglycerides and total cholesterol levels and vitamin D levels in Turkey was investigated. Methods: The 25-hydroxyvitamin D test results and lipid test results studied in Turkey in 2021 were compared. Data were obtained from the Ministry of Health National Health Database. Simultaneously, 25-hydroxyvitamin D, triglyceride, and total cholesterol levels were studied, and 1,135,644 test results were taken as the basis. Results: In the group of patients with total cholesterol levels between 0-10.33 mmol/L, the proportion of patients below 20 mg/L ranged from 56.8% to 61.8%. In the patient group with cholesterol between 10.36-259 mmol/L, the rate of patients with less than 20 mg/L was between 70.8-100%, while the rate of patients with cholesterol above 100 mg/L was 0%. The mean 25-hydroxyvitamin D level was 20.1 mg/L in the patient group with a total cholesterol level between 0-10.33 mmol/L, and 16 mg/L in the patient group with a cholesterol level above 10.36 mmol/L. The mean 25-hydroxyvitamin D level was 20.11 mg/L in the patient group with triglycerides 0-10.16 mmol/L, and the 25-hydroxyvitamin D level was 12.28 mg/L in the patient group with triglycerides 10.17-113 mmol/L. The proportion of patients with vitamin D levels above 100 mg/L was found to be 0% in the group of patients with triglycerides above 10.17-113 mmol/L. Conclusions: According to this study, there is a risk of toxicity when administering vitamin D therapy in patients with high cholesterol and triglycerides levels. This study is the first of this size in the literature. High triglycerides and cholesterol levels can cause inaccurate measurement of vitamin D levels, so care should be taken when evaluating these tests.

Determination of Serum Differential Carnitine Ester Levels in HIV(+) Patients: A Cross-Sectional Study.

OBJECTIVE: It has been reported that carnitine deficiency is observed in various viral infections and in the follow-up of the prognosis of some diseases. In this cross-sectional study, we aimed to determine how carnitine ester derivatives change in HIV-positive patients. MATERIALS AND METHODS: In this study, 25 HIV-infected patients who applied to Harran University Faculty of Medicine Education Research and Practice Hospital Infectious Diseases and Clinical Microbiology Outpatient Clinic and who did not receive any antiretroviral treatment, as well as 25 healthy volunteers were included in the study. Carnitine ester levels in serum samples were measured by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) method (Shimadzu North America, Columbia, MD, USA). RESULTS: While suberoylcarnitine (C8DC), myristoleylcarnitine (C14:1), tetradecadienoylcarnitine (C14:2), palmitoleylcarnitine (C16:1), and linoleylcarnitine (C18:2) levels in HIV(+) patients were quite low compared to the control group, tiglylcarnitine (C5:1) levels were high (p ≤ 0.05). In addition, C5:1 and C14:2 index parameters according to VIP score, and C5:1 and C14:1/C16 index parameters according to ROC analysis were determined as markers with high potential to distinguish HIV(+) patients from healthy volunteers. CONCLUSION: This study showed that levels of acylcarnitine derivatives might be altered in HIV(+) patients, and the results obtained may contribute to a better understanding of carnitine metabolism.

Effects of propofol and dexmedetomidine on carnitine metabolism in normal human bronchial epithelial cells.

Propofol and dexmedetomidine (DEX) are widely used for anesthesia and sedation. We investigated the effects of propofol and DEX separately and in combination on the metabolic profile of carnitine in cultured normal human bronchial epithelial cells (BEAS-2B). Cells of the propofol group were cultured with 2 µg/ml propofol in RPMI-1640 medium. Cells of the DEX group were cultured with 0.2 ng/m DEX in RPMI-1640 medium. Cells of the propofol + DEX group were cultured with 2 µg/ml propofol + 0.2 ng/ml DEX in RPMI-1640 medium. The control group was untreated. Cells were incubated for 3 h following treatments. The effects of the drugs on cell viability were assessed using the MTT method and by microscopic examination following staining with acridine orange/ethidium bromide. The effects of drugs on carnitine, acetyl carnitine and 25 acylcarnitine derivative profiles were analyzed using liquid chromatography-tandem mass spectrophotometry. Neither propofol nor DEX affected cell viability. Administration of propofol, DEX or propofol + DEX to BEAS-2B cells caused no significant change in the concentrations of carnitine and acylcarnitine derivatives compared to the control group. We found that propofol and DEX exhibit no negative effects on the carnitine metabolism by BEAS-2B cells in vitro at clinically relevant concentrations. Our findings establish a baseline for clinical studies of the effects of propofol and DEX on carnitine metabolism.

Effects of COVID-19 pandemic lockdown on the metabolic control of type 2 diabetes mellitus in patients

ABSTRACT Objective: The effects of the COVID-19 pandemic on the control of diabetes mellitus in patients are largely unknown. In this study we aimed to analyze the impact of the pandemic and the ensuing lockdown on the management of type 2 diabetes mellitus. Subjects and methods: A total of 7,321patients with type 2 diabetes mellitus (4,501 from the pre-pandemic period, 2,820 from the post-pandemic period) were studied retrospectively. Results: The admission of patients with diabetes melitus (DM) decreased significantly during the pandemic (4,501 pre-pandemic vs. 2,820 post-pandemic; p < 0.001). The mean age of patients was statistically lower (51.5 ± 14.0 vs. 49.7 ± 14.5 years; p < 0.001), and the mean glycated hemoglobin (A1c) level was significantly higher (7.9% ± 2.4% vs. 7.3% ± 1.7%; p < 0.001) in the post-pandemic period than in the pre-pandemic. The female/male ratio was similar in both periods (59.9%/40.1% for pre-pandemic, 58.6%/41.4% for post-pandemic; p = 0.304). As calculated by month the pre-pandemic rate of women was higher only in January (53.1% vs. 60.6%, p = 0.02). Mean A1c levels were higher in the postpandemic period than in the same month of the previous year, excluding July and October (p = 0.001 for November, p < 0.001 for others). Postpandemic patients admitted to the outpatient clinic were significantly younger than prepandemic visits for July (p = 0.001), August (p < 0.001) and December (p < 0.001). Conclusion: The lockdown had detrimental effects on blood sugar management in patients with DM. Hence, diet and exercise programs should be adapted to home conditions, and social and psychological support should be provided to patients with DM.

25-hydroxyvitamin D (25-OHD) levels in Turkish geriatric population: A nationwide study.

Background: Across the world, 25-hydroxyvitamin D (25-OHD) deficiency is a major health problem associated with many chronic diseases in the geriatric population. Prior to this study, there were no data regarding 25-OHD levels among individuals over the age of 65 in Turkey. The aim of this study was to assess 25-OHD levels and seasonal variations in these values among people over the age of 65 in Turkey. Methods: This study included vitamin D measurements taken in 2016, 2017, and 2018 from the Turkish population over the age of 65. The age, gender, and seasonal average data of the study population were defined. The study data were obtained from the database of the Ministry of Health, and a Kolmogorov-Smirnov test was used to assess the distribution of the data. Medians and interquartile ranges (IQRs) were calculated for all categories, as the data were nonparametric. Results: The number of vitamin D measurements taken from the geriatric individuals included in this study was 305,329 for 2016, 576,452 for 2017, and 752,837 for 2018. The medians and IQRs of the 25-OHD levels in this population were 16 µg/L (IQR 7.45-24.55 µg/L) for 2016, 16.1 µg/L (IQR 7.8-24.4 µg/L) for 2017, and 16.4 µg/L (IQR 8.95-23.85 µg/L) for 2018. Conclusions: While the 25-OHD levels of older men tended to increase during the period of seasonal sunlight in Turkey, this variability was observed in elderly women. This suggests that older women tend to live more sedentary lives and have insufficient sun exposure. Overall, the median 25-OHD levels of individuals over the age of 65 tended to decrease each year.

Plasma Free Amino Acid Profile in HIV-Positive Cases.

BACKGROUND: Increasing the sensitivity and availability of liquid chromatography tandem mass spectrometry (LC-MS/MS) devices may provide advantages in terms of revealing the changes in metabolic pathways in HIV-positive patients and elucidating the physiopathology. INTRODUCTION: The aim of this study was to determine the difference in amino acid levels between HIV-positive patients and healthy individuals by using LC-MS / MS and investigate its relationship with HIV infection. MATERIAL AND METHODS: Concentrations of 36 different amino acids and their derivatives were measured and compared in venous plasma samples from 24 HIV-positive patients and 24 healthy individuals by using the LC-MS/MS method (Shimadzu North America, Columbia, MD, USA). RESULTS: HIV-positive subjects had significantly lower alanine, 1-methyl-L-histidine, valine, aspartate, cysteine, cystine, methionine, lysine, glutamine, imino acid, tyrosine, tryptophan, threonine, sarcosine, and argininosuccinic acid and significantly higher 3-methyl-L -histidine, asparagine, glutamate, and carnosine levels as compared to healthy controls. No significant differences were detected in other amino acids. CONCLUSION: The significant differences in amino acid profile between HIV-positive and healthy subjects may represent an auxiliary biomarker of cellular damage in asymptomatic HIV-positive patients that may be examined in more detail in further studies. It may also provide guidance for symptomatic cases in terms of the association between symptoms, clinical manifestations, and deficiency or excess of certain amino acids in the context of the complete metabolomics record of HIVpositive patients.

Do inflammatory markers such as NLRP3 and IL1-ß have an effect on the prediction of testicular sperm extraction success in infertile men?

Infertility is defined as the inability of couples to have a baby without contraception after at least 1 year of regular sexual intercourse. Mechanisms to explain inflammation in male infertility of unknown causes are still being investigated. The inflammasome is a key regulator of innate immunity, which is involved in the inflammatory response to infections and various diseases through the activation of caspase-1 and the use of inflammatory cytokines. Although many factors are believed to affect the success of mTESE in infertile patients with non-obstructive azoospermia (NOA), the inflammation mechanisms in the environment have not been clearly explained in the aetiology of infertility. In this study, we aimed to investigate the effect of NLRP3 and similar inflammasome mechanisms on antioxidant mechanisms on the success of mTESE. A total of 24 NOA patients with micro-testicular sperm extraction (mTESE +) and no sperm found (mTESE -) participated in the study between January 2020 and January 2021. NLRP3, IL1-ß, TAS, TOS and OSI amounts in serum and seminal plasma parameters were compared statistically, and their effects on mTESE success were investigated. FSH, LH, estradiol and testosterone values did not differ significantly (p > 0.05) in the group with mTESE (-) and mTESE (+). Serum IL-1 Beta, NLRP3, TOS, TAS, and OSI values did not differ significantly (p > 0.05) in the group with mTESE (-) and mTESE (+). Seminal plasma TOS and OSI values were significantly lower in the group with mTESE (+) than the group with mTESE (-). Although inflammasomes such as NLRP3 and IL1-ß do not have a significant predictive value in the success of mTESE, we think the high seminal plasma values of infertile patients may be understandable with further studies. This study was conducted to determine how inflammasomes are involved in IL-1ß, pathway NLRP3, and sperm retrieval in micro testicular sperm extraction (mTESE) in infertile men.

Transcutaneous Electrical Nerve Stimulation (TENS) for the Treatment of Renal Colic in the ED: A Randomized, Double-Blind, Placebo-Controlled Trial.

STUDY OBJECTIVE: To determine the analgesic efficacy of TENS treatment in patients with renal colic in the emergency department (ED). METHODS: This double-blind, randomized controlled trial was conducted in a tertiary care ED. Patients with a definitive diagnosis of renal colic were assigned (1:1) as randomized to receive the real TENS with frequency 100 Hz, pulse width 200 microseconds, voltage 2 mA, or placebo with sham TENS. Pain intensity was measured using visual analog scales (VAS) at baseline, after 15 and 30th minutes. RESULTS: A total of 100 patients were included in the final analysis: 50 patients treated with real TENS and 50 patients treated with sham TENS. VAS scores in both groups were similar at baseline. The mean reduction in VAS score at 15 min was 33.3 ± 17.6 (95% Confidence interval (CI): 28.3 to 38.3) for the real TENS group and 14.9 ± 11.6 (95% CI 11.6 to 18.2) for the sham TENS group (mean difference: 18.4 (95% CI: 12.5 to 24.4, P < 0.0001). The mean reduction in VAS score at 30 min was 63.7 ± 21.1 (95% CI: 57.7 to 69.7) for the real TENS group and 14.9 ± 16.2 (95% CI: 19.5 to 10.3) for the sham TENS group (mean difference: 48.8, 95% CI: 41.4 to 56.3, P < 0.0001). Four patients (8%) in the real TENS group and 24 patients (48%) in the sham TENS group required the rescue medication after 30th minutes. CONCLUSIONS: TENS is effective for acute pain treatment in renal colic patients in the ED. TENS therapy could be a treatment option for renal colic.

Evaluation of oxidative stress in degenerative rotator cuff tears.

BACKGROUND: Oxidative stress occurs as a result of the disruption of the balance between the formations of reactive oxygen species and antioxidant defense mechanisms during the conversion of nutrients into energy. Increased body oxidative stress has been reported to be involved in the etiology of several degenerative and chronic diseases. We hypothesized that the body oxidative stress level is higher in patients with atraumatic degenerative rotator cuff tear than that in healthy individuals. METHODS: The patients who underwent arthroscopic repair for atraumatic, degenerative rotator cuff tear were prospectively evaluated. A total of 30 patients (group 1, 19 females and 11 males; mean age: 57.33 ± 6.96 years; range: 50-77 years) and 30 healthy individuals (group 2, 18 females and 12 males; mean age: 56.77 ± 6 years; range: 51-72 years) were included in the study. The Constant and American Shoulder and Elbow Surgeons scoring systems were used to evaluate the clinical outcomes. Serum oxidative stress parameters of the patients and the control group were biochemically evaluated. Accordingly, thiol/disulfide (DS) balance (DS/native thiol [NT], DS/total thiol [TT]), Total Oxidant Status (TOS), oxidative stress index, and nuclear factor erythroid-2-associated factor-2 values were used as the biochemical parameters indicating an increase in the serum oxidative stress level. Total antioxidant status and NT/TT values served as the biochemical parameters indicating a decrease in the serum oxidative stress level. RESULTS: The study follow-up duration was 12 months. A statistically significant increase was observed in American Shoulder and Elbow Surgeons and Constant scores of patients who underwent arthroscopic rotator cuff repair relative to that during the preoperative period (P = .01). The values of biochemical parameters (DS/NT, DS/TT, TOS, oxidative stress index, and nuclear factor erythroid-2-associated factor-2), which indicated an increase in the serum oxidative stress, were significantly higher in preoperative patients than those in postoperative patients, albeit the control group values were significantly lower than those of the postoperative patients. The biochemical parameters (NT/TT and total antioxidant status) indicating a decrease in the serum oxidative stress levels were significantly higher in the postoperative patients than those in the preoperative patients and significantly lower than those in the control group. CONCLUSION: High levels of markers indicating an increase in the serum oxidative stress in patients with degenerative rotator cuff rupture suggested that TOS may be involved in the etiopathogenesis of rotator cuff degeneration. Although the oxidative load decreases during the postoperative period, the fact that it is still higher than that in healthy individuals supports this claim.

The Relationship Between the Severity of Coronary Artery Disease and Erythrocyte Morphology Parameters Measured by New-Generation Hematology Analyzer.

BACKGROUND: There is a possible relation between red blood cell distribution width (RDW) and various clinical conditions. These conditions can render RDW disadvantageous in its relation with cardiovascular disease. There may be a relation between the severity of acute coronary syndrome (ACS) and the percentage of hypochromia (hypo%), percentage of hyperchromia (hyper%), percentage of macrocytosis (MAC%), and percentage of microcytosis (MIC%) values measured using new-generation hematological devices. OBJECTIVE: We aimed to examine the relation between the SYNTAX score and the hypo%, hyper%, MAC%, and MIC% values in patients admitted with ACS. METHODS: A group of 55 patients who underwent coronary angiography with diagnosis of ACS (STEMI and NSTEMI) and a control group of 48 patients with normal coronary arteries were included in the study. Venous blood samples were collected in the morning after a fasting of at least 8 h and analyzed using standard laboratory methods. Hemogram parameters were studied using Alinity HQ (Abbott, USA) a completely automated hemogram autoanalyzer. Biochemical parameters were studied using Architect c16000 (Abbott, USA) a completely automated biochemistry autoanalyzer. RESULTS: Significant difference was observed in erythrocyte morphology-related tests (mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, RDW, hypo%, hyper%, MIC%, and MAC%) between the groups. Correlation analysis showed a positive correlation between the SYNTAX score and MAC% (r = 0.315, p = 0.019). Multivariate logistic regression analysis was performed for MAC% to identify the independent predictors of the SYNTAX score (ß = 0.315, p = 0.019). CONCLUSION: Changes in MAC% test can be measured in emergencies with new-generation hematological devices and used as independent predictors of the presence of severe coronary artery disease.

A Pilot Study for Investigation of Plasma Amino Acid Profile in Neurofibromatosis Type 1 Patients.

BACKGROUND: Neurofibromatosis, also known as Von Recklinghausen disease, is a systemic and progressive genetic disease that primarily affects the skin, eyes, nervous system, and bones. The disease can occur in a variety of ways and can vary in individuals. Metabolomic-based research using blood samples has enabled new diagnostic methods to be used in the diagnosis of various diseases, especially cancer. Among the metabolites, profiling of plasma free amino acids (PFAA) is a promising approach because PFAAs bind all organ systems and play an important role in the metabolism. OBJECTIVE: This study aimed to determine the characteristics of PFAA profiles in neurofibromatosis patients and the possibility of using them for early detection and treatment of the disease. METHODS: Patients with a diagnosis of Neurofibromatosis Type I confirmed by genetic analysis and healthy individuals of the same age group without any disease were included in the study. We analysed the nineteen plasma free amino acids (phenylalanine, proline, threonine, arginine, asparagine, cystine, valine, glutamate, tyrosine, serine, glutamine, glycine, tryptophane, leucine, lysine, methionine, isoleucine, aspartate and alanine) from neurofibromatosis Type I patients and control group by liquid chromatography tandem mass spectrometry (LC-MS/MS) in Metabolism Laboratory of Harran University Research and Application Hospital. The results of the plasma free amino acid levels were divided into 3 groups as essential, semi-essential, and non-essential. The differences in amino acid levels between groups were determined. RESULTS: The levels of eight amino acids (methionine, arginine, cystine, glutamine, proline, asparagine, serine, aspartate) were significantly altered in patients with neurofibromatosis type 1. In essential amino acids, methionine levels were significantly higher in the patient group than control group. While the levels of arginine and glutamine in semi-essential amino acids were statistically significantly higher in the patient group, a significant decrease was observed in cystine and proline levels compared to the control group's amino acid levels. In the non-essential amino acids group, asparagine, serine and aspartate amino acid levels were significantly higher in the patient group compared to the control group. CONCLUSION: The current research predicates that eight amino acids, namely methionine, arginine, cystine, glutamine, proline, asparagine, serine, aspartate can be considered to be valuable biomarkers for neurofibromatosis type I. This present study is the first to build models for neurofibromatosis Type I screening using plasma free amino acids and the amino acid profile will be able to guide the prediction of the complications that may occur during the course of the disease.

The Association between Parameters of Erythrocytes Morphology and Thrombophilia-related Mutations.

BACKGROUND: Alterations in erythrocyte morphology parameters have been identified and associated with hematological disorders and other chronic and cardiovascular diseases. Erythrocytes are abundant in thrombus content. Their hemoglobin density and differences in the ratio of macrocytic and microcytic cells may be associated with hypercoagulopathy in those with a history of thrombosis. OBJECTIVE: This cross-sectional study aimed to investigate the relationship between hemogram parameters and thrombophilia genetic parameters. METHODS: A total of 55 patients whose thrombophilia panel was reviewed due to the diagnosis of thrombosis were included in the study. % MIC, % MAC, % HPO, % HPR and all hemogram parameters were measured using Abbott Alinity HQ. Prothrombin G20210A, MTHFR C677T, MTHFR A1298C, Factor V Leiden G169A and PAI-1 4G/5G mutations were studied using Real Time- PCR. RESULTS: The MTHFR C677T mutation was detected in 58.2% of the patients. The Factor V Leiden mutation was detected in 5.5% of the patients. The MTHFR A1298C mutation was detected in 58.2%, The PAI mutation was detected in 74.5%, and the Factor 13 mutation was detected in 29% of the patients. Prothrombin G20210A mutation was not detected in any of the patients. Red blood cell (RBC) and hematocrit (Hct) values were higher in Factor 13 mutant group; the Hgb and Htc values were higher in the MTHFR C677T mutant group. The plateletcrit (PCT) and platelet (PLT) values were lower in MTHFR C677T mutant group. CONCLUSION: The MTHFR C677T and Factor 13 mutations may be associated with high Hct and Hgb, RBC, Hgb, and Htc values, respectively and coagulation tendency in patients with a history of thrombosis.

Inhibition of Interleukin-6 signaling: A novel therapeutic approach to necrotizing enterocolitis.

OBJECTIVES: This study investigated the effects of tocilizumab on the prevention and treatment of experimental necrotizing enterocolitis (NEC) in newborn rats. METHODS: Forty-two newborn Sprague-Dawley rats were randomly separated into three groups: NEC + placebo, NEC + tocilizumab, and the control group. NEC + placebo and NEC + tocilizumab groups were given 1 mg/kg lipopolysaccharide intraperitoneally once only on the first day, were fed with a special rodent formula every 3 h, inhaled 100% CO2 for 10 min, were exposed to cold stress at + 4 °C for 5 min, and 97% O2 for 5 min twice a day for 3 days. NEC + tocilizumab group was treated with 8 mg/kg/day tocilizumab (Actemra®) intraperitoneally, while NEC + placebo group was given intraperitoneal 0.9% saline at a dose of 2 mL/kg/day from the first day to the end of the study. All newborn rats were sacrificed on day 4. Specimens were taken for histopathologic, immunohistochemical and biochemical evaluation from the ileum and proximal colon. RESULTS: NEC + tocilizumab group had higher weight gain and survival rate compared to NEC + placebo group and clinical sickness score was reduced in NEC + tocilizumab group (p < 0.05). Lower tissue damage and apoptosis were found in the NEC + tocilizumab group compared to the NEC + placebo group (p < 0.01). Tissue Interleukin-6, Interleukin-1ß, TNF-α, myeloperoxidase and caspase-3 levels were significantly decreased in the NEC + tocilizumab group (p < 0.01). CONCLUSIONS: Tocilizumab could be a potential option in the prevention and treatment of NEC.

BTD Gene Mutations in Biotinidase Deficiency: Genotype-Phenotype Correlation.

OBJECTIVE: To identify the biotinidase (BTD) gene mutations in patients with biotinidase deficiency in our region; and to determine the phenotype-genotype correlations in the presence of clinical findings. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Medical Genetics and Pediatric Metabolism Outpatient Clinic, Faculty of Medicine, Harran University, between January 2018 and June 2020. METHODOLOGY: Two hundred and nine patients, who were found positive for biotinidase deficiency in heel blood screening, were included. Genomic DNA was isolated from peripheral blood. Next-generation DNA sequencing analysis was performed using primers covering the exon regions of the BTD gene. The results were analysed by the mutation surveyor programme. RESULTS: The most common mutation was c.1330 G>C (p.D444H) and the second most common mutation was c.470 G>A (p.R157H). The majority of the mutations are missense; and they are especially located in the exon 4. The most frequent mutations were found to be D444H and R157H with a rate of 66.66% in symptomatic patients. CONCLUSION: Common mutations in BTD deficiencies were indentified. Associating them with phenotype-genotype data will assist clinicians in better genetic counselling and management in the future by implementing prevention programmes. Key Words: Biotinidase deficiency, BTD gene, Newborn screening, Inherited metabolic disease, Newborn screening programme.

Inhibition of Carbonic Anhydrase IX Promotes Apoptosis through Intracellular pH Level Alterations in Cervical Cancer Cells.

Carbonic anhydrase IX (CAIX) is a hypoxia-related protein that plays a role in proliferation in solid tumours. However, how CAIX increases proliferation and metastasis in solid tumours is unclear. The objective of this study was to investigate how a synthetic CAIX inhibitor triggers apoptosis in the HeLa cell line. The intracellular effects of CAIX inhibition were determined with AO/EB, AnnexinV-PI, and γ-H2AX staining; measurements of intracellular pH (pHi), reactive oxygen species (ROS), and mitochondrial membrane potential (MMP); and analyses of cell cycle, apoptotic, and autophagic modulator gene expression (Bax, Bcl-2, caspase-3, caspase-8, caspase-9, caspase-12, Beclin, and LC3), caspase protein level (pro-caspase 3 and cleaved caspase-3, -8, -9), cleaved PARP activation, and CAIX protein level. Sulphonamide CAIX inhibitor E showed the lowest IC50 and the highest selectivity index in CAIX-positive HeLa cells. CAIX inhibition changed the morphology of HeLa cells and increased the ratio of apoptotic cells, dramatically disturbing the homeostasis of intracellular pHi, MMP and ROS levels. All these phenomena consequent to CA IX inhibition triggered apoptosis and autophagy in HeLa cells. Taken together, these results further endorse the previous findings that CAIX inhibitors represent an important therapeutic strategy, which is worth pursuing in different cancer types, considering that presently only one sulphonamide inhibitor, SLC-0111, has arrived in Phase Ib/II clinical trials as an antitumour/antimetastatic drug.