GLIADOMORPHIN AND CASOMORPHIN LEVELS IN CHILDREN WITH AUTISM SPECTRUM DISORDERS

High frequency of detection of various disorders of the gastrointestinal tract in patients with autism spectrum disorders (ASD) formed the basis of the first attempts to prescribe diet therapy. Despite the fact that diets are not officially included in the complex of therapeutic measures for ASD, the use of gluten-free (GFD) and casein-free diets (CFD) is widespread practice. The aim of the study was to assess the level of gliadomorphins (GM) and casomorphins (CM) in the blood of children with ASD, depending on the eating style and in comparison with healthy peers. Materials and methods of research: a prospective continuous open controlled study with the participation of 230 children aged 3–15 years. The first group included 36 patients with ASD who had been observing GFD for more than 6 months; in addition, 18 children were also on CFD. Boys in the group ware 23 (64%), girls – 13 (36%). The second group included 49 children with ASD who had no dietary restrictions. Boys – 35 (71%), girls – 14 (29%). The third (control) group consisted of 145 apparently healthy children: boys – 83 (57.3%), girls – 62 (42.7%). Determination of CM and GM levels was performed on a Multiskan Go analyzer (Thermo Fisher Scientific, Finland) by enzyme-linked immunosorbent assay using Human Gliadomorphin ELISA Kit and Human Casomorphin ELISA Kit (Hycult Biotech, Netherlands). Results: more boys than girls were included in all study groups; no statistically significant differences in gender distribution were found (p=0.2). The median age values in the 1st group was 6.5 [4.0; 8.0] (3; 15) years, in the 2nd – 6.0 [4.0; 7.0] (3; 15) years, in the 3rd – 8.0 [6.0; 11.0] (3; 15) years, the differences are statistically significant (p=0.0001). The content of GM in the blood serum of children with ASD, adhering to GFD, is statistically significantly lower by 2.6 times compared with children not adhering to diet therapy – 0.61 [0.41; 0.80] (0.12; 6.42) and 1.6 [0.70; 2.38] (0.18; 4.03) ng/ml (p<0.001) and does not differ statistically significantly from the level in the control group – 0.41 ng/ml [0.31; 0.72] (0.14; 4.18) ng/ml (p>0.005). The content of CM in the blood serum of children with ASD, adhering to CFD and GFD, is also 1.25 times lower than in children not adhering to diet therapy 1.85 [1.04; 2.07] (0.16; 3.83) and 2.34 [2.11; 2.72] (0.93; 3.29) pg/ml (p<0.001) and 1.3 times lower than the level of the control group – 2.45 [2.13; 2.89] (1.22; 26.88) pg/ml (p<0.001). Conclusion: the results obtained confirm the theory/possibility of exorphin intoxication of ASD development, which may be pathogenetically significant for some patients with ASD, which must be taken into account when implementing an individual approach to prescribing GFD, CFD.

Circulating Expressions of Gonadotropin Releasing Hormones and Risk of Ovarian Cancer

Background: Ovarian cancer (OC) is a worst type of malignancy in the field of gynecology. This is because ovarian tumors diagnosed at advanced stage of disease. The exact mechanism for its development is still unknown. Aim: The aim of this study is to measure the levels of steroidal hormones and their function in ovarian cancer progression. Methods: In the present study, fifty ovarian cancer patients and fifty control individuals were taken and serum was separated from their blood samples. The levels of steroid hormones were measured by ELISA kit methods. Results: Result of the current study determined the levels of E2, progesterone, testosterone, FSH, LH, 17-β-HSD-I, 17-β-HSD-II, cortisol and aromatase were extensively higher in patient group in comparison with healthy individuals. Conclusion: Current study concluded the Study concluded that overexpression of steroid hormones may lead to enhance tumor survival in ovarian cancer through various signaling mechanisms. Keywords: Ovarian cancer, Estradiol, FSH, LH, progesterone

Melatonin differences between day and night milk in primiparous holstein friesian and jersey dairy cattle

This study was conducted to determine the levels of melatonin in the day and night milk of Holstein and Jersey cows. In the study, samples of daytime milk produced from 27 head of Holstein and 27 head of Jersey cows in the first lactation, which were raised in a private dairy cattle enterprise in the Kaşınhanı neighborhood of Meram district of Konya city Turkey, and night milk samples taken from the same cows that were blackened for one (1) week were used. Melatonin levels in milk samples taken from day and night milk were determined separately for Holstein and Jersey cows with the help of Bovine Melatonin (MLT) Elisa Kit. In the study, it was determined that the ratio of melatonin in day and night milk in Holstein cows was 2.912 pg/ml and 11.314 pg/ml, respectively, and the ratio of melatonin in Jersey cows was 2.924 pg/ml and 6.954 pg/ml in the same order. The difference between the melatonin levels of the day and night milk of Holstein and Jersey cows was found to be statistically significant (p<0.01). At the end of the study, it can be stated that night milk can be used for medical purposes and a new production source may arise for producers since there is a significant difference in melatonin between day and night milk.

Increased Serum Levels of S100A4 and S100A15 in Individuals Suffering from Hidradenitis Suppurativa

Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease. Recently, some S100 proteins have been suggested to play an important role in the pathogenesis of chronic immune-mediated inflammatory diseases and they may constitute valuable biomarkers for these diseases’ diagnosis and monitoring. The objective of the current study was to investigate, for the first time, serum levels of S100A4 and S100A15 in individuals suffering from HS. Furthermore, we assessed the associations between S100A4 and S100A15 serum levels and the severity of disease, CRP serum concentration and some demographic and clinical data. Serum levels of S100A4 and S100A15 were evaluated with the commercially available ELISA kit according to the manufacturer’s instructions. The serum level of S100A4 in individuals with HS was significantly elevated as compared to controls, with the highest level found in the individuals in Hurley stage II. The S100A15 serum level was positively correlated with the CRP concentration and was associated with the severity of the disease. The serum level of S100A15 in the individuals in Hurley stage III was significantly elevated compared to that of the controls and the individuals with HS in Hurley stages I and II. S100A4 and S100A15 may be considered as new serum biomarkers for the monitoring of HS progression, and they may play a role in the pathogenesis of HS by promoting inflammatory process and fibrosis.

Gliadin proteins from wheat flour: the optimal determination conditions by ELISA

Introduction. The number of people with celiac disease is rapidly increasing. Gluten, is one of the most common food allergens, consists of two fractions: gliadins and glutenins. The research objective was to determine the optimal conditions for estimating gliadins by using enzyme-linked immunosorbent assay (ELISA). Study objects and methods. The experiment involved wheat flour samples (0.10; 0.20, 0.25, 0.50, and 1.0 g) suspended in different solvents (ethanol, methanol, 1-propanol, and isopropanol) of different concentrations (40, 50, 60, 70, 80, and 90% v/v). The samples were diluted with Tris buffer in ratios of 1:50, 1:100, 1:150, and 1:200. The gliadin test was performed using a Gliadin/Gluten Biotech commercial ELISA kit (Immunolab). Results and discussion. The optimal conditions for determining gliadin proteins that provided the highest gliadin concentration were: solvent – 70% v/v ethanol, extract:Tris buffer ratio – 1:50, and sample weight – 1.0 g. Conclusion. The obtained results can be of great importance to determine gliadin/gluten concentrations in food products by rapid analysis methods.

Correlation of Pentosidine with Kidney Diseases in Iraqi Patients with Diabetic Nephropathy

Diabetic nephropathy (DN) is a principle cause of microangiopathy and the main reason for kidney disease at the end stage in patients with type 2 diabetes mellitus (T2DM). This work aimed to study the relation of pentosidine with kidney injury in the case of diabetic nephropathy. This study included 75 patients suffering from T2DM and 75 apparently healthy subjects. The patients group was divided into three groups ((normoalbumin, microalbuminuria, and macroalbuminuria; 25 patients for each) on the basis of albumin-creatinine ratio (ACR) . The level of serum pentosidine was determined using an ELISA kit. The level of pentosidine was found to be significantly higher in DN patients than in the healthy group. Also, the results revealed a strong positive correlation of pentosidine with each of creatinine and blood urea levels, while a negative correlation was recorded with eGFR. It can be concluded that pentosidine may be associated with disease progression and it may be employed as one of the most efficient markers for the prediction of renal function.

Korelasi antara Neutrophil-Lymphocyte Ratio dan NT-proBNP pada Pasien Gagal Jantung Anak Akibat Penyakit Jantung Rematik

Latar belakang. Penyakit jantung rematik (PJR) merupakan penyebab paling sering gagal jantung anak didapat terutama di negara berkembang. Pemeriksaan penanda gagal jantung seperti N-terminal pro brain natriuretic peptide (NT-proBNP) dapat membantu dalam melakukan diagnosis. Neutrophil-lymphocyte ratio (NLR) merupakan penanda inflamasi yang mudah dilakukan dan sudah digunakan dalam berbagai penyakit kronis.Tujuan. Mengetahui korelasi antara nilai NLR dengan kadar NT-proBNP pada pasien gagal jantung anak akibat PJR.Metode. Penelitian analitik observasional dengan studi potong lintang. Data berasal dari register gagal jantung anak akibat penyakit jantung rematik Divisi Kardiologi Departemen/KSM Ilmu Kesehatan Anak RSUP Dr. Hasan Sadikin Bandung Tahun 2020 dan sampel BBT penelitian sebelumnya. Populasi penelitian ini adalah anak berusia <18 tahun yang memenuhi kriteria inklusi. Kadar NT-proBNP diperiksa menggunakan ELISA Kit Elabscience Catalog No: E-EL-H0902. Analisis korelasi dilakukan dengan menggunakan uji korelasi Spearman dengan aplikasi SPSS 22.Hasil. Total subjek penelitian 34 anak dengan usia median 14 tahun. Subjek penelitian paling banyak terdapat pada kelompok usia 11-<16 tahun (65%). Sebanyak 23 (67%) subjek berada pada derajat gagal jantung ringan berdasarkan klasifikasi NYHA kelas I-II. Keterlibatan katup paling banyak mencakup kerusakan katup mitral dan aorta. Peningkatan kadar NT-proBNP terdapat pada 18 subjek (53%). Rata-rata nilai NT-proBNP adalah 388,09 pg/ml (20-2500 pg/ml) dan NLR 1,79 (0,57-5,27). Hasil analisis korelasi antara NLR dan NT-proBNP menunjukkan tidak terdapat korelasi dengan nilai r=0,054 (p=0,382).Kesimpulan. Penelitian ini tidak menemukan korelasi antara nilai NLR dan kadar NT-proBNP pada pasien gagal jantung anak akibat PJR.

Quantification of Sphingosine-1-Phosphate (S1P) in Patients with Periodontitis Stage II Grade B

The aim of this study was the quantification of Sphingosine-1-phosphate (S1P) in periodontal pockets of patients with periodontitis. This is an observational, descriptive, case-control study. Thirty subjects were selected: 15 controls and 15 cases. A periodontal study was conducted following the parameters of AAP 2017 for the diagnosis of periodontal diseases. A sample of saliva and gingival crevicular fluid was obtained from each subject and then analyzed with the Human S1P Elisa kit (MyBioSource #MBS2516132) accordingly to the manufacturer's instructions, in order to verify the presence of S1P and quantify it´s concentration when founded. Results showed a significant difference (p=0.05) between cases and controls. In the case of saliva samples, the concentration of S1P was higher than the ones found in the control group (72.94 ng/mL and 45.12 ng/mL). For GCF, a higher amount of S1P was found in patients with POD (20.09 ng/mL and 15.20 ng/mL). This work raises a possible route of bone metabolism, inflammatory process, and identification of periodontitis through oral quantification of S1P, however, future studies are needed.

Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results

Catecholamine release increases in dogs with pheochromocytomas and in situations of stress. Although plasma catecholamines degrade rapidly, their metabolites, normetanephrine (NME) and metanephrine (ME), are stable in acidified urine. Our aim was to verify a human urine ELISA kit for the quantification of NME and ME in canine urine and to determine the effects on metabolite stability of sampling time (morning or midday) and day (ordinary or day spent in a clinic). We analyzed 179 urine samples from 17 healthy dogs. For NME, the mean intra-assay CV was 6.0% for all samples and 4.3% for the canine control; inter-assay CVs were 3.3, 3.8, and 12% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 90–101%. For ME, mean intra-assay CV was 6.5% for samples and 9.0% for the canine control; inter-assay CVs were 12.7, 7.2, and 22.5% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 85–89%. Dilution recovery was unsatisfactory for both metabolites. Based on our verification results, NME was selected for remaining analyses. We found no effect on NME concentrations of acidification or room temperature storage for up to 24 h. The NME:creatinine ratio was higher after the first of 3 clinic days compared to the same morning (111.2 ± 5.5 vs. 82.9 ± 5.3; p < 0.0001), but not on the other days. NME verification results were generally superior to ME. Dilution studies were unsatisfactory for both metabolites. Given that NME was stable without acidification at room temperature, urine samples can be collected at home. The clinic environment can cause higher NME:creatinine ratios, especially in unaccustomed dogs.