Fazilet Erman

A comparison of leptin and ghrelin levels in plasma and saliva of young healthy subjects

In the last 10 years, saliva has been increasingly used as a diagnostic fluid and in predictions of disease progression. Leptin and ghrelin are synthesized in several tissues including the salivary glands. The action of ghrelin is antagonistic to that of leptin. This study was undertaken to measure and compare the saliva ghrelin-leptin and plasma ghrelin-leptin levels in healthy young subjects. In 30 healthy subjects, after an overnight fast, saliva and plasma leptin levels were measured using the ELISA method while saliva and plasma immunoreactive ghrelin levels were measured using a commercial radioimmunoassay (RIA). The latter uses 125I-labeled bioactive ghrelin as a tracer and a rabbit polyclonal antibody raised against full-length octanoylated human ghrelin (Phoenix, Europe, Karlsruhe, Germany). The results of this investigation revealed that saliva leptin levels (6.19+/-2.10 microg/l) were lower than plasma levels (7.39+/-3.23 microg/l) while saliva ghrelin levels (188.5+/-84.7 pg/ml) were higher than plasma levels (126.4+/-38.5 pg/ml), when male and female subjects were considered together. Saliva leptin levels (5.93+/-1.94 microg/l) were lower than plasma levels (6.22+/-2.92 pg/ml) while saliva ghrelin levels (190.3+/-80.2 pg/ml) were higher than plasma levels (120.4+/-35.7 pg/ml) in young males. Saliva leptin levels (6.47+/-2.29 microg/l) were lower than plasma levels (8.73+/-3.14 microg/l) while saliva ghrelin levels (183.2+/-90.2 pg/ml) were higher than plasma levels (129.3+/-42.8 pg/ml) in young females, and both saliva and plasma leptin levels were slightly lower in male subjects in comparison with female subjects. Also, Immunohistochemistry study indicated that ghrelin positivity was found in ductus epithelium of salivary gland. We have demonstrated for the first time that saliva ghrelin levels were higher than in plasma while saliva leptin levels were almost the same as in plasma. Measurements of ghrelin and leptin in saliva is non-invasive, simple, and generally much preferred by patients and thus may be an acceptable alternative to plasma sampling.

Presence of obestatin in breast milk : Relationship among obestatin, ghrelin, and leptin in lactating women

OBJECTIVE The peptide hormones ghrelin and leptin have been found in blood and breast milk. This study was undertaken to investigate whether breast milk also contains obestatin, which is derived from the same gene as ghrelin but has opposite actions, and to characterize the relations among serum and milk ghrelin, obestatin, and leptin levels in lactating mothers. METHODS Venous blood, colostrum, and mature milk were obtained from healthy lactating women (n = 31) just before suckling. The ghrelin and obestatin concentrations were determined by radioimmunoassay. Leptin levels were measured by enzyme-amplified sensitivity immunoassay. RESULTS Obestatin levels in colostrum (538.9 pg/mL) and mature milk (528.5 pg/mL) were more than twice the corresponding blood levels (270.3 and 289.4 pg/mL, respectively). In contrast, leptin levels in colostrum (2.01 ng/mL) and mature milk (2.04 ng/mL) were more than five-fold lower than the corresponding blood levels (11.54 ng/mL). There was no correlation between breast milk ghrelin levels and leptin (r = -0.18, P > 0.05). However, there was a positive correlation between leptin levels in breast milk and blood (r = 0.369, P < 0.05). CONCLUSION The origin of milk obestatin is not currently known, but it comes from the blood or breast and may drain through the mammary glands into the milk. Ghrelin, obestatin, and leptin in the milk may directly affect appetite and their levels may be related to the regulation of energy balance and the pathogenesis of obesity.

Alteration in chromogranin A, obestatin and total ghrelin levels of saliva and serum in epilepsy cases☆

This study was designed to measure the levels of chromogranin A (CgA), ghrelin and obestatin in serum and saliva (including CgA expression in healthy tissue) in epileptic patients to determine any significant differences between these patients and healthy controls. Samples were obtained from a total of 91 subjects: 10 newly-diagnosed primary generalized epilepsy (PGE) patients who had started treatment with valproic acid and phenytoin for seizure control; 18 PGE patients who were previously and currently receiving treatment with valproic acid and phenytoin for seizure control; 37 patients with partial epilepsy (PE) (simple, n=17 or complex, n=20) who had been and were still being treated with carbazebime for seizures; and 26 healthy controls. CgA immunoreactivity in healthy salivary gland was analyzed by immunohistochemistry and ELISA. The levels of CgA, total ghrelin and obestatin in serum and saliva were measured by ELISA. The results revealed that normal salivary gland produces its own CgA. Before treatment, CgA levels in saliva and serum were significantly greater in patients newly-diagnosed with PGE than controls. Ghrelin and CgA concentrations were also greater in PGE patients previously or currently treated with drugs, and in patients with simple or complex partial epilepsy (PE) previously or currently treated with drugs, than in healthy normal controls. In conclusion, salivary concentrations of CgA, ghrelin and obestatin were similar to their serum levels, so saliva might be a desirable alternative to serum for measuring these hormones because it is easy and painless to collect.

Pemetrexed ameliorates experimental arthritis in rats.

Pemetrexed (PMTX) is an anti-folate drug as methotrexate. The purpose of this study was to assess the efficacy of PMTX on collagen-induced arthritis (CIA). Forty Wistar albino rats were randomized into four groups. Arthritis was induced by intradermal injection of chicken type II collagen combined with incomplete Freund’s adjuvant. Animals were sacrificed at the 15th day after the onset of arthritis. Tumor necrosis factor alpha (TNF-α), interleukin (IL)-17, and malondialdehyde (MDA) levels were increased, and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities and the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were decreased in the arthritis group. In the PMTX-treated (0.2 and 1 mg/kg/week i.p.) groups, the levels of TNF-α, IL-17, and MDA were decreased; the activities of SOD, CAT, and GPx and the expressions of Nrf2 and HO-1 were restored, and perisynovial inflammation and cartilage–bone destruction were decreased. PMTX has anti-arthritic potential in the CIA model and may be a therapeutic agent for rheumatoid arthritis.

Saliva/serum ghrelin, obestatin and homocysteine levels in patients with ischaemic heart disease

Summary Background: We aimed to compare ghrelin, obestatin, homocysteine (Hcy), vitamin B12 and folate levels in the serum and saliva of ischaemic heart disease patients. Methods: Serum and saliva were collected from 33 ischaemic heart disease (IHD) patients and 28 age- and body mass index-matched healthy individuals. Levels of acylated and desacylated ghrelin, obestatin and Hcy were determined using the ELISA method. Results: Acylated ghrelin, desacylated ghrelin and obestatin levels in the saliva were found to be higher than those in the serum of the control group, while acylated and desacylated ghrelin levels in the saliva were significantly lower than those in the serum. Obestatin levels were higher in IHD patients (p = 0.001). Saliva and serum vitamin B12 and folate levels in IHD patients were significantly lower than in the control group (p = 0.001). Conclusions: It was determined that serum ghrelin levels increased in ischaemic heart disease patients, while serum levels of obestatin decreased.

Des-Acylated Ghrelin, Rather Than Acylated Ghrelin, Might Be More Valuable in Inflammatory Bowel Diseases

To the Editor We read with interest the article entitled ‘‘Serum Ghrelin Levels in Inflammatory Bowel Disease with Relation to Disease Activity and Nutritional Status’’ by Ates et al. that was recently published in Dig Dis Sci [1]. They reported the increase in acylated ghrelin in serum from patients with active ulcerative colitis (UC) and Crohn’s disease (CD) compared with those in corresponding serum samples of patients in remission. Even though the data seem interesting and very novel, and the outcomes are clear in their study, we strongly believe that there are some important points that need to be addressed and clarified. First of all, it is unclear whether they investigated the serum or plasma ghrelin level from patients with UC and CD even though serum ghrelin levels are mentioned in their title. In our opinion, they investigated the plasma not serum ghrelin level since in their Methods section they state that blood was directly drawn into a tube bearing ethylenediaminetetraacetic acid (EDTA), and EDTA or another blood anticoagulant is used for the preparation of plasma not serum samples. As is known, serum is a clear liquid without fibrinogen and other clotting factors, so that serum is different from plasma [2]. The second point we would like to address is that in such a comparative study the measuring acylated ghrelin alone cannot go to the right address. As is known, ghrelin 28-amino acid peptid has a modification on its third residue, which is a serine. The hydroxyl group of Ser3 is post-translationally octanoylated (acyl addition). This modification seems to be essential for growth hormone release and appetite [3]. It was also known that the des-acylated form of ghrelin stimulated cell proliferation, adipogenesis [4], and counteracted the metabolic action of acylated ghrelin [5] as well as circulated in far greater amounts than the acylated form. If so, in term of inflammatory bowel diseases (ulcerative colitis and Crohn’s disease), the des-acylated form of ghrelin might be more physiologically important because inflammation in the gut is associated with increased epithelial cell proliferation [6]. Finally, to clarify a link with ghrelin and inflammatory bowel diseases, we would like to suggest that two forms of ghrelin should be tested in future experiments to obtain more reliable results.

THU0658 WNT/β-catenin pathway is affected in primary sjÖgren's syndrome

Background Sjögrens syndrome (SS) is a chronic autoimmune disease that causes salivary and lacrimal gland dysfunction, resulting in oral and ocular dryness. The pathogenesis of SS is still unknown. The Wingless (Wnt)/β-catenin pathway has been recently shown to play an important role in inflammation. Objectives The aim of the present study was to determine serum and salivary levels of Dickkopf-related protein 1 (DKK1) and sclerostin those are inhibitor of Wnt/β-catenin signaling pathway and to evaluate the expression of Wnt-1 and Wnt-3a in the salivary gland, in patients with primary SS. Methods 30 patients with primary SS, 30 patients with systemic lupus erythematosus (SLE) and 29 healthy controls were enrolled in the study. Fasting blood and saliva samples were obtained from the participants. Serum and salivary levels of DKK1 and sclerostin were measured by enzyme-linked immunosorbent assay. Wnt-1 and Wnt-3a expression were also immunohistochemically assessed in salivary gland. EULAR SS Disease Activity Index (ESSDAI), and EULAR SS Patient Reported Index (ESSPRI) in the SS group and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) in the SLE group were recorded. Results Serum DKK1 and sclerostin levels were decreased in the SS and SLE groups compared to the controls (Table 1) (p<0.001 for both). Salivary sclerostin levels were similar among the study groups (p>0.05 for all). Salivary DKK1 levels were higher in the SS group compared to the control and SLE group (p=0.004 and p=0.009, respectively). Moreover, serum DKK1 level was higher in the SS group than in the SLE group (p=0.046). Serum DKK1 level was positively correlated with serum sclerostin level in the SS, SLE and control groups (r=0.677; p<0.001, r =0.783; p<0.001, and r=0.829; p<0.001, respectively). ESSPRI was negatively correlated with serum DKK1 and sclerostin levels (r=-0.363; p=0.049 and r=-0.416; p=0.022, respectively) in the SS group. Moreover, in the salivary gland tissues, the positivities of Wnt-1 (71.4% vs. 46.2%, p=0.182) and Wnt-3a (71.4% vs. 53.8%, p=0.345) were relatively higher in the SS group compared to the control group, respectively.Table 1. Demographics and clinical variables in the study groups HC SLE SS P1 P2 P3 (SLE vs. HC) (SS vs. HC) (SS vs. SLE) Serum DKK1, ng/ml 49.8±14.9 27.8±11.8 35.2±8.8 <0.001 <0.001 0.046 Salivary DKK1, ng/ml 30.6±5.9 31.1±6.9 36.3±6.9 0.944 0.004 0.009 Serum Sclerostin, ng/ml 12.9±5.1 5.7±4.1 5.5±3.8 <0.001 <0.001 0.984 Salivary Sclerostin, ng/ml 16.1±3.4 15.7±2.5 15.6±3.7 0.877 0.838 0.996 Conclusions According to the best of our knowledge, this study is the first study evaluating the activity of Wnt/β-catenin pathway in the primary SS. The altered serum levels of DKK1 and sclerostin in primary SS and SLE is suggest that Wnt/β-catenin pathway is affected in these inflammatory diseases. Salivary DKK1 level is increased in primary SS in contrast to SLE. On the other hand, Wnt-1 and Wnt-3a expressions on the salivary gland are increased in primary SS. Therefore, it may be concluded that Wnt/β-catenin pathway acts pathogenic roles on the glandular inflammation. Disclosure of Interest None declared

Fonksiyonel dispepside obestatin ve ghrelinin rolü

Background and Aims: Functional dyspepsia is the sense of pain/discomfort in the upper abdominal region that is not due to any organic, systemic or metabolic cause. In this study, we aimed to evaluate the change in serum and salivary ghrelin and obestatin levels in functional dyspepsia. Materials and Methods: A study group of 30 functional dyspepsia patients diagnosed according to Rome III criteria and 30 healthy individuals were included. Active ghrelin, inactive ghrelin and obestatin levels were determined. Serum lipid profile was measured and analyzed statistically. Results: Functional dyspepsia patients and the control group had similar demographic characteristics, lipid profile and Helicobacter pylori infection rates. Serum active (acyl), inactive (desacyl) and total ghrelin levels of functional dyspepsia patients were significantly higher than those of the control group (p=0.004, p