BACKGROUND: Alcohol use disorder (AUD) is a relapsing disease described as excessive use of alcohol. Evidence of the role of DNA methylation in addiction is accumulating. Ghrelin is an important peptide known as appetite hormone and its role in addictive behavior has been identified. Here we aimed to determine the methylation levels of two crucial genes (GHRL and GHSR) in ghrelin signaling and further investigate the association between methylation ratios and plasma ghrelin levels.
METHODS: Individuals diagnosed with (n = 71) and without (n = 82) AUD were recruited in this study. DNA methylation levels were measured through methylation-sensitive high-resolution melting (MS-HRM). Acylated ghrelin levels were detected by ELISA. The GHRL rs696217 polymorphism was analyzed by the standard PCR-RFLP method.
RESULTS: GHRL was significantly hypermethylated (P < 0.0022) in AUD between 25 and 50% methylation than in control subjects but no significant changes of GHSR methylation were observed. Moreover, GHRL showed significant positive correlation of methylation ratio between 25 and 50% with age. A significant positive correlation between GHSR methylation and ghrelin levels in the AUD group was determined (P = 0.037). The level of GHRL methylation and the ghrelin levels showed a significant association in the control subjects (P = 0.042).
CONCLUSIONS: GHSR and GHRL methylation levels did not change significantly between control and AUD groups. However, GHRL and GHSR methylations seemed to have associations with plasma ghrelin levels in two groups. This is the first study investigating the DNA methylation of GHRL and GHSR genes in AUD.

BACKGROUND: Biliary obstruction is a relatively common condition that affects approximately 5 in 1000 people annually. Malnutrition is very common in patients with biliary obstruction and since it is associated with significant morbidity and mortality, it is important to identify factors and mechanisms involved in its development.
OBJECTIVE: To determine the influence of obstructive jaundice on the hormones controlling appetite and nutritive status.
METHODS: This was a prospective case control study performed in a tertiary center in Zagreb, Croatia. Patients with biliary obstruction undergoing internal biliary drainage from September 2012 until August 2013 were enrolled. After excluding patients who developed procedure related complications or were lost in the follow-up, out of initial 73 patients, 55 patients were included in the analysis, including 34 with benign and 21 with malignant disease. Meanwhile, 40 non-jaundiced controls were also included. Appetite, nutritional status, and serum ghrelin, cholecystokinin (CCK), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α) were determined at admission, 48 h and 28 d after internal biliary drainage. Chi square test was used for categorical variables. Continuous variables were analysed for normality by Kolmogorov-Smirnov test and relevant non-parametric (Mann-Whitney, Kruskal-Wallis, and Friedman) or parametric (t-test and analysis of variance) tests were used.
RESULTS: Patients with obstructive jaundice were significantly malnourished compared to controls, regardless of disease etiology. Plasma ghrelin and CCK levels were significantly higher in patients with obstructive jaundice. Serum bilirubin concentrations were negatively correlated with ghrelin levels and positively correlated with TNF-α, but had no correlation with CCK concentrations. After internal biliary drainage, a significant improvement of nutritional status was observed although serum concentrations of ghrelin, IL-6, and TNF-α remained significantly elevated even 28 d after the procedure. CCK levels in patients without malnutrition remained elevated 28 d after the procedure, but in patients with malnutrition, CCK levels decreased to levels comparable with those in the control group. We have not established any correlation between appetite and serum levels of ghrelin, CCK, IL-6, and TNF-α before and after biliary drainage.
CONCLUSIONS: Possible abnormalities in ghrelin and CCK regulation may be associated with the development of malnutrition during the inflammatory response in patients with biliary obstruction.

OBJECTIVE: Low resting metabolic rate (RMR), as a risk factor for weight gain and obesity, can be influenced by many factors. Empirical research has confirmed the role of appetite and related hormones in obesity and energy intake. This study aimed to investigate the relationship between appetite and related hormones in overweight or obese Iranian women with normal and hypo RMR.
METHODS: This case-control study was conducted on 42 Iranian adult women (21 cases, and 21 controls), aged 18-48 years. An impedance body analyzer was used to obtain the body composition and an indirect calorimeter was used to assess the RMR. The Flint questionnaire was used to assess appetite, dietary intake, and physical activity were assessed by FFQ and IPAQ questionnaires respectively, and ELISA kits were used to assess leptin, ghrelin, and insulin hormones.
RESULTS: The results of the study demonstrated a negative association between ghrelin hormone level (β = -0.34, 95%CI = -61.70,-3.86, P-value = 0.027) and RMR, and a positive association between insulin hormone level (β = 0.48, 95%CI = 9.38-34.35, P-value = 0.001) and RMR. Also, results of the appetite questionnaire showed that, in general, both appetite (β = 0.32, 95%CI = -0.10-2.99 P-value = 0.044) and hunger variable (β = 0.30, 95%CI = 0.04-5.87, P-value = 0.047) have a positive association with RMR. There was no significant association between leptin levels and RMR.
CONCLUSIONS: It is evident that appetite and related hormones have a potential role in promoting a normal RMR.

OBJECTIVE: The aim of the study was to discover the interrelation between the severity of gastroesophageal reflux disease (GERD) symptoms, acid exposure time (AET), excessive daytime sleepiness (EDS) and the level of active blood plasma ghrelin in the patients with undifferentiated connective tissue disease (UCTD).
METHODS: Materials and methods: The study included 120 patients with GERD. All the patients were divided in two groups: Group I - GERD was not accompanied by the signs of connective tissue disease (n=45) and Group II - GERD developed on the background of UCTD syndrome (n=75). Daily transnasal pH monitoring was performed to determine the nature of pathological refluxes. EDS was detected by The Epworth Sleepiness Scale. Active ghrelin in blood plasma samples was determined by ELISA.
RESULTS: Results: 80% of the patients of Group II and 35.48% of Group I suffered from EDS (p<0.05). The mean daily AET index was 5.48±0.4% in Group II and 6±0.2% in Group I, in the night hours mostly when patients were in the upright position. This phenomenon contributed to a deterioration of sleep quality and the appearance of EDS and was supported by a connection between AET and EDS (r=+0.827 for Group I and r=+0.768 for Group II). The mean De Meester index was higher in the patients of Group II (23.01±2.24 in Group I vs 31.08±2.4 in Group II; p<0.05).
CONCLUSIONS: Conclusions: GERD manifestations are strongly related to the level to AET and intensity of EDS. The EDS symptoms depend on circulating ghrelin level.

The objective of this study was to evaluate the relationship between food intake and serum levels of leptin and ghrelin in the luteal (LP) and follicular (FP) phases of the MC (menstrual cycle) in participants with and without PMS (premenstrual syndrome).
This was a case-control study with healthy participants aged 20-45 years with regular menstrual cycles (24-35 days) with and without PMS. After the Daily Record of Severity of Problems (DRSP) was filled out for two months (PMS diagnosis), a nutritional assessment was carried out based on twelve food intake records (for two menstrual cycles) to quantify food intake.
Of the 69 participants analyzed, 35 experienced PMS and 34 did not experience PMS. For participants with PMS, calorie and carbohydrate intake was higher during LP than in FP (p = 0.004 and p = 0.003, respectively), whereas these changes were not observed in participants without PMS (p > 0.05). There were interactions between the groups and the MC phases (LP and FP) for the intake of calories (p = 0.028) and carbohydrates (p = 0.001). There was a marginal negative relationship between the levels of ghrelin and calorie intake in FP (rS = -0.314, p = 0.066) in the PMS group and a negative relationship between the levels of ghrelin and leptin in LP (rS = -0.490, p = 0.004) in the group without PMS.
These results indicated a higher calorie and carbohydrate intake during LP in participants with PMS, in addition to the hypothesis that the roles of ghrelin and leptin in energy regulation may be different in participants with PMS compared to those without PMS.

The nutrient composition of breast milk alters during lactation, and maternal BMI adds more intricacy into its complexity. We aimed to compare leptin, ghrelin, adiponectin and insulin-like growth factor-1 (IGF-1) levels of pre-feed and post-feed breast milk in mothers with obesity and normal weight, and tried to determine their effects on infants\' growth over weight for length z-score. Twenty obese and twenty normal weight mothers with 2-month-old infants were enrolled in this case-control study. Five millilitre pre-feed breast milk and 5 ml post-feed breast milk were collected. Breast milk leptin, ghrelin, adiponectin and IGF-1 were measured by commercial kits. The pre-feed breast milk of mothers with obesity had significantly higher levels of ghrelin than mothers with normal weight (P = 0·025), whereas the post-feed breast milk of mothers with normal weight had higher levels of adiponectin than the mothers with obesity (P = 0·010). No significant differences were observed in leptin and IGF-1 levels between the two groups. Post-feed breast milk IGF-1 levels of mothers with obesity were correlated with infant\'s weight for length z-score at 2 months (r -0·476; P = 0·034). In linear regression models, parity affected the ghrelin in pre-feed breast milk (P = 0·025). Our results revealed that maternal pre-pregnancy BMI was associated with breast milk components.

The global incidence of breast cancer among men is steadily growing. Despite this, compared to female breast cancer patients, there are very few studies on biomarkers in male breast cancer patients. A cross-sectional case control study was carried out to determine the serum levels of melatonin, ghrelin, dopamine, serotonin, epinephrine, and GABA in male breast cancer. All the recruited patients were obese, old, and had recently been diagnosed with the disease. They had not received any treatment for the cancer until the time of the study. Melatonin and epinephrine serum levels were significantly higher in breast cancer patients compared to their age-matched controls, whereas ghrelin, dopamine, GABA, and serotonin serum levels were lower in patients compared to the control group. The serum levels of most of the studied biomarkers in male breast cancer patients were similar to those observed in female breast cancer patients, except for serum melatonin levels.

Natural ghrelin, a peptide growth hormone secretagogue, has a therapeutic potential in cachexia. We designed a dose-finding trial of subcutaneous natural ghrelin to improve nutritional intake (NI) in advanced cancer patients.
Advanced cancer patients with cachexia management (symptom management, physiotherapy, nutritional, and psychosocial support) started with ghrelin at 32 μg/kg body weight, followed by 50% dose increases. Patients self-injected ghrelin twice daily for 4 days followed by a wash-out period. After reaching the primary endpoint, maximal NI (minimal dose for maximal NI), a maintenance period followed during which patients injected 10 doses of ghrelin per week. Safety parameters, NI, and cachexia outcomes (symptoms, narratives, muscle mass, and strength) were measured over 6 weeks.
Ten patients with metastatic solid tumours were included, and six (100% male, mean age 61.8 ± 8.5 SD) received ghrelin. Minimal dose for maximal NI was reached in four patients. Three patients reached the end-of study visit. Ghrelin was well tolerated with variable results on appetite and eating-related symptoms but a positive effect in the narratives. Mean Functional Assessment of Appetite & Cachexia Therapy score was 6.8 points lower at final measurement compared with baseline, t(5) = 5.98, P < .01. Muscle mass was stable in two patients and increased in one patient, and muscle strength was stable in three patients. Subjective tolerability was high. Patients showed a fluctuating trajectory, and median survival was 88 days (51-412 days).
Ghrelin was safe in advanced patients with cancer cachexia without dose-limiting toxicity and well tolerated. The intervention was very complex, and the number of patients included was small. There was a positive effect on nutritional intake and patient narratives.

BACKGROUND: Adipokines are emerging mediators of immune response, and may affect susceptibility to active TB.OBJECTIVE: To examine the associations between adipokines and the risk of active TB.METHODS: In a case-control study nested within a prospective cohort of middle-aged and older adults in Singapore, 280 incident active TB cases who donated blood for research before diagnosis were matched with 280 controls. Serum levels of adiponectin, resistin, leptin and ghrelin were measured. Multivariable logistic regression models were used to compute the adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the associations between adipokines and the risk of active TB.RESULTS: Higher levels of leptin and resistin were associated with reduced risk of TB in a dose-dependent manner. Compared to those in the lowest quartile of leptin levels, those in the highest quartile had an OR of 0.46 (95%CI 0.26-0.82; P for trend = 0.009). Similarly, compared to those in the lowest quartile of resistin levels, those in the highest quartile had an OR of 0.46 (95%CI 0.24-0.90; P for trend = 0.03). Adiponectin and ghrelin levels were not associated with TB risk.CONCLUSION: Increased serum levels of leptin and resistin may be associated with reduced susceptibility to active TB infection.

Background Ghrelin and obestatin are two gastric hormones encoded by the same preproghrelin gene that convey information concerning nutritional status to the central nervous system. Ghrelin has been considered as an appetite stimulating peptide that has a role in the regulation of energy homeostasis. Obestatin has been described for its appetite suppressing effects opposing ghrelin\'s effect on food intake. The study aimed to evaluate ghrelin, obestatin and the ghrelin/obestatin ratio in obese children compared to non-obese and correlate them to food macronutrients intake. Methods This study is a cross-sectional case control study comprising 60 obese children, in addition to 31 age- and sex-matched controls. All children were subjected to clinical examination, anthropometric assessment, and a 3-day 24-h dietary recall. Fasting serum ghrelin and obestatin levels were evaluated, the ghrelin/obestatin ratio was calculated and they were correlated to macronutrients intake. Results Obese children had significantly lower serum fasting levels of ghrelin, obestatin and the ghrelin/obestatin ratio than the control group. The mean intake of total energy and macronutrients was significantly higher in obese children. Ghrelin showed positive correlation with total energy and fat intake in the obese group. Obestatin had positive correlations with total energy and fat intake while the ghrelin/obestatin ratio had a negative correlation with the total energy intake in the control group. Conclusions Ghrelin, obestatin and the ghrelin/obestatin ratio were significantly lower in obese children and significantly associated with their total energy intake. Disturbed ghrelin to obestatin balance may have a role in the etiology and pathophysiology of obesity.