The role of trimethylamine-N-oxide level in the diagnosis of diabetic retinopathy and the differential diagnosis of diabetic and nondiabetic retinopathy / Papel do nível de N-óxido de trimetilamina no diagnóstico da retinopatia diabética e no diagnóstico diferencial da retinopatia diabética e não-diabética

ABSTRACT Purpose: Trimethylamine N-oxide serum levels have been associated with type 2 diabetes mellitus and its complications. The current study aimed to find out if plasma trimethylamine N-oxide level may be a novel marker in the diagnosis of diabetic retinopathy and if it can be used in the differential diagnosis of diabetic and nondiabetic retinopathy. Methods: The study included 30 patients with diabetic retinopathy, 30 patients with nondiabetic retinopathy, 30 patients with type 2 diabetes mellitus without retinopathy, and 30 healthy control participants. Biochemical parameters, serum IL-6, TNF-α, and trimethylamine N-oxide levels were measured in all participants. Results: Trimethylamine N-oxide level was significantly higher in diabetic retinopathy than in the other groups (p<0.001). There was no significant difference in trimethylamine N-oxide levels between nondiabetic retinopathy and control or type 2 diabetes mellitus Groups. There was a significant positive correlation between trimethylamine N-oxide level and elevated FPG, BMI, HOMA-IR score, BUN, IL-6, and TNF-α levels. Conclusion: The current study showed that the trimethylamine N-oxide level is elevated in diabetic retinopathy. These findings suggest that serum trimethylamine N-oxide level might be a novel marker for diabetic retinopathy, and it might be used in the differential diagnosis of diabetic and nondiabetic retinopathy.

RESUMO Objetivo: Os níveis séricos de N-óxido de trimetilamina têm sido associados ao diabetes mellitus tipo 2 e suas complicações. O presente estudo tem como objetivo responder a duas questões, entre elas: O nível plasmático de N-óxido de trimetilamina poderia ser um novo marcador no diagnóstico de retinopatia diabética? e Ele poderia ser utilizado no diagnóstico diferencial de retinopatia diabética e não diabética? Métodos: Trinta pacientes com retinopatia diabética, 30 pacientes com retinopatia não diabética, 30 pacientes com diabetes mellitus tipo 2 sem retinopatia e 30 participantes saudáveis do grupo controle foram incluídos no estudo. Parâmetros bioquímicos, níveis séricos de IL-6, de TNF-α e de N-óxido de trimetilamina foram medidos em todos os participantes. Resultados: O nível de N-óxido de trimetilamina foi significativamente maior na retinopatia diabética do que nos outros grupos (p<0,001). Não houve diferença significativa no nível de N-óxido de trimetilamina entre o grupo de retinopatia não diabética, do grupo controle ou do grupo de diabetes mellitus tipo 2. Houve uma correlação positiva significativa entre o nível de N-óxido de trimetilamina e os níveis elevados de FPG, IMC, HOMA-IR, BUN, IL-6 e TNF-α. Conclusão: O estudo atual mostrou que o nível de N-óxido de trimetilamina encontra-se elevado na retinopatia diabética. Esses achados sugerem que o nível sérico de N-óxido de trimetilamina pode ser um novo marcador na retinopatia diabética, podendo ser usado no diagnóstico diferencial de retinopatia diabética e não diabética.

Evaluation of Serum ANGPTL8/Betatrophin and Cartonectin/CTRP3 Levels in Diabetic and Non-Diabetic Retinopathy.

OBJECTIVE: To determine serum betatrophin and cartonectin levels and their relationship with biochemical parameters in diabetic and non-diabetic retinopathy. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Internal Medicine and Ophthalmology, School of Medicine, Firat University, Turkey, from April to November 2020. METHODOLOGY: Patients with diabetic retinopathy (DR), non-diabetic retinopathy (non-DR), type 2 diabetes mellitus without retinopathy, and healthy controls, were enrolled from April to November 2020. Demographic, metabolic, and biochemical parameters were evaluated. Serum betatrophin, cartonectin, IL-6, and TNFα levels were assayed by ELISA methods. One-Way Anova or Kruskal Wallis tests were applied to find out statistical significance among different variables between four groups. RESULTS: A total of 84 participants (DR= 21 patients; non-DR= 21 patients; 21 diabetic patients without retinopathy and 21 healthy controls) were enrolled in the study. TNF-α level was significantly higher in both DR and non-DR than in controls (p<0.001). IL-6 level was higher in DR group than T2DM and controls (p=0.013). Serum betatrophin level was significantly higher in DR group than in non-DR and T2DM groups (p=0.002). Cartonectin level was decreased in DR, non-DR, and T2DM groups compared to non-diabetic healthy controls (p=0.002). CONCLUSION: Serum betatrophin levels are higher in DR, whereas cartonectin levels are lower in both DR and non-DR groups. Serum betatrophins and cartonectin estimation may be helpful in early diagnosis and differential diagnosis in cases of diabetic and non-diabetic retinopathy. KEY WORDS: Angiopoietin-like protein 8, Cartonectin/CTRP3, Diabetic retinopathy, Diabetes mellitus, Type 2, Diagnosis, Differential, Betatrophin.

The role of trimethylamine-N-oxide level in the diagnosis of diabetic retinopathy and the differential diagnosis of diabetic and nondiabetic retinopathy.

PURPOSE: Trimethylamine N-oxide serum levels have been associated with type 2 diabetes mellitus and its complications. The current study aimed to find out if plasma trimethylamine N-oxide level may be a novel marker in the diagnosis of diabetic retinopathy and if it can be used in the differential diagnosis of diabetic and nondiabetic retinopathy. METHODS: The study included 30 patients with diabetic retinopathy, 30 patients with nondiabetic retinopathy, 30 patients with type 2 diabetes mellitus without retinopathy, and 30 healthy control participants. Biochemical parameters, serum IL-6, TNF-α, and trimethylamine N-oxide levels were measured in all participants. RESULTS: Trimethylamine N-oxide level was significantly higher in diabetic retinopathy than in the other groups (p<0.001). There was no significant difference in trimethylamine N-oxide levels between nondiabetic retinopathy and control or type 2 diabetes mellitus Groups. There was a significant positive correlation between trimethylamine N-oxide level and elevated FPG, BMI, HOMA-IR score, BUN, IL-6, and TNF-α levels. CONCLUSION: The current study showed that the trimethylamine N-oxide level is elevated in diabetic retinopathy. These findings suggest that serum trimethylamine N-oxide level might be a novel marker for diabetic retinopathy, and it might be used in the differential diagnosis of diabetic and nondiabetic retinopathy.

m6A RNA, FTO, ALKBH5 Expression in Type 2 Diabetic and Obesity Patients.

OBJECTIVE: To investigate whether m6A content changes in type 2 diabetes mellitus (T2DM) and obese individuals and whether the relationship of m6A content with the mRNA expression levels of FTO and ALKBH5 genes. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Department of Internal Medicine, Firat University, Medical School, Elazig, Turkey, between January 2019 and January 2022. METHODOLOGY: The study included 34 newly diagnosed patients with type 2 diabetes mellitus, 34 obese individuals, and 33 healthy individuals without any chronic and metabolic disease matched for age and gender. The global m6A RNA methylation, FTO, and ALKBH5 gene analyses of all the participants were performed. Total cholesterol, triglyceride, LDL, HDL, HbA1c, and insulin and glucose levels were measured. RESULTS: The median percentages of m6A RNA methylation in the control group, obese, and T2DM participants were 5.62%, 4.20%, and 5.21% respectively (p=0.004). The m6A RNA methylation percentage of the obese participants was significantly lower than controls (p=0.021). The FTO and ALKBH5 mRNA levels were significantly lower in obese and T2DM participants than in controls. There was a negative significant correlation between m6A RNA level and FTO i.e. (r=-0.291, p=0.003) and ALKBH5 (r=-0.321. p=0.001) levels. CONCLUSION: m6A RNA expression levels of obese individuals were lower than healthy controls. The FTO and ALKBH5 mRNA expressions were lower in both obese and T2DM participants compared to the healthy controls. There was no significant difference between obese and T2DM individuals in terms of m6A RNA expression, FTO and ALKBH5 mRNA expression. m6A RNA expression, FTO, and ALKBH5 levels have a potential role in obesity and diabetes mellitus. KEY WORDS: m6A RNA methylation, Epigenesis, Genetic, FTO, ALKBH5.

Role of Betatrophin and Inflammation Markers in Type 2 Diabetes Mellitus, Prediabetes and Metabolic Syndrome.

OBJECTIVE: To determine the association of betatrophin and inflammation factors in metabolic diseases such as diabetes, impaired fasting glucose, impaired glucose tolerance and metabolic syndrome. STUDY DESIGN: A cross-sectional, analytical study. PLACE AND DURATION OF STUDY: Firat University Medical School between April 2017 and December 2020. METHODOLOGY: The study included 20 patients with type 2 diabetes mellitus, 20 patients with impaired fasting glucose (IFG), 20 patients with impaired glucose tolerance (IGT), 20 patients with metabolic syndrome (MetS), and a control group consisting of 20 healthy individuals. Anthropometric, fasting serum biochemical data were collected. Circulating betatrophin, and inflammation markers were measured by enzyme-linked immunosorbent assay (ELISA). The association of betatrophin, TNF-alpha, and IL-6 levels between groups were performed with One-way ANOVA and post-hoc Tukey HSD test. RESULTS: Significantly higher levels of circulating betatrophin were observed in IFG, IGT, And MetS groups compared to healthy controls (p=0.017). There were significantly difference TNF-α levels in IFG, IGT, and MetS groups compared to healthy controls (p<0.001). The levels of IL-6 were significantly higher in MetS group than healthy controls (p=0.007). CONCLUSION: The circulating betatrophin and TNF-α levels were increased in MetS, IFG and IGT. IL-6 was decreased in MetS compared to the healty controls. Further studies are needed to elucidate the role of betatrophin and inflammatory parameters in the development of T2DM and prediabetic syndromes, whether betatrophin could have clinical applications in the development of new antidiabetic agents. Key Words: Betatrophin, IL-6, TNF-alpha, Metabolic syndrome, Insulin resistance, Diabetes mellitus type 2.

Relationship between Serum Elabela Level and Hypertensive Nephropathy.

OBJECTIVE: To determine the difference in serum Elabela level in hypertensive patients with and without nephropathy compared to the healthy control group. Study Desing:  Cross-sectional descriptive study. PLACE AND DURATION OF STUDY: Firat University Medical School, Elazig, Turkey between December 2018 and November 2020. METHODOLOGY: The cross-sectional descriptive study consisted of 37 patients with hypertensive nephropathy (group 3), 50 hypertensive patients without nephropathy (group 2), and 50 healthy controls (group 1). Hypertensive nephropathy was defined as serum creatinine ≥1.8 mg / dl or  GFR <40 ml / minute. Biochemical parameters (Glucose, AST, ALT, urea, creatinine, lipid levels, hemogram, calcium, phosphorus, parathormone) and the levels of serum Elabela were evaluated and compared. RESULTS: There was no significant difference in age (0.270) and gender (0.951) between groups. The median Elabela levels of the three groups were 40.3 ng/mL (22.5-54.6), 5.1 ng/mL (3.7-8.3), 9.2 ng/mL (6.1-23.1), respectively with a significant difference (p<0.001). CONCLUSION: The plasma levels of Elabela were lower in the case of hypertension, independent of nephropathy. However, this decrease is not specific for nephropathy and may be due to other accompanying chronic diseases. Key Words: Hypertension, Hypertensive nephropathy, Elabela.

Elabela levels in patients with type 2 diabetes: can it be a marker for diabetic nephropathy?

BACKGROUND: Elabela (ELA) is a hormone that is secreted at high levels in the kidneys of a healthy adult. This study aims to investigate whether serum ELA levels of patients with Type 2 Diabetes vary with the severity of renal damage. METHODS: Our study included 50 healthy control subjects and 100 diabetic patients, who were categorized into groups based on urine albumin/creatinine ratios (ACR). Patients included in the study were assigned to four groups: Group 1 (healthy control), Group 2 (ACR<29mg/g), Group 3 (ACR=30-299 mg/g), and Group 4 (ACR>300 mg/g normal or high serum creatinine). Physical examination findings, demographic characteristics of the study group were recorded, and serum ELA levels and other laboratory parameters were assessed using appropriate methods. RESULTS: The results of the study indicated that ELA levels determined in healthy individuals gradually decreased through stages of normal albuminuria, microalbuminuria, and macroalbuminuria. Moreover, ELA had a significant negative correlation with LDL-C (r=-0.201, p=0.014), glucose (r=-0.437, P<0.001), retinopathy (r=-0.222, P=0.006), serum BUN (r=-0.161, P=0.049), and a positive correlation with eGFR (r=0.250, P=0.002). CONCLUSIONS: The fact that ELA levels are higher in healthy individuals compared to diabetic patients without microalbuminuria, and higher in diabetic patients without microalbuminuria compared to patients with advanced albuminuria and kidney damage, suggests that the ELA level can be an important clinical prognostic variable and even a promising agent for the treatment of diabetic nephropathy patients.

Low serum levels of meteorin-like/subfatin: an indicator of diabetes mellitus and insulin resistance?

INTRODUCTION: Meteorin-like (Metrnl), also known as subfatin, is a recently discovered adipokine with a favourable effect on insulin sensitivity. Studies have shown lower Metrnl levels in obese patients. However, data on its circulating levels in type 2 diabetes mellitus (T2DM) patients are contradictory. This study aims to evaluate serum Metrnl levels in T2DM patients and determine the relationship between serum Metrnl levels and insulin resistance in these patients. MATERIAL AND METHODS: This cross-sectional study was conducted among 150 participants. The study was carried out between June 2019 and December 2019 at the internal medicine outpatient clinic of a tertiary university hospital. The participants were divided into three groups: group 1 (control group, n = 50), group 2 (newly diagnosed T2DM, n = 50), and group 3 (long-standing diagnosed T2DM, n = 50). An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of Subfatin (Metrnl), and the correlations of Metrnl level with anthropometric parameters, HOMA index, and biochemical measurements were assessed. RESULTS: There was no statistically significant difference between the gender (p = 0.468) and age (p = 0.067) characteristics of the three groups. The Metrnl (subfatin) levels of the participants were as follows: control group - 20.05 (1.56-103.78); newly diagnosed T2DM group - 2.62 (1.25-103.78); and long-standing diagnosed T2DM group - 2.01 (0.80-19.84) pg/mL. The Metrnl (subfatin) levels of the participants in the control group were higher than in the participants in the newly diagnosed T2DM and long-standing diagnosed T2DM groups (p < 0.001). Subfatin demonstrated a negative correlation with insulin and HOMA-IR in the control group and long-standing diagnosed T2DM group. CONCLUSIONS: The subfatin level was found to be higher in the healthy control group than in both diabetic patient groups. Subfatin level showed negative correlation with both insulin level and HOMA index. There was a relationship between subfatin and insulin resistance. Low levels of subfatin in the diabetic patient groups may play a role in the pathogenesis of T2DM by increasing insulin resistance.

Neutrophil and platelet to lymphocyte ratio in patients with hypothyroid Hashimoto's thyroiditis.

AIM: Chronic lymphocytic thyroiditis is among the most common causes of hypothyroidism along with HT (Hashimoto's thyroiditis) goitre, which is also named as autoimmune thyroiditis. Our study aims to determine the usefulness of PLR (platelet to lymphocyte ratio) and NLR (neutrophil to lymphocyte ratio), which can be obtained with a hemogram, at the clinical course or the severity of the disease in patients with Hashimoto's thyroiditis. MATERIALS AND METHODS: Our study is a retrospective cross-sectional study that included 121 hypothyroid or subclinical hypothyroid Hashimoto's thyroiditis patients and a healthy control group comprised of 100 individuals. Thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4), anti-thyroid peroxidase (anti-TPO), complete blood count (CBC), and C-reactive protein (CRP) results were obtained from patient files for both HT patients and the control group, and we computed PLR and NLR for both groups. RESULTS: PLR was lower in patients diagnosed with HT compared to the healthy control group, with statistical significance (respectively, 130.8±50.5 versus 145.3±58.5; p<0,05). NLR was higher in patients diagnosed with HT compared to the control group and a statistically significant relationship was determined (respectively, 2.43±0.94 versus 2,11±0,81; p<0,05). In addition to the present findings, we determined that PLR and NLR were correlated with anti-TPO, TSH, and FT4, although without statistical significance. CONCLUSION: As values that can be measured with an inexpensive and easily accessible routine hemogram, PLR and NLR can serve as practical and valuable markers at the clinical course or the severity of the disease and other diseases that are autoimmune and progress with chronic inflammation.

Effects of vitamin D on apoptosis and betatrophin in the kidney tissue of experimental diabetic rats.

The aim of this study is to investigate the effects of vitD on betatrophin and apoptosis in rat kidney tissue using an experimental diabetes model created with STZ. 41 male Wistar-albino breed rat were assigned to 5 groups, which included 3 groups consisting of 7 animals each and 2 groups consisting of 10 animals each. The control group received no treatments. Single-dose 0.1 M sodium buffer was administered ip to the Buffer group. The VitD group was orally administered 200 IU/day vitD.The Diabetes group was injected ip with single-dose 50 mg/kg STZ by dissolving the material in 0.1 M sodium buffer. Subjects with a glucose level exceeding 250 mg/dl were accepted to be diabetic. The Diabetes + VitD group was injected ip with 50 mg/kg single-dose STZ by dissolving the material in 0.1 M sodium buffer. Once diabetes was established, 200 IU/day vitamin D was administered orally. The histological and biochemical analyses of the Control, Buffer, and Vitamin D groups revealed similar serum TOS and TAS levels, and TUNEL positivity and betatrophin immunoreactivity. While the Diabetes group showed significantly higher TOS levels and TUNEL positivity compared to the Control group, their TAS levels and betatrophin immunoreactivity were significantly reduced. The Diabetes+Vitamin group demonstrated significantly lower TOS levels and TUNEL positivity compared to the Diabetic group, and their TAS levels and betatrophin immunoreactivity increased significantly.In conclusion; experimental diabetes was found to increase TOS and apoptotic cells and decrease TAS and betatrophin levels in kidney tissue in experimental diabetes, and that administering VitD as treatment caused a decrease in TOS and apoptotic cells and an increase in TAS and betatrophin levels. It was concluded that future studies needed to investigate various experimental diabetes times so that the role of diabetes in the pathophysiology of its effect on kidney tissue could be uncovered.

Can hematological parameters in type 2 diabetes predict microvascular complication development?

OBJECTIVE: To examine potential associations between neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, mean platelet volume (MPV), HbA1c and microvascular complications in diabetic patients from a cost-effectiveness perspective. METHODS: One hundred patients with type 2 diabetes attending our outpatient unit between May 2018 and October 2018 were included, and 100 healthy individuals served as the control group. A retrospective file search was performed to collect information on hemoglobin, mean platelet volume (MPV), glycosylated haemoglobin (HbA1c), hematocrit (Hct), neutrophil and lymphocyte count, neutrophil/lymphocyte ratio (NLR), platelets (Plt), platelet/lymphocyte ratio (PLR), and microvascular complications (neuropathy, retinopathy, nephropathy). RESULTS: Demographic and laboratory data were retrospectively controlled between diabetes (n=100) and healthy control (n=100) groups. The mean age in diabetic patients and healthy controls was 56.34 and 36.68 years, respectively. The mean NLR in diabetics and healthy controls was 2.48 and 2.11, the difference in NLR being significant (p=0.002). MPV in diabetics and controls was 8.54 and 8.53, respectively, and the difference was not significant (p=0.93). PLR was also similar, i.e. 149.7 and 145.3 in diabetics and healthy controls (p=0.067). With respect to microvascular complications, retinopathy was found to be significantly associated with MPV and NLR (p=0.015, and p=0.051), and nephropathy showed a significant association with NLR (p=0.027) among diabetics. In contrast with the two other microvascular complications, no significant association between neuropathy and NLR could be detected, while PLR and neuropathy was significantly associated (p=0.003). CONCLUSION: Microvascular complications may be associated with certain hematologic parameters, as suggested by comparisons both between diabetics and healthy individuals and within the group of diabetic individuals. We believe that hematologic parameters such as hematocrit, MPV, NLR, and PLR, which can be obtained through a simple complete blood count, may be utilized as cost-effective predictors of diabetic microvascular complications. Further prospective studies with larger sample size are required to better delineate these associations.

Low prevalence of obesity in Behçet's disease is associated with high obestatin level.

OBJECTIVE: Chronic inflammatory diseases are associated with altered body composition. Ghrelin has anti-inflammatory effects, and its level is altered in obesity and inflammatory diseases. The aim of the study was to evaluate the prevalence of obesity and ghrelin and obestatin levels in patients with Behçet's disease (BD). MATERIAL AND METHODS: One hundred and forty-three (143) patients with BD and 112 healthy controls (HC) were enrolled. Participants were subdivided according to the body mass index (BMI) as lean (<18.5 kg/m2), normal weight (18.5-24.9 kg/m2), overweight (25-29.9 kg/m2) and obese (≥30 kg/m2). In addition to the routine evaluations (fasting blood glucose, lipid profile, and kidney and liver function tests), serum acylated-ghrelin (AG), unacylated-ghrelin (UAG), total ghrelin (TG) and obestatin levels were analyzed. Student's t-test and chi-square test were used for statistical analysis. RESULTS: The prevalence of obesity was relatively lower in the BD group than in the HC group (12.6% vs. 20.5%, p=0.089). Serum ghrelin levels were similar in the BD and HC groups (p>0.05 for all) although the obestatin level was higher in the BD group compared to the HC group (p<0.001). Serum UAG, TG and obestatin levels were lower in obese BD patients (n=18) than non-obese BD patients (p=0.027, p=0.014 and p=0.001, respectively). CONCLUSION: The obestatin level was high and the prevalence of obesity was low in the BD group. Moreover, obese BD patients had low obestatin levels. These results suggest that obestatin may protect BD patients from obesity.

Carotid artery stiffness in Behçet's disease.

OBJECTIVE: Increased carotid arterial stiffness (CAS) is a predictor of subclinical early atherosclerosis as well as carotid intima-media thickness (cIMT). We aimed to determine CAS and cIMT in Behçet's disease (BD). MATERIAL AND METHODS: BD (n=49) and rheumatoid arthritis (RA) (n=64) patients and healthy controls (HC) (n=40) were included in the study. cIMT was measured. CAS indices, including arterial compliance (AC), arterial distensibility (AD), Young's elastic modulus (YEM), Peterson's elastic modulus (Ep), and ß stiffness index (ßSI) were measured based on the diameter-pressure relationship. RESULTS: When compared to the HC group, the mean cIMT was significantly higher in the RA group (p=0.033), but it was not higher in the BD group. The CAS indices, including AD, AC, Ep, and ßSI were not significantly different among the study groups. Moreover, the cIMT and CAS indices were not significantly different between active (n=20) and inactive BD patients, and these indices were not correlated with the scores of disease activity. AD, AC and Ep were significantly lower in the BD patients with a positive pathergy reaction than in those with a negative reaction. CONCLUSION: These results suggest that BD does not directly lead to arterial stiffness or to an increase in cIMT.

Plasma urotensin II levels in primary Raynaud's phenomenon and systemic sclerosis

Background/aim: The pathogenesis of Raynaud's phenomenon (RP) has not yet been fully elucidated. RP is characterized by exaggerated cold-induced vasoconstriction. Urotensin II (UII) is a potent vasoconstrictor. The aim of the present study was to evaluate plasma UII levels in both primary RP and secondary RP associated with systemic sclerosis (SSc).Materials and methods: Fifteen patients with primary RP, 30 patients with RP secondary to SSc, and 30 healthy controls (HC) were included in the study. Raynaud condition scores (RCS) were determined in the primary RP and SSc groups. Modified Rodnan skin score (MRSS) was determined for the SSc patients. Plasma UII level was analyzed by the ELISA method. Results: When compared to the HC group, plasma UII level was lower in the secondary RP group, but not in the primary RP group. Plasma UII level was not directly related to RCS in either the primary or secondary RP group. Moreover, it was not correlated with MRSS in the secondary RP group.Conclusion: The results of the present study suggest that UII is not associated with primary RP. Its level was lower in the secondary RP (SSc) patients. Therefore, it can be concluded that decreased UII level is related to SSc instead of RP.

Fat-free milk as a therapeutic approach for constipation and the effect on serum motilin and ghrelin levels.

OBJECTIVE: This study explores the effects of fat-free milk supplementation on individuals with chronic constipation with regard to levels of motilin and acylated and des-acylated ghrelin (which affect intestinal motility) and compares them with data from control subjects given whole milk supplementation. METHODS: The investigation was designed according to the constipation severity test of individuals whose ages and body mass indexes were comparable. Individuals with mild constipation (n=10) were supplemented with 400 mL of fat-free milk daily; moderate constipation cases (n=10) were supplemented with 600 mL, and severe constipation cases (n=10) were supplemented with 800 mL of fat-free milk daily. Healthy control subjects were administered 400 mL of fat-free milk (group 1), which was followed a month later by administration of 400 mL of whole milk for 3 days (group 2). Blood samples were collected from the subjects before and after milk supplementation for hormone analyses. Motilin and acylated and des-acylated ghrelin were quantified with ELISA assay. RESULTS: Supplementation of fat-free milk significantly increased levels of circulating motilin and ghrelin in all groups, including the control subjects, but whole milk supplementation led to a decrease in these hormone levels in the control subjects. CONCLUSION: Drinking fat-free milk might be a new way of solving constipation.

Ghrelin and obestatin expression in oral squamous cell carcinoma: an immunohistochemical and biochemical study.

The underlying molecular mechanism of carcinogenesis in oral squamous cell carcinoma (OSCC) is poorly understood and appears to be controlled on many genetic, environmental, and hormonal factors. Obestatin and ghrelin, two recently discovered hormones, are co-expressed in endocrine cells. The purpose of this investigation was to examine the immunohistochemical features of OSCCs in relation to the tissue concentration of ghrelin and obestatin. The association between OSCC and Epstein Barr Virus (EBV) status was also explored. The expression of ghrelin and obestatin was examined by immunohistochemistry and immunoassay in oral biopsy specimens: 10 benign squamous epithelial cell samples, 10 microinvasive squamous cell carcinomas, and seven well-differentiated and seven poorly differentiated OSCCs. The presence of EBV was evaluated in these samples using immunohistochemistry. The concentrations of ghrelin and obestatin in tissue homogenates were measured by RIA and ELISA, respectively. Squamous cell carcinomas and benign tissue samples were positive for anti-EBV antibody, and obestatin and ghrelin were shown to be co-expressed in all stratified squamous epithelium samples. Expression of ghrelin and obestatin was decreased or absent in OSCCs in relation to the invasiveness of the carcinoma; ghrelin and obestatin levels in cancerous tissue homogenates were lower than in benign tissue homogenates. These results indicate that the concentrations and distribution of immunoreactive obestatin and ghrelin might be helpful in distinguishing OSCC from benign tumors. Maintaining normal levels of these hormones might be required for regulation of normal cell division. However, detailed studies will be required for better understanding of the complex mechanism of carcinogenesis relating to OSCCs.

Reversal of ergotamine-induced vasospasm following methylprednisolone.

INTRODUCTION: Ergotamine, an ergot alkaloid with partial agonist effects on alpha1 receptors and serotonin receptors, is widely used in the treatment of migraine. Ergotamine may cause severe vasospasm. CASE REPORT: A 25-year-old man was admitted to the emergency department with complaints of sudden coldness, pallor, and pain in his hands and feet for 2 days. He had been using a drug containing ergotamine for his migraine headaches for 1 week. On examination, the pulses of the radial, ulnar, popliteal, and tibial arteries were bilaterally undetectable. Treatment consisted of sodium nitroprusside and heparin. On the third day of the admission, bilateral brachial and femoral artery pulses were lost and his complaints exacerbated. Angiography revealed diffuse vasospasm of the arteries in the both lower extremities (Fig. 1A and C). Because of the lack of response to the ongoing therapy, a single dose of methylprednisolone sodium succinate (1 mg/kg) was given intravenously; the nitroprusside infusion was terminated because of the development of hypotension. The pulses were palpable 2 h after the methylprednisolone dose. Angiography done 12 h after the methylprednisolone dose showed improvement of the vasospasm in the lower extremities. Recovery was uneventful and follow-up evaluation found no abnormalities. DISCUSSION: Although vasodilator agents are first-line therapy in the treatment of ergotism, corticosteroids may be considered as an alternative therapy, especially for intractable cases. The mechanism by which corticosteroids dilate arteries is not clear. CONCLUSIONS: Ischemia in an extremity secondary to ergotamine-induced vasospasm unresponsive to sodium nitroprusside may be treated successfully with methylprednisolone.

Milk and blood ghrelin level in diabetics.

OBJECTIVE: Besides its presence in various tissues, ghrelin has recently been shown to be present in blood and breast milk. No previous studies, however, have evaluated the level of this hormone under the condition of pregestational and gestational diabetes mellitus (P-GDM and GDM, respectively). This study was undertaken to show whether a relation exists between serum and milk ghrelin levels in lactating mothers with and without diabetes. METHODS: Venous blood was obtained from four groups of women (age range 22-37 y): GDM lactating (n = 12), P-GM lactating (n = 3), healthy non-diabetic lactating (n = 14), and healthy non-lactating (n = 14). Colostrum and mature milk samples were collected just before suckling. The ghrelin level was determined by radioimmunoassay and high-performance liquid chromatography. RESULTS: Radioimmunoassay results showed that women with GDM and P-GDM had greater than two-fold lower colostrum and serum levels of ghrelin than did lactating women with no GDM at 2 d after parturition. The GDM and non-diabetic groups at 15 d after delivery, however, showed similar levels of ghrelin in mature milk and serum. High-performance liquid chromatographic results indicated that in serum the deacylated form of ghrelin was 18-fold higher than the acylated form. Furthermore, in milk the acylated form of ghrelin was 24-fold that of the active form. CONCLUSION: These results indicate that mothers with GDM have a substantial (greater than two-fold) decrease in their serum and colostral ghrelin levels. This is, however, a temporary effect lasting only up to early postparturition (2 d after delivery). This peptide hormone restores to completely normal levels at day 15 of parturition, but not P-GDM. The significance of these results in terms of the health of the mother and her newborn, however, has yet to be determined.

Changes in plasma homocysteine levels of rats with experimentally induced hypothyroidism and hyperthyroidism.

INTRODUCTION: It is claimed in a limited number of studies carried out on human beings that plasma homocysteine levels increased in hypothyroid patients and decreased in hyperthyroid patients. OBJECTIVE: The aim of this study is to determine total plasma homocysteine, thyroid function tests, vitamin B12, folic acid and lipid levels and to explore the relations among them in rat models with induced hypothyroidism and hyperthyroidism with a view to investigating whether hypothyroid and hyperthyroid rat models could represent human hypothyroidism and hyperthyroidism models. MATERIAL AND METHOD: The study included 30 male Wistar Albino species rats with a mean weight of 200 - 250 g. Rats were randomly divided into 3 groups as 1) hypothyroid group, 2) hyperthyroid group and 3) control group. Hypothyroidism was induced by adding 10 mg/kg/day propylthiouracil to rats' drinking water for 30 days. In order to induce hyperthyroidism, rats were administered 10 microg/100 g L-thyroxin ampule via intraperitoneal route for 10 days. RESULTS: We found that total plasma homocysteine level of the hypothyroid group was significantly lower than those of the control group (p<0.05) and the hyperthyroid group (p<0.001). Total plasma homocysteine level of the hypothyroid group was found insignificantly higher than that of the control group (p>0.05) and significantly higher than that of the hyperthyroid group (p<0.001). We established a significant and positive correlation between total plasma homocysteine level and thyroid hormone levels. We did not identify a significant relation between total plasma homocysteine level and serum folic acid and serum vitamin B12 levels. CONCLUSION: Our findings are different from the findings reported in human hypothyroidism and hyperthyroidism studies. We believe that hypothyroid and hyperthyroid rat models cannot represent human hypothyroidism and hyperthyroidism models.

Novel role for gabapentin in neuroprotection of central nervous system in streptozotocine-induced diabetic rats.

AIM: To investigate the effect of gabapentin on neural [neuron-specific enolase (NSE)] and glial markers [glial fibrillary acidic protein (GFAP) and S100B] in different brain regions of diabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of streptozotocine (50 mg/kg body weight). Rats in one diabetic group received gabapentin (50 mg.kg(-1).d(-1)) and rats in the other diabetic group received vehicle only for 6 weeks. The levels of GFAP, S100B, and NSE were determined by immunoblotting in the hippocampus, cortex, and cerebellum. Lipid peroxidation (LPO as malondialdehyde+ 4-hydroxyalkenals) and glutathione (GSH) levels were also determined in the same brain parts. RESULTS: Total and degraded GFAP content and S100B protein expression in different areas of brain tissues significantly increased in diabetic rats compared to control rats. Similarly, NSE levels were also significantly elevated in hyperglycemic rats. In addition, there was a significant increase in LPO levels in the diabetic rat brain compared to control rat brains. Pretreatment with gabapentin prevented the upregulation of GFAP, S100B, and NSE in all brain regions of diabetic rats. The level of LPO was reduced, but not completely halted, by treatment with gabapentin. CONCLUSION: These results suggest that diabetes causes glial and neuronal injury, possibly as a result of elevated oxidative stress, and that gabapentin protects neurons and glial cells. Thus, we predict that gabapentin treatment will attenuate the hippo-campal and cortical neurodegeneration observed during diabetes mellitus in rats.