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Unexpected bleeding after Exenatide treatment: a causative relationship or a coincidence?


Objectives. Diabetes mellitus is an endemic disease of the current era. It is important to treat it properly. All antidiabetic medications have side effects and various safety profiles.

Case report. Fifty-two years old patient with type II diabetes mellitus, who had spontaneous cutaneous and intra muscular bleeding after starting treatment with Exenatide. The patient’s history did not include any kind of spontaneous bleeding. Investigations did not reveal abnormal platelets count and function or coagulation profile. The use of the Exenatide was discontinued and during one year of follow-up, the patient did not experience an additional occurrence of spontaneous bleeding.

Conclusions. To the best of our knowledge, this is the first report of spontaneous bleeding probably caused by Exenatide. The exact pathophysiology, by which the drug can cause spontaneous bleeding, is still not clear and has to be revealed.

Open access
c-Fos expression in the hypothalamic paraventricular nucleus after a single treatment with a typical haloperidol and nine atypical antipsychotics: a pilot study


Objective. The aim of the present study was to find out whether acute effect of different doses of selected antipsychotics including aripiprazole (ARI), amisulpride (AMI), asenapine (ASE), haloperidol (HAL), clozapine (CLO), risperidone (RIS), quetiapine (QUE), olanzapine (OLA), ziprasidone (ZIP), and paliperidone (PAL) may have a stimulatory impact on the c-Fos expression in the hypothalamic paraventricular nucleus (PVN) neurons.

Methods. Adult male Wistar rats weighing 280–300 g were used. They were injected intraperitoneally with vehicle or antipsychotics in the following doses (mg/kg of b.w.): ARI (1, 10, 30), AMI (10, 30), ASE (0.3), HAL (1.0, 2.0), CLO (10, 20), RIS (0.5, 2.0), QUE (10, 20), OLA (5, 10), ZIP (10, 30), and PAL (1.0). Ninety min later, the animals were anesthetized with Zoletil and Xylariem and sacrificed by a transcardial perfusion with 60 ml of saline containing 450 μl of heparin (5000 IU/l) followed by 250 ml of fixative containing 4% paraformaldehyde in 0.1 M phosphate buffer (PB, pH 7.4). The brains were postfixed in a fresh fixative overnight, washed two times in 0.1 M PB, infiltrated with 30% sucrose for 2 days at 4 °C, frozen at −80 °C for 120 min, and cut into 30 μm thick serial coronal sections at −16 °C. c-Fos profiles were visualized by nickel intensified DAB immunohistochemistry and examined under Axio-Imager A1 (Zeiss) light microscope.

Results. From ten sorts of antipsychotics tested, only six (ARI-10, CLO-10 and CLO-20, HAL-2, AMI-30, OLA-10, RIS-2 mg/kg b.w.) induced distinct c-Fos expression in the PVN. The antipsychotics predominantly targeted the medial parvocellular subdivision of the PVN.

Conclusions. The present pilot study revealed c-Fos expression increase predominantly in the PVN medial parvocellular subdivision neurons by action of only several sorts of antipsychotics tested indicating that this structure of the brain does not represent a common extra-striatal target area for all antipsychotics.

Open access
Co-exposure to endocrine disruptors: effect of bisphenol A and soy extract on glucose homeostasis and related metabolic disorders in male mice


Objectives. Bisphenol A (BPA) is a xenoestrogen, which is commonly used as a monomer of polycarbonate plastics food containers and epoxy resins. Little is known about the interaction effects between xeno- and phyto- estrogens on glucose homeostasis or other metabolic disorders. The aim of this study was to examine effects of individual or combined exposure to low doses of BPA and soy extract on glucose metabolism in mice with the goal to establish its potential mechanisms.

Methods. Fifty-four male mice were randomly divided into six groups. Mice were treated with soy extract at 60 or 150 mg/kg by daily gavage with or without subcutaneously administration of BPA (100 μg/kg/day) for four weeks at the same time, while the control group received a vehicle. Serum levels of fasting glucose, insulin, adiponectin, testosterone, malondialdehyde (MDA), and total antioxidant capacity (TAC) were measured. Homeostatic model assessment-β cell function (HOMA-β) index was also determined.

Results. BPA exposure induced hyperglycemia and significantly reduced HOMA-β, serum levels of insulin, adiponectin, testosterone, and TAC and noticeably enhanced MDA in BPA group compared to control one. While treatment with soy extract in high dose (150 mg/kg) significantly decreased the levels of fasting blood glucose and MDA and notably improved the serum levels of insulin, HOMA-β, and TAC compared to BPA group.

Conclusion. Soy extract may protect against some adverse effects of BPA. These findings represent the first report suggesting a potential effect between soy extract and BPA in low doses, however, further studies are needed to confirm these results.

Open access
Darapladib inhibits atherosclerosis development in type 2 diabetes mellitus Sprague-Dawley rat model


Objective. Increase in the low-density lipoprotein (LDL) level in diabetes mellitus and atherosclerosis is related to lipoprotein associated phospholipase A2 (Lp-PLA2). Lp-PLA2 is an enzyme that produces lysophosphatidylcholine (LysoPC) and oxidized nonesterified fatty acids (oxNEFA). LysoPC regulates inflammation mediators, including intra-cellular adhesion molecule-1 (ICAM-1). Darapladib is known as a Lp-PLA2 specific inhibitor. The aim of this study was to reveal the effect of darapladib on the foam cell number, inducible nitric oxide synthase (iNOS), and ICAM-1 expression in aorta at early stages of the atherosclerosis in type 2 diabetes mellitus Sprague-Dawley rat model.

Methods. Thirty Sprague-Dawley male rats were divided into 3 main groups: control, rats with type 2 diabetes mellitus (T2DM), and T2DM rats treated with darapladib (T2DM-DP). Each group was divided into 2 subgroups according the time of treatment: 8-week and 16-week treatment group. Fasting blood glucose, insulin resistance, and lipid profile were measured and analyzed to ensure T2DM model. The foam cells number were detected using hematoxylin-eosin (HE) staining and the expression of iNOS and ICAM-1 was analyzed using double immunofluorescence staining.

Results. Induction of T2DM in male Sprague-Dawley rats after high fat diet and streptozotocin injection was confirmed by elevated levels of total cholesterol and LDL and increased fasting glucose and insulin levels compared to controls after both times of treatment. Moreover, T2DM in rats induced a significant increase (p<0.05) in the foam cells number and iNOS and ICAM-1 expression in aorta compared to controls after both treatment times. Darapladib treatment significantly reduced (p<0.05) foam cells number as well as iNOS expression in aorta in rats with T2DM after both treatment times. A significant decrease (p<0.05) in ICAM-1 expression in aorta was observed after darapladib treatment in rats with T2DM only after 8 weeks of treatment.

Conclusion. Our data indicate that darapladib can decrease the foam cells number, iNOS, and ICAM-1 expression in aorta at the early stages of atherosclerosis in T2DM rat model.

Open access
Glucometabolic effects of single and repeated exposure to forced-swimming stressor in Sprague-Dawley rats


Objectives. We aimed to evaluate the effects of a single (acute) and repeated (chronic) exposure to forced-swimming stressor on glucose tolerance, insulin sensitivity, lipid profile and glycogen content in male rats.

Methods. Thirty adult male Sprague-Dawley rats (12 weeks old) were divided randomly into five groups: control group, single exposure (SE) to forced-swim stressor, repeated exposure to forced-swim stressor for 7 days (RE7), 14 days (RE14) and 28 days (RE28). Glucose tolerance test and Homeostatic Model Assessment-Insulin Resistance (HOMA-IR) were undertaken on fasting rats to obtain glucose and insulin profiles. ELISA was performed to assess plasma insulin and corticosterone levels. Total cholesterol, triglyceride, high- and low-density lipoproteins, hepatic and skeletal glycogen content were also determined.

Results. Repeated exposure to stressor induced glucose intolerance and insulin resistance in the experimental rats. Results showed that all RE groups exhibited a significantly higher area under the curve compared with others (p=0.0001); similarly, HOMA-IR increased (p=0.0001) in all RE groups compared with control. Prolonged exposure to stressor significantly increased the plasma insulin and corticosterone levels but decreased the glycogen content in the liver and skeletal muscle when compared with the control group. Additionally, chronic stressor significantly increased the total cholesterol and triglyceride levels, however, acute stressor produced significantly elevated high-density lipoproteins level.

Conclusions. In conclusion, repeated exposure to forced-swimming stressor induced glucose intolerance and insulin resistance in rats by disrupting the insulin sensitivity as well as heightening the glycogenolysis in the liver and skeletal muscle. Acute stressor was unable to cause glucose intolerance and insulin resistance but it appears that may have a positive effect on the lipid metabolism.

Open access
Mutations in SURF1 are important genetic causes of Leigh syndrome in Slovak patients


Objectives. Leigh syndrome is a progressive early onset neurodegenerative disease typically presenting with psychomotor regression, signs of brainstem and/or basal ganglia disease, lactic acidosis, and characteristic magnetic resonance imaging findings. At molecular level, deficiency of respiratory complexes and/or pyruvate dehydrogenase complex is usually observed. Nuclear gene SURF1 encodes an assembly factor for cytochrome c-oxidase complex of the respiratory chain and autosomal recessive mutations in SURF1 are one of the most frequent causes of cytochrome c-oxidase-related Leigh syndrome cases. Here, we aimed to elucidate the genetic basis of Leigh syndrome in three Slovak families.

Methods and results. Three probands presenting with Leigh syndrome were selected for DNA analysis. The first proband, presenting with atypical LS onset without abnormal basal ganglia magnetic resonance imaging findings, was analyzed with whole exome sequencing. In the two remaining probands, SURF1 was screened by Sanger sequencing. Four different heterozygous mutations were identified in SURF1: c.312_321delinsAT:p.(Pro104Profs*1), c.588+1G>A, c.823_833+7del:p. (?) and c.845_846del:p.(Ser282Cysfs*9). All the mutations are predicted to have a loss-of-function effect.

Conclusions. We identified disease-causing mutations in all three probands, which points to the important role of SURF1 gene in etiology of Leigh syndrome in Slovakia. Our data showed that patients with atypical Leigh syndrome phenotype without lesions in basal ganglia may benefit from the whole exome sequencing method. In the case of probands presenting the typical phenotype, Sanger sequencing of the SURF1 gene seems to be an effective method of DNA analysis.

Open access
Possible association of ghrelin/obestatin balance with cardiometabolic risk in obese subjects with Helicobacter pylori


Objectives. Helicobacter pylori (H. pylori) is a common gastric infection associated with extragastric conditions. The association between H. pylori infection and obesity is unclear. H. pylori may affect gut hormones involved in food intake and energy expenditure. The aim of this study is to evaluate ghrelin/obestatin balance and leptin in obese subjects with H. pylori infection.

Methods. Sixty healthy volunteers were divided into: obese and non-obese groups. Each group was divided into H. Pylori positive or H. pylori negative. Anthropometric parameters, H. pylori status, serum glucose, insulin level, and lipid profile were estimated with calculation of Homeostasis Model Assessment Insulin Resistance (HOMA-IR). Serum levels of ghrelin, obestatin, and leptin were evaluated.

Results. Significant increase was found in serum glucose, insulin and HOMA-IR ratio in obese subjects with positive H. pylori as compared to other groups. H. pylori positive obese subjects showed significantly increased ghrelin, ghrelin/obestatin balance, and leptin with a significant decrease in obestatin as compared to negative subjects. Ghrelin/obestatin ratio positively correlated with weight, body mass index, waist, glucose, insulin, HOMA-IR, leptin, cholesterol, triglycerides, low density cholesterol and also with H. pylori antigen in the same group.

Conclusions. It can be concluded that ghrelin, obestatin, and leptin are affected by presence of H. pylori seropositivity in obese subjects. The higher ghrelin levels and ghrelin/obestatin ratio with lowered obestatin could be considered as a gastro-protective effect against inflammation induced by H. pylori.

Open access
Possible protective effect of curcumin on the thyroid gland changes induced by sodium fluoride in albino rats: light and electron microscopic study


Objectives. Thyroid gland regulates the body’s metabolic rate and plays an exquisitely important role in the human health. Fluoride exposure can affect thyroid function. Curcumin is a potent antioxidant that works through several mechanisms. The aim of the present study was to demonstrate the hormonal, histological, and ultrastructural changes occurred in the thyroid gland induced by exposure to sodium fluoride (NaF) and study the possible protective effect of curcumin on the NaF-induced effects.

Methods. Thirty male albino rats were randomly divided into 3 equal groups (10 rats each): the control group, NaF group, and NaF+Curcumin (NaF+Cur) group. Thyroid-stimulating hormone (TSH), triiodothyronine (T3) and thyroxine (T4) levels were assayed and thyroid tissues processed for light and transmission electron microscopic study.

Results. In NaF group, serum T3 and T4 levels were significantly decreased whereas TSH level was significantly increased compared to the control group. Thyroid tissues showed flattening of the epithelial lining with several follicular cell degenerations, hyperplasia, decreased colloid, disrupted basement membrane, cytoplasmic vacuolations, degenerated mitochondria, widening of rough endoplasmic reticulum cisternae, and vascular congestion compared to the control group. In the NaF+Cur group, serum TSH levels were significantly decreased in comparison with NaF group and no significant difference in comparison with the control group. Thyroid sections appeared apparently normal compared to the control group and NaF group.

Conclusions. Sodium fluoride affected both the function and structure of the thyroid gland while curcumin was protective against these toxic effects.

Open access
Anthelmintic Resistance and Associated Management Practices in Local Horses in Sokoto Metropolis, Nigeria


This study was carried out to assess the management practices used in the control of gastrointestinal (GI) nematodes of horses and to determine the efficacy of three anthelmintics commonly used in Sokoto metropolis. A questionnaire was administered on management practices, while an anthelmintic efficacy test was carried out using 15 horses. The 15 horses were divided into three groups (A, B and C) comprising of 5 each and treated with albendazole, ivermectin and fenbendazole, respectively. The faecal egg count reduction test (FECRT) was used to determine the efficacy and faecal culture was used to determine the parasite species. Majority of the respondents (80%) claimed to have worm control strategies, but only 32.5% used anthelmintics for the control of GI parasites. 62.5% of respondents designed their deworming plan, while only 25% relied on veterinarians. Most of the treatments were done by the horse owners and/or handlers and they largely depended on visual judgement in dosage determination. Their selection of anthelmintics was based on familiarity and 52.5% of the respondents dewormed their horses six times a year using a particular class of anthelmintic or herbal remedies. Resistance against albendazole as well as suspected resistance against fenbendazole by the GI nematodes identified was observed, while ivermectin demonstrated high efficacy against all nematodes isolated. In conclusion, a single dose of subcutaneous injection of ivermectin was highly effective against gastrointestinal parasites in horses, while the worm control strategies employed by respondents enhanced the selection of nematode resistance to albendazole and fenbendazole.

Open access
Determination of Endoparasites by Faecal Examination in the Wild Boar Population in Vojvodina (Serbia)


The aim of our study was to determine whether and to what extent certain species of helminths and protozoa are present in the wild boar population living in hunting grounds in Vojvodina. For this purpose, 52 faecal samples of hunted wild boars (aged 6 months to 2 years) were examined. Examination of the faeces was performed using classical coproscopic laboratory methods. The following parasite species were identified: Metastrongylus spp. Ascaris suum, Trichuris suis, Physocephalus sexalatus, Strongyloides ransomi, Oesophagostomum sp. / Globocephalus sp., Hyostrongylus rubidus, Gnathostoma hispidum, Eimeria deblecki and Eimeria suis. The obtained results from this study indicated that wild boars are a potential reservoir of a variety of endoparasites, thus endangering the surrounding ecosystem.

Open access