Increased intestinal permeability in patients with short bowel syndrome is not affected by parenteral nutrition.

The aim of our study was to assess the presence and degree of intestinal leakage in subjects suffering from short bowel syndrome (SBS) and its modification by parenteral nutrition. To this end we assessed circulating levels of selected makers of intestinal permeability including zonulin, fatty acid binding protein 2 (FABP-2), citrulline and glucagon-like peptide 2 (GLP-2). We also measured lipopolysaccharide binding protein (LBP) as a marker of circulating levels of lipopolysaccharide acting through the CD14 molecule. Eleven SBS and 10 age- and BMI-matched control subjects were included into the study. The effect of parenteral nutrition was assessed after 14 days, 6 and 12 months from its initiation, respectively. At baseline, SBS patients had increased gut permeability as measured by zonulin (47.24+/-2.14 vs. 39.48+/-1.20 ng/ml, p=0.006) and LBP (30.32+/-13.25 vs. 9.77+/-0.71 microg/ml, p<0.001) compared to healthy controls. Furthermore, SBS subjects had reduced FABP-2, unchanged citrulline and increased sCD14 and GLP-2 relative to control group. Throughout the whole study period the administered parenteral nutrition had no significant effect on any of the studied parameters. Taken together, our data show that patients with short bowel syndrome have increased intestinal permeability that is not affected by parenteral nutrition.

Stable COX17 Downregulation Leads to Alterations in Mitochondrial Ultrastructure, Decreased Copper Content and Impaired Cytochrome c Oxidase Biogenesis in HEK293 Cells.

Cox17 is an assembly factor that participates in early cytochrome c oxidase (COX, CcO) assembly stages. Cox17 shuttles copper ions from the cytosol to the mitochondria and, together with Sco1 and Sco2, provides copper ions to the Cox1 and Cox2 mitochondrially encoded subunits. In Saccharomyces cerevisiae, Cox17 also modulates mitochondrial membrane architecture due to the interaction of Cox17 with proteins of the MICOS complex (mitochondrial contact site and cristae organizing system). There is currently no data regarding the impact of long-term Cox17 deficiency in human cells. Here, we present construction and characterization of three stable COX17 shRNA-downregulated HEK293 cell lines that have less than 10 % of the residual Cox17 protein level. Cox17-depleted cell lines exhibited decreased intramitochondrial copper content, decreased CcO subunit levels (Cox1, Cox4 and Cox5a) and accumulation of CcO subcomplexes. Similarly to yeast cells, mitochondria in Cox17-downregulated HEK293 cell lines exhibited ultrastructural changes including cristae reduction and mitochondrial swelling. Characterization of the molecular pathogenesis of long-term Cox17 deficiency complements our knowledge of the mitochondrial copper metabolism and assembly of cytochrome c oxidase in human cells.

Changes in transcription pattern lead to a marked decrease in COX, CS and SQR activity after the developmental point of the 22(nd) gestational week.

Tissue differentiation and proliferation throughout fetal development interconnect with changes in the oxidative phosphorylation system (OXPHOS) on the cellular level. Reevaluation of the expression data revealed a significant increase in COX4 and MTATP6 liver transcription levels after the 22(nd) gestational week (GW) which inspired us to characterize its functional impact. Specific activities of cytochrome c oxidase (COX), citrate synthase (CS), succinate-coenzyme Q reductase (SQR) and mtDNA determined by spectrophotometry and RT-PCR were studied in a set of 25 liver and 18 skeletal muscle samples at 13(th) to 29(th) GW. Additionally, liver hematopoiesis (LH) was surveyed by light microscopy. The mtDNA content positively correlated with the gestational age only in the liver. The activities of COX, CS and SQR in both liver and muscle isolated mitochondria significantly decreased after the 22(nd) GW in comparison with earlier GW. A continuous decline of LH, not correlating with the documented OXPHOS-specific activities, was observed from the 14(th) to the 24(th) GW indicating their exclusive reflection of liver tissue processes. Two apparently contradictory processes of increasing mtDNA transcription and decreasing OXPHOS-specific activities seem to be indispensable for rapid postnatal adaptation to high energy demands. The inadequate capacity of mitochondrial energy production may be an important factor in the mortality of children born before the critical developmental point of the 22(nd) GW.

Laparoscopic sleeve gastrectomy differentially affects serum concentrations of FGF-19 and FGF-21 in morbidly obese subjects.

OBJECTIVE: Fibroblast growth factor (FGF)-19 and FGF-21 are novel metabolic regulators that improve insulin resistance and obesity in rodents. The aim of the study was to assess the effects of laparoscopic sleeve gastrectomy (LSG) on serum concentrations of FGF-19 and FGF-21 along with circulating bile acids and other relevant hormonal and biochemical parameters. DESIGN AND METHODS: Seventeen females with obesity undergoing LSG and 15 lean healthy females were included into the study. Anthropometric and biochemical parameters, serum concentrations of FGF-19 and -21, insulin, adiponectin, leptin, C-reactive protein, resistin, amylin (total), ghrelin (active), glucagon-like peptide 1 (GLP-1, active), glucose-dependent insulinotropic peptide (GIP, total), peptide YY (PYY, total), pancreatic polypeptide (PP), and bile acids, and mRNA expression of selected adipokines and inflammatory markers in bioptic samples of subcutaneous fat were assessed at baseline and 6, 12, and 24 months after LSG. RESULTS: LSG markedly decreased body weight, BMI, waist circumference, and insulin levels and improved systemic inflammation and lipid levels. FGF-19 concentrations increased and FGF-21 concentrations decreased after LSG along with increased adiponectin and decreased leptin, amylin, and ghrelin levels. GLP-1, GIP, PP, and circulating bile acids were not affected by LSG. PYY decreased significantly 24 months after surgery only. mRNA expression analysis in subcutaneous fat showed markedly reduced proinflammatory state. CONCLUSIONS: Our results indicate that increased FGF-19 and decreased ghrelin concentrations could have partially contributed to the improvement of systemic inflammation and some metabolic parameters after LSG, while changes of FGF-21 are rather secondary because of weight loss.

RAPD and rep-PCR fingerprinting for characterization of Bifidobacterium species.

DNA fingerprinting methods, RAPD with 7 random primers, and rep-PCR using both BOXA1R and (GTG)(5) ones, were used for the discrimination of 16 type and collection Bifidobacterium strains of 9 species of human origin, B. animalis ssp. animalis and B. animalis ssp. lactis and 7 Bifidobacterium strains collected in the Culture Collection of Dairy Microorganisms (CCDM). Both RAPD and rep-PCR methods provided similar results. The strains were identified as B. animalis ssp. lactis (6 strains) and B. adolescentis (1 strain). The reclassification of the collection strain CCM 3761 as B. pseudocatenulatum species (previously classified as B. adolescentis) was confirmed.

Adiponectin and resistin gene polymorphisms in patients with anorexia nervosa and obesity and its influence on metabolic phenotype.

Genes for adiponectin and resistin are candidate genes of insulin resistance and type 2 diabetes mellitus. The aim of our study was to determine the frequency of single nucleotide polymorphisms (SNP) 45T>G and 276G>T of the adiponectin gene and 62G>A and -180C>G of the resistin gene in patients with obesity (OB), anorexia nervosa (AN) and in control healthy normal-weight women (NW) and to study the influence of particular genotypes on serum concentrations of these hormones and on insulin sensitivity. Serum adiponectin, resistin, tumor necrosis factor alpha (TNF-alpha), insulin, cholesterol, glycated hemoglobin (HbA1c) and blood glucose levels were measured in 77 patients with OB, 28 with AN and 38 NW. DNA analysis was carried out by polymerase chain reaction with restriction analysis of PCR product. The presence of SNP ADP+276 G>T allele was accompanied by higher cholesterol levels in AN patients, higher adiponectin concentrations in OB patients and lower HbA1c levels in NW. SNP of the resistin gene 62G>A was associated with lower HbA1c in NW and higher cholesterol concentrations in OB group. The carriers of the minor G allele in the position -180 of the resistin gene within AN group had significantly higher BMI relative to non-carriers. We conclude that polymorphisms in adiponectin and resistin genes can contribute to metabolic phenotype of patients with obesity and anorexia nervosa.

[Polymorphisms of adiponectin and resistin genes in patients with obesity and anorexia nervosa]. / Polymorfismy genu pro adiponectin a rezistin u pacientek s mentální anorexií a obezitou--pilotní studie.

BACKGROUND: Adiponectin and resistin are hormones that may represent a link between obesity and insulin resistance. Genes for these hormones are new candidate genes of insulin resistance and type 2 diabetes mellitus. The aim of our study was to determine the frequency of single nucleotide polymorphisms 45T > G and 276T > G of adiponectin gene and 62G > A and -180C > G of resistin gene in patients with obesity, anorexia nervosa and in lean women and to study the influence of particular genotypes on serum concentrations of these hormones. METHODS AND RESULTS: Serum adiponectin, resistin, TNF-alfa and insulin levels were measured in 51 patients with obesity, 17 with anorexia nervosa and 17 lean women. DNA analysis was carried out by means of polymerase chain reaction (PCR) with restriction analysis of PCR product (RFLP). Adiponectin levels were lowest in obese women and highest in anorexia nervosa patients. Resistin concentrations were lowest in anorexia nervosa and highest in obese patients. Genotype analysis within respective groups showed no differences in assessed parameters when comparing different adiponectin and resistin polymorphisms. The only difference detected was significantly higher BMI in G/G genotype relative to T allele carries in 276 position of ADP gene in control group (23.48 +/- 0.85 vs. 19,7 +/- 0.95, p < 0.05). In anorexia nervosa patients, frequency of G allele in RETN -180 polymorphism was significantly higher relative to control group (p < 0.05). CONCLUSIONS: Polymorphisms 45T > G a 276T > G of ADP gene and 62G>A and -180C > G RETN gene did not influence serum ADP and RETN concentrations. BMI was influenced by T allele presence in 276 position of ADP gene in control group only. Anorexia nervosa patients had higher frequency of G allele of RETN -180 polymorphism compared to healthy women.

Lateralized ictal immobility of the upper limb in patients with temporal lobe epilepsy.

The primary aim of this study was to establish the incidence and the lateralizing value of 'lateralized ictal immobility of the upper limb' (LIL) in patients suffering from temporal lobe epilepsy (TLE), and to describe the connection between LIL and other clinical ictal signs. We retrospectively reviewed video records of 87 patients with TLE. We reviewed a total of 276 focal epileptic seizures with or without secondary generalization. We studied the incidence of LIL, its lateralizing value, and its relationship to other ictal clinical signs. Of the 87 patients, 49 had undergone a successful resective surgery at least 1 year prior to the study. LIL is a late sign in the course of partial seizure. It occurred in 25 of our 87 patients (28.7%), and in 47 of 276 seizures (17.1%). In all of the evaluated seizures, LIL occurred contralateral to the side of seizure onset (P < 0.001). LIL was always associated with ipsilateral upper limb automatisms, and in 63.1% of the occurrences, it was immediately followed by ictal dystonia. LIL is a more accurate term to describe what has previously been called 'ictal paresis' in the literature. Due to the inability to execute proper testing during a partial seizure, it is better to use the term LIL when making a visual analysis of a seizure. LIL is a more suitable term to describe the studied ictal sign. It is a relatively frequent sign in patients with TLE. LIL has an excellent lateralizing value for the contralateral hemisphere. It is a negative motor sign, and its genesis is probably associated with the epileptic involvement of the contralateral frontal lobe.

Short-term very low calorie diet reduces oxidative stress in obese type 2 diabetic patients.

Oxidative stress is higher in obese diabetic than in non-diabetic subjects. This pilot study evaluates oxidative stress during short-term administration of a very low calorie diet in obese persons. Nine obese Type 2 diabetic patients (age 55+/-5 years, BMI 35.9+/-1.9 kg/m2) and nine obese non-diabetic control subjects (age 52+/-6 years, BMI 37.3+/-2.1 kg/m2) were treated by a very low calorie diet (600 kcal daily) during 8 days stay in the hospital. Serum cholesterol, triglycerides, non-esterified fatty acids (NEFA), beta-hydroxybutyrate (B-HB), ascorbic acid (AA), alpha-tocopherol (AT), plasma malondialdehyde (MDA) and superoxide dismutase (SOD) activity in erythrocytes were measured before and on day 3 and 8 of very low calorie diet administration. A decrease of serum cholesterol and triglyceride concentrations on day 8 was associated with a significant increase of NEFA (0.30+/-0.13 vs. 0.47+/-0.11 micromol/l, p<0.001) and B-HB (0.36+/-.13 vs. 2.23+/-1.00 mmol/l, p<0.001) in controls but only of B-HB (1.11+/-0.72 vs. 3.02+/-1.95 mmol/l, p<0.001) in diabetic patients. A significant decrease of plasma MDA and serum AT together with an increase of SOD activity and AA concentration (p<0.01) was observed in control persons, whereas an increase of SOD activity (p<0.01) was only found in diabetic patients after one week of the very low calorie diet. There was a significant correlation between NEFA or B-HB and SOD activity (p<0.01). We conclude that one week of a very low calorie diet administration decreases oxidative stress in obese non-diabetic but only partly in diabetic persons. Diabetes mellitus causes a greater resistance to the effects of a low calorie diet on oxidative stress.

Adipocyte-derived hormones in heroin addicts: the influence of methadone maintenance treatment.

Heroin addiction markedly affects the nutritional and metabolic status and frequently leads to malnutrition. The aim of our study was to compare circulating concentration of adipose tissue-derived hormones leptin, adiponectin and resistin in 12 patients with heroin addiction before and after one-year methadone maintenance treatment with the group of 20 age- and body mass index-matched healthy subjects. Basal serum leptin and adiponectin levels in heroin addicts were significantly decreased (3.4+/-0.4 vs. 4.5+/-0.6 ng/ml and 18.9+/-3.3 vs. 33.9+/-3.1 ng/microl, respectively; p 0.05) while serum resistin concentrations were increased compared to healthy subjects (10.1+/-1.2 vs. 4.6+/-0.3 ng/ml; p 0.05). Moreover, positive correlation of serum leptin levels with body mass index was lost in the addicts in contrast to control group. One year of methadone maintenance treatment normalized serum leptin, but not serum adiponectin and resistin concentrations. In conclusion, circulating concentrations of leptin, adiponectin and resistin are markedly altered in patients with chronic heroin addiction. These alterations appear to be relatively independent of nutritional status and insulin sensitivity.

Plasma ghrelin levels and malnutrition: a comparison of two etiologies.

Ghrelin is a peptide hormone that is involved in regulating growth hormone secretion as well as food intake and energy homeostasis. The aim of this study was to compare changes in plasma ghrelin levels in patients with malnutrition due to anorexia nervosa (AN) or short bowel syndrome (SBS). Blood samples for laboratory analyses were taken from 16 AN patients (plus 13 comparable healthy controls) and 27 SBS patients (plus 13 comparable healthy controls) after an overnight fast. In comparison with their respective control groups, plasma ghrelin levels were increased in the AN patients (p < 0.05) and significantly decreased in the patients with SBS (p < 0.01). These results suggest that quantitative ghrelin secretion in the gut wall is important in determining ghrelin concentrations in the systemic circulation.

[Leptin levels in female patients with restrictive and purgative types of anorexia nervosa]. / Hladiny leptinu u pacientek s restriktivní a purgativní formou mentální anorexie.

BACKGROUND: Anorexia nervosa is an eating disorder with typically chronic course. Two subtypes of anorexia nervosa have been described based on the pattern of eating behavior. Restrictive form of anorexia nervosa is characterized by chronically decreased food intake, while the purgative subtype typically consists of alternating episodes of fasting and overnutrition with factitious vomiting. METHODS AND RESULTS: The aim of this study was to compare anthropometric parameters, serum levels of fat-derived hormone leptin, cholesterol, triacylglycerols and serum leptin/body mass index ratio in patients with restrictive and purgative subtypes of anorexia nervosa respectively. Significantly lower body weight (37.79 +/- 3.93 vs. 49.63 +/- 9.84 kg, p < 0.05), body mass index (13.51 +/- 1.43 vs. 17.75 +/- 2.64 kg/m2, p < 0.05),), body fat percentage (13.28 +/- 2.83 vs. 18.9 +/- 5.65%, p < 0.05), serum leptin (1.117 +/- 0.95 vs. 5.88 +/- 4.7 ng/ml, p < 0.05) and cholesterol levels (4.14 +/- 1.78 vs. 6.31 +/- 1.27 mmol/l, p < 0.05) were found in patient with restrictive relative to purgative subtype of anorexia nervosa. In contrast, no difference in triglyceride levels between both groups was found. Serum leptin levels positively correlated with body mass index and body fat percentage only in patients with purgative subtype of anorexia nervosa (body mass index r = 0.95, p < 0.001, body fat percentage r = 0.64, p < 0.05) but not in those with restrictive subtype. CONCLUSIONS: In conclusion, we demonstrated that serum leptin levels were significantly lower in restrictive relative to purgative subtype of anorexia nervosa. We suggest that this difference is primarily due to distinctions in body fat content.

Soluble leptin receptor levels in patients with chronic renal failure.

Soluble leptin receptor (SLR) is the extracellular part of the leptin receptor. This protein is released into circulation and constitutes the main circulating leptin-binding protein. The aim of our study was to measure SLR concentrations in patients with chronic renal failure (CRF) and healthy subjects and to explore the relationship of SLR to other hormones and cytokines. The patients with CRF had significantly higher serum leptin, TNF-alpha and insulin levels than healthy subjects (25.1+/-23.5 vs. 9.4+/-7.6 ng.ml(-1) (S.D.); 14.2+/-4.2 vs. 4.55+/-2.5 ng.ml(-1); 39.8+/-36.1 vs. 20.3+/-11.1 mU.l(-1)). Serum soluble leptin receptor levels did not differ between these groups (19.1+/-11.3 vs. 19.6+/-6.1 U.ml(-1)). An inverse relationship between serum SLR and leptin levels was found in both groups. In patients with CRF the inverse relationship between SLR and insulin, body fat content and total protein levels were also found, while in healthy subjects only inverse relationship of SLR with insulin and albumin concentrations were detected. We conclude that soluble leptin receptor levels in patients with chronic renal failure do not differ from those of healthy subjects despite higher serum leptin levels in CRF patients. The physiological consequences of this finding require further investigation.

[The changes in serum ghrelin levels and their relationship to IGF-I, its binding proteins and leptin in women patients with anorexia nervosa]. / Zmeny sérových koncentrací ghrelinu a jejich vztah k IGF-I, jeho vazebným proteinum a leptinu u pacientek s mentální anorexií.

Ghrelin is recently discovered peptide hormone involved in the regulation of growth hormone secretion as well as in the regulation of food intake and energetic homeostasis. The study was aimed to describe the changes in ghrelin serum levels in patients with anorexia nervosa and its relationship to some other studied parameters. Sixteen women patients with anorexia nervosa and thirteen healthy women of comparable age were examined clinically and blood samples were taken for estimation of serum levels of ghrelin, leptin, soluble leptin receptor, IGF-I, IGFBP-1 and IGFBP-3. Ghrelin serum levels were significantly increased in the group of patients with anorexia nervosa (p < 0,05). In contrary, serum leptin levels were decreased in the group of patients with anorexia nervosa (p < 0,01). Serum ghrelin levels did not correlate with any other of studied parameters with exception of BMI. We can conclude that serum ghrelin levels are increased in patients with anorexia nervosa and their increase fails to significantly stimulate food intake in this group of patients.

[Erythrocyte membrane sodium transport in patients with type I diabetes mellitus (insulin-dependent diabetes mellitus)]. / Membránový transport sodíku v erytrocytech nemocných s diabetem I. typu (inzulin-dependentní diabetes mellitus).

BACKGROUND: Disorders in sodium metabolism such as an increased total body exchangeable sodium, were found in diabetic patients, although the underlying mechanisms were not clear. The aim of the study was to evaluate red blood cell sodium transport in patients with insulin dependent diabetes mellitus (IDDM) without diabetic nephropathy. METHODS AND RESULTS: Renal hemodynamics using the clearance of inulin and para-amino-hippuric acid during euglycemic clamp and red blood cell sodium transport were examined in 13 IDDM patients without microalbuminuria and in 12 weight-, age- and sex-matched healthy controls. Despite normal renal hemodynamics and intracellular sodium concentrations (6.57 +/- 1.45 vs 5.95 +/- 0.60 mmol/l), IDDM patients had lowered clearance of sodium (2.22 +/_ 1.11 vs 3.24 +/- 1.32 ml/min; p < 0.01) and increased activity of natrium-lithium countertransport compared to C (0.76 +/- 0.50 vs 0.31 +/- 0.22 mmol.l-1 .h-1; p < 0.01). No significant differences between IDDM and C were found in Na+-K+ pump (7.95 +/- 1.95 vs 6.9 +/- 0.99 mmol.l-1 .h-1), in Na+-K+ cotransport (0.68 +/- 0.82 vs 0.82 +/- 0.71 mmol.l-1 .h-1) and in passive Na+ permeability (0.11 +/- 0.05 vs 0.09 +/- 0.02 mmoll.l-1 .h-1). CONCLUSIONS: IDDM patients without signs of diabetic nephropathy have shown changes in sodium-lithium countertransport which could play a role in the pathogenesis of diabetic nephropathy and hypertension in the course of the disease.

Effects of insulin and lipid emulsion on renal haemodynamics and renal sodium handling in IDDM patients.

To evaluate the role of insulin and hypertriglyceridaemia in the regulation of renal haemodynamics and sodium handling in insulin-dependent diabetes mellitus (IDDM), 11 IDDM patients without microalbuminuria and 13 weight-, age-, protein intake- and sex-matched healthy control subjects were studied. Clearances of inulin (Cin), para-amino-hippuric acid (CPAH), sodium (CNa), and lithium (CLi) were measured in four 60-min clearance periods (periods I, II, III and IV) during isoinsulinaemia with lipid emulsion infusion (study 1), a hyperinsulinaemic isoglycaemic clamp with Intralipid infusion (study 2), and during time-controlled isoinsulinaemia (study 3). We found that Cin, CPAH and filtration fraction were comparable in IDDM and control subjects, whereas CNa was decreased in diabetic subjects (2.01 +/- 1.11 vs 3.03 +/- 1.32 ml/min; p < 0.05) due to elevations of proximal tubular fractional and absolute reabsorptions of sodium (p < 0.05). Insulin infusion did not affect Cin, increased CPAH (p < 0.05) and, consequently, lowered the filtration fraction (p < 0.01) in both groups. While acute hyperinsulinaemia resulted in increases in distal tubular fractional and absolute reabsorptions of sodium (p < 0.01) contributing to a fall in CNa (p < 0.01) in control subjects, in diabetic subjects the sodium-retaining effect of insulin was not significant. The lipid emulsion did not alter any of the estimated parameters. We conclude that IDDM without microalbuminuria is associated with a tendency to sodium retention which is not aggravated by insulin when compared to control subjects. Acutely induced hypertriglyceridaemia does not alter renal haemodynamics or renal sodium handling.