-295 T-to-C promoter region IL-16 gene polymorphism is associated with Whipple's disease.

Whipple's disease (WD) is a rare systemic condition caused, in genetically predisposed subjects, by Tropheryma whipplei, a common bacterium widespread in the environment. The relevance of genetic predisposition in WD is shown by the association with HLA alleles DRB1*13 and DQB1*06 and by the demonstration that, in patients with WD, the cytokine genetic profile is skewed toward a Th2 and Treg response. Since IL-16 is involved in hampering the development of a protective macrophagic response against Tropheryma whipplei, we investigated whether the genetic background of IL-16 is different between patients with WD and controls. The -295 T-to-C polymorphism of the promoter region of the IL-16 gene was studied in 90 patients with WD and 152 healthy controls. Levels of serum IL-16 protein were also tested. The frequency of the wild type T allele was significantly higher in patients with WD compared to the controls (155/180 vs. 235/304; p = 0.02 for the Chi(2) test), odds ratio 1.82 [95 % confidence interval (CI) 1.07-3.10]. The TT genotype was found in 65/90 patients with WD and 88/152 controls (p = 0.026). No relationship was found between serum levels of IL-16 and genotypes. Although the functional consequences of this genetic background on levels of IL-16 and on the course of the disease are still unknown, we found, for the first time, that the wild type T allele and the TT genotype of the -295 polymorphism are associated with WD.

Cytokine genetic profile in Whipple's disease.

Whipple's disease (WD) is a very rare chronic systemic condition characterised by a Th2/T regulatory (Treg) dysregulated immune response versus Tropheryma whipplei, a bacterium widely diffuse in the environment. To investigate whether this Th2/Treg polarised response has a genetic background, we investigated the Th1, Th2, Th17 and Treg cytokine genetic profile of 133 patients with WD. Thanks to the European Consortium on WD (QLG1-CT-2002-01049), the polymorphism of 13 cytokine genes was analysed in 111 German and 22 Italian patients using the polymerase chain reaction with sequence-specific primers (PCR-SSP) technique. The frequencies of the genotypes, haplotypes and functional phenotypes were compared with those obtained in 201 German and 140 Italian controls. Clinical heterogeneity was also considered. Functionally, WD patients may be considered as low producers of TGF-ß1, having an increased frequency of the genotype TGF-ß1+869C/C,+915C/C [12.3 % vs. 3.81 %, odds ratio (OR) = 4.131, p = 0.0002] and high secretors of IL-4, carrying the genotype IL-4-590T/T (5.34 % vs. 1.17 %, OR = 5.09, p = 0.0096). No significant association was found between cytokine polymorphism and clinical variability. Analogously to the recent cellular findings of a Th2/Treg polarised response, we showed that the cytokine genetic profile of WD patients is skewed toward a Th2 and Treg response. This was similar in both German and Italian populations. However, the significant deviations versus the controls are poorer than that expected on the basis of these recent cellular findings.

Metabolism and potency of xenin and of its reduced hexapseudopeptide psi fragment in the dog.

Xenin is a 25 amino acid peptide hormone, secreted into the circulation by specific endocrine cells in the duodenal mucosa. Plasma concentrations are elevated after sham feeding and feeding. In the present study the metabolism of xenin and of a C-terminal fragment was investigated. Xenin, its C-terminal hexapeptide, and a pseudopeptide analog psi (CH2NH) hexapeptide in which a psi reduced bond is introduced in the biologically important dibasic motif of the C-terminus were infused or injected intravenously in 14 anaesthetized dogs. Plasma disappearance time, metabolic clearance rate, the generation of metabolites, and biological effects on exocrine pancreatic secretion were determined employing radioimmunoassay, high pressure liquid chromatography, mass spectrometry (MALDI-MS), and sequence analysis. Half time after steady state infusion of xenin was 3.1 min(-1), that of psi xenin 6 was 6.2(-1) (p<0.01) Plasma concentrations of psi xenin 6 were significantly elevated (p<0.01), pancreatic secretion of volume was augmented by a factor of 50, and output of protein by a factor of 30 compared to unmodified xenin 6. MALDI-MS and sequencing after infusion of xenin revealed a C-terminal octapeptide fragment as primary metabolite. Introduction of a reduced pseudobond in the dibasic motif of xenin dramatically enhances biological potency. This indicates that such a reduced pseudopeptide may be useful in the treatment of bowel paralysis.

Phase III of the migrating motor complex: associated with endogenous xenin plasma peaks and induced by exogenous xenin.

Xenin, a recently discovered peptide produced by specific endocrine cells of the duodenal mucosa, has shown exocrine, endocrine and motility effects in the gastroenteropancreatic system in animal experiments. The aim of the present investigation was to study the role of xenin in the regulation of duodenojejunal motility of humans. Twenty-nine healthy volunteers from the hospital staff gave informed consent to participate in this investigation. In 20 volunteers, we determined plasma concentrations of immunoreactive xenin at 15 min intervals over a mean time period of 8 h fasting and recorded the interdigestive motor activity of the duodenojejunum. In a double-blind randomized crossover study on other nine subjects, synthetic xenin in a dose of 4 pmol kg-1 min-1 or placebo was infused for 10 min intravenously in the interdigestive period and postprandially after a liquid meal. Duodenojejunal motility was recorded simultaneously. Predefined interdigestive xenin plasma peaks were found to be significantly associated with the phases III of the migrating motor complex. In the interdigestive period, xenin induced a premature phase III activity in each volunteer; this was followed by a second phase III in five out of nine subjects. In the postprandial state, xenin significantly increased contraction frequency and the percentage of aborally propagated contractions. These findings suggest a role of the peptide hormone xenin in modulating interdigestive and postprandial duodenojejunal motility in humans.

Localization of xenin-immunoreactive cells in the duodenal mucosa of humans and various mammals.

Xenin is a 25-amino-acid peptide extractable from mammalian tissue. This peptide is biologically active. It stimulates exocrine pancreatic secretion and intestinal motility and inhibits gastric secretion of acid and food intake. Xenin circulates in the human plasma after meals. In this study, the cellular origin of xenin in the gastro-entero-pancreatic system of humans, Rhesus monkeys, and dogs was investigated by immunohistochemistry and immunoelectron microscopy. Sequence-specific antibodies against xenin detected specific endocrine cells in the duodenal and jejunal mucosa of all three species. These xenin-immunoreactive cells were distinct from enterochromaffin, somatostatin, motilin, cholecystokinin, neurotensin, and secretin cells, and comprised 8.8% of the chromogranin A-positive cells in the dog duodenum and 4.6% of the chromogranin A-positive cells in human duodenum. In all three species, co-localization of xenin was found with a subpopulation of gastric inhibitory polypeptide (GIP)-immunoreactive cells. Immunoelectron microscopy in the canine duodenal mucosa demonstrated accumulation of gold particles in round, homogeneous, and osmiophilic secretory granules with a closely adhering membrane of 187 +/- 19 nm diameter (mean +/- SEM). This cell type was found to be identical to the previously described canine GIP cell. Immunocytochemical expression of the peptide xenin in a subpopulation of chromogranin A-positive cells as well as the localization of xenin immunoreactivity in ultrastructurally characterized secretory granules permitted the identification of a novel endocrine cell type as the cellular source of circulating xenin.

Lymphocytic colitis, induced by ticlopidine.

Lymphocytic colitis is a chronic inflammatory colonic disease characterized by watery diarrhea and a dense infiltration of the colonic mucosa with lymphocytes. The etiology is unknown but an immune reaction to various immunostimulatory agents including pathogenic or commensal bacteria, products of bacterial metabolism of dietary degradation, or antigens derived directly from the diet, and autoimmune phenomena are discussed. We observed a patient with all features of lymphocytic colitis characterized by a prominent intraepithelial T-cell component. The colitis resolved completely when therapy with ticlopidine--an agent inhibiting platelet aggregation--was stopped. This observation suggests that medical history concerning drug ingestion may reveal the etiology of lymphocytic colitis and allows cure of this otherwise difficult to treat disorder.

Recurring myocardial infarction in a 35 year old woman.

A 35 year old woman presented with acute myocardial infarction without any of the usual risk factors: she had never smoked; she had normal blood pressure; she did not have diabetes; plasma concentrations of total cholesterol and high and low density lipoprotein cholesterol, fibrinogen, homocysteine, and Lp(a) lipoprotein were normal. She was not taking oral contraceptives or any other medication. Coronary angiography showed occlusion of the left anterior descending coronary artery but no evidence of arteriosclerosis. Medical history disclosed a previous leg vein thrombosis with pulmonary embolism. Coagulation analysis revealed protein C deficiency. The recognition of protein C deficiency as a risk factor for myocardial infarction is important as anticoagulation prevents further thrombotic events, whereas inhibitors of platelet aggregation are ineffective.

Effect of omeprazole treatment on plasma concentrations of the gastric peptides, xenin, gastrin and somatostatin, and of pepsinogen.

The peptide xenin 25 is a gastric mucosal constituent like gastrin, somatostatin and pepsinogen. Gastrin and pepsinogen plasma concentrations increase when the secretion of gastric acid is reduced by proton pump inhibitors. In the present investigation, treatment with omeprazole led to an increase in fasting and postprandial plasma concentrations of xenin, gastrin and pepsinogens A and C (P < 0.05, in each instance), whereas somatostatin plasma levels remained unchanged. Because subcutaneous injection of pentagastrin did not raise xenin plasma concentrations, a direct effect of gastrin on xenin production seems unlikely. This study indicates that xenin plasma concentrations are regulated by intragastric pH, as are those of gastrin and pepsinogen.

Xenin--a review.

Xenin, a 25 amino acid peptide, has been identified in human gastric mucosa in the search for a counterpart to the amphibian octapeptide xenopsin. Xenin is structurally related also to the hypothalamic and ileal peptide neurotensin and is, therefore, a member of the xenopsin/neurotensin/xenin peptide family. The biological activities of these peptides are similar: Xenin has been shown to inhibit pentagastrin-stimulated secretion of acid, to induce exocrine pancreatic secretion and to affect small and large intestinal motility. In the gut, xenin interacts with the neurotensin receptor. Radioimmunoassay and chromatography of postprandial plasma in humans indicate the release of xenin into the circulation. The identification of a 35-amino acid precursor peptide of xenin - proxenin, and a review of the Gen-bank revealed that xenin represents the N terminus of a cytosolic coat protein (alpha-COP) from which xenin can be cleaved by aspartic proteinases such as pepsin and cathepsin E. The physiological role of the peptide xenin is not known.

Fatal pulmonary infection with Mycobacterium celatum in an apparently immunocompetent patient.

Mycobacterium celatum is a recently described mycobacterium isolated from patients who have suppressed cell-mediated immunity, such as AIDS. We present here, to our knowledge, the first report of a fatal pulmonary infection caused by M. celatum in a 73-year-old immunocompetent female patient. The mycobacterium was identified by a 16S rRNA sequence analysis.

Gastroenteropancreatic effects of xenin in the dog.

Xenin is a 25 amino acid peptide detected in the gastric mucosa of various mammals. It has since been found in low concentrations in other tissues. Xenin plasma concentrations increase after meals. The present study reports some gastroenteropancreatic effects of this peptide in the dog. Intravenous infusion of 64 pmol/kg min synthetic xenin significantly inhibited pentagastrin-stimulated gastric acid secretion and stimulated exocrine pancreatic secretion of volume and protein. Further, intravenous infusion of xenin in a dose of 1.0 pmol/kg min stimulated jejunal motility in the anaesthetized dog. An intravenous infusion of 32 pmol/kg min xenin raised plasma concentrations of pancreatic polypeptide, vasoactive intestinal polypeptide, insulin and glucagon. The present experiments therefore indicate manifold bioactive properties of intravenously infused xenin in the dog, with jejunal motility the most sensitive target. Conclusions as to the physiological role of xenin cannot be drawn from the present experiments. The release of other hormonal peptides indicates a complex action of xenin.

Relaxant effect of xenin on rat ileum is mediated by apamin-sensitive neurotensin-type receptors.

The action of xenin, a novel 25-residue peptide of the neurotensin (NT)/xenopsin family, was investigated in isolated rat ileal muscle strips and in dispersed longitudinal smooth muscle cells of rat small intestine in vitro. Xenin relaxes KCl-precontracted ileal strips dose dependently (1 nM-3 microM). The order of potency of the investigated peptides was as follows: xenopsin = NT = xenin > neuromedin N. Kinetensin was inactive. Tetrodotoxin, hexamethonium, tetraethylammonium, 4-aminopyridine, and NG-nitro-L-arginine did not influence the relaxant effects of xenin or NT, whereas the K+ channel blocker apamin nearly abolished their effects. Desensitization against one of the peptides or blockade of NT receptors by SR-48692 prevented the effect of xenin and NT. Structure-activity experiments revealed that the COOH-terminal part of the molecules of xenin and NT is essential for biological activity. Experiments with isolated dispersed smooth muscle cells and binding studies on intestinal smooth muscle cell membranes confirmed and extended the results obtained with muscle strips. In conclusion, xenin relaxes rat ileal smooth muscle via a muscular NT-type apamin-sensitive receptor.

Neurokinetic and myokinetic effects of the peptide xenin on the motility of the small and large intestine of guinea pig.

The effects of xenin 25, a peptide of the neurotensin/xenopsin family, were examined on the motility of the guinea pig jejunum and colon in vitro. In the jejunum, xenin induced a biphasic response: first a small relaxation and then a large contraction. In the colon, xenin induced relaxation. The tenia coli was contracted. Deletion or amidation of the C-terminal leucine inactivated xenin. A peptide sequence of 16 C-terminal amino acids was necessary to elicit a full response in the jejunum, whereas in the colon, the potencies of all fragments of xenin 25, including the 6 C-terminal amino acid sequence (xenin 6), were not significantly different from that of xenin 25. In the jejunum, contraction was abolished or reduced by tetrodotoxin, by atropine, by met-enkephalin, by antagonists to the tachykinin receptors NK1 and NK2 and by the P2-purinoceptor antagonist suramin. L-NNA, phentolamine, methysergide, hexamethonium and apamin had no effect. In the colon, all actions of xenin were tetrodotoxin-resistant; the potency of xenin to relax was reduced by apamin and suramin. The actions of xenin on small and large bowel were attenuated by the neurotensin receptor antagonist SR 48692. The xenin analog neurotensin was significantly less potent than xenin 25 in contracting the jejunum and more potent than xenin 25 in relaxing the colon. We conclude that xenin exerts excitatory effects on a neuronal subtype receptor in the jejunum, with participation of muscarinic, purinergic and tachykinin-related mechanisms. Xenin exerts relaxing effects on the colon by interaction with a myokinetic subtype receptor involving Ca(++)-dependent K+ channels and the P2-purinoceptor.

[Chronic transfusion hepatitis--an underestimated disease]. / Die chronische Transfusionshepatitis--eine unterbewertete Krankheit.

BACKGROUND: The 12th Deutsche Bundestag constituted a fact-finding committee to investigate infections transmitted by blood and blood products. 1,358 patients, infected with HIV, mostly suffering from hemophilia were registered. Transfusion-associated hepatitis was not recorded. METHODS AND RESULTS: In a general hospital, a prospective study during 12 months revealed that chronic hepatitis C was associated in 43% with previous transfusion of blood whereas no case of HIV-infection, transmitted by blood or blood products was observed (p < 0.01). The frequency of blood transfusion in patients without chronic hepatitis and without AIDS was 18.5%, also significantly different from patients with chronic hepatitis C (p < 0.025). CONCLUSION: Frequency and impact of chronic transfusion hepatitis C is underestimated.

Identification of proxenin as a precursor of the peptide xenin with sequence homology to yeast and mammalian coat protein alpha.

Proxenin a precursor of the bioactive peptide xenin, was isolated from canine pancreas by HPLC and identified by mass spectrometry and sequence analysis as a pentatriacontapeptide with a molecular weight of 4035: Met Leu-Thr Lys-Phe-Glu-Thr-Lys-Ser-Ala-Arg-Val-Lys-Gly-Leu-Ser- Phe-His-Pro-Lys-Arg-Pro-Trp.Ile-Leu-Thr-Ser-Leu-His-Asn-Gly-Val-Ile-Glo- Leu-OH. Treatment with pepsin cleaved off 10 C-terminal amino acids and released xenin. Data base search showed amino acid sequence homology of xenin and proxenin with the sequence of coal protein alpha of yeast (62%) and humans (100%). Concentration of the coatomer complex from rabbit liver led to an equimolar enrichment of extractable proxenin. We conclude, therefore, that xenin and proxenin are peptide sequences highly conserved during evolution within the alpha-subunit of the coatomer.