Genetic polymorphisms of NOD1 and IL-8, but not polymorphisms of TLR4 genes, are associated with Helicobacter pylori-induced duodenal ulcer and gastritis

BACKGROUND: Intracellular pathogen receptor NOD1 is involved in the epithelial cell sensing Helicobacter pylori, which results in a considerable interleukin (IL)-8 production. The aim of this study was to evaluate the relationship between NOD1 and IL-8 genetic polymorphisms and the development of H. pylori-induced gastritis and duodenal ulcer (DU), as compared with TLR4 polymorphisms. MATERIALS AND METHODS: Eighty-five patients with DU and 135 patients with gastritis were enrolled in the study. Seventy-five serologically H. pylori-positive subjects without gastric or duodenal symptoms served as controls. The G796A (E266K) NOD1 polymorphism was determined by restriction fragment length polymorphism, and the -251 IL-8 polymorphism by amplification refractory mutation system method. The TLR4 (ASP/299/Gly and Thr/399/Ile) gene polymorphisms were examined by melting point analysis. RESULTS: AA homozygote mutant variants of NOD1 were detected in 20% of the H. pylori-positive patients with DU versus 7% of H. pylori-positive patients with gastritis and versus 6% of the H. pylori-positive healthy controls. The IL-8 heterozygote mutant variant was detected with a significantly higher frequency among the DU patients and those with gastritis than among the H. pylori-positive controls. However, no significant correlation concerning the frequency of the TLR4 gene polymorphism could be revealed between any group of patients and the controls. CONCLUSION: E266K CARD4/NOD1, but not the TLR4 gene polymorphism increases the risk of peptic ulceration in H. pylori-positive patients. The -251 IL-8 polymorphism was significantly associated with either gastritis or DU in H. pylori-infected subjects. Host factors including intracellular pathogen receptors and IL-8 production play an important role in H. pylori-induced gastric mucosal damage.     

Design of recombinant antibody microarrays for complex proteome analysis: choice of sample labeling-tag and solid support

Antibody-based microarray is a novel technology with great potential within high-throughput proteomics. The process of designing high-performing antibody (protein) microarrays has, however, turned out to be a challenging process. In this study, we have developed further our human recombinant single-chain variable-fragment (scFv) antibody microarray methodology by addressing two crucial technological issues, choice of sample labeling-tag and solid support. We examined the performance of a range of dyes in a one- or two-color approach on a selection of solid supports providing different surface and coupling chemistries, and surface structures. The set-ups were evaluated in terms of sensitivity, specificity, and selectivity. The results showed that a one-color approach, based on NHS-biotin (or ULS-biotin) labeling, on black polymer Maxisorb slides (or Nexterion slide H) was the superior approach for targeting low-abundant (pg/mL) analytes in nonfractionated, complex proteomes, such as human serum or crude cell supernatants. Notably, microarrays displaying adequate spot morphologies, high S/Ns, minimized nonspecific binding, and most importantly a high selectivity, specificity, and sensitivity (>or=fM range) were obtained. Taken together, we have designed the first generation of a high-performing recombinant scFv antibody microarray technology platform on black polymer Maxisorb slides for sensitive profiling of low-abundant analytes in nonfractionated biotinylated complex proteomes.     

Structural insights into Acyl‐coenzyme A binding domain containing 3 (ACBD3) protein hijacking by picornaviruses

Many picornaviruses hijack the Golgi resident Acyl‐coenzyme A binding domain containing 3 (ACBD3) protein in order to recruit the phosphatidylinositol 4‐kinase B (PI4KB) to viral replication organelles (ROs). PI4KB, once recruited and activated by ACBD3 protein, produces the lipid phosphatidylinositol 4‐phosphate (PI4P), which is a key step in the biogenesis of viral ROs. To do so, picornaviruses use their small nonstructural protein 3A that binds the Golgi dynamics domain of the ACBD3 protein. Here, we present the analysis of the highly flexible ACBD3 proteins and the viral 3A protein in solution using small‐angle X‐ray scattering and computer simulations. Our analysis revealed that both the ACBD3 protein and the 3A:ACBD3 protein complex have an extended and flexible conformation in solution.     

Does food play a prominent role in visual attention to disgusting stimuli?

Journal of Ethology - Disgust is a basic emotion which protects individuals from potential contamination. It is hypothesized that disgust evolved primarily as a mechanism against oral contamination...     

Genetic Differentiation in Anthocyanin Content among Berry Fruits

Anthocyanins are widely distributed secondary metabolites that play an essential role in skin pigmentation of many plant organs and microorganisms. Anthocyanins have been associated with a wide range of biological and pharmacological properties. They are also effective agents in the prevention and treatment of many chronic diseases. Berries are particularly abundant in these compounds; therefore, their dietary intake has health-promoting effects. The aim of this study was to identify and determine the anthocyanin content in selected species and cultivars of berry fruits, such as raspberry, blackberry, red currant, blackcurrant, and highbush blueberry, widely consumed by Europeans. The concentrations of anthocyanins were determined by HPLC, identifying individual compounds: cyanidin-3-O-glucoside, pelargonidin-3-O-glucoside, delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-rutinoside, delphinidin-3-O-galactoside, cyanidin-3-O-galactoside, and malvidin-3-O-galactoside. The experimental data showed that the selected species and cultivars of berry fruits differ significantly in the contents of anthocyanins. Among all species tested, blackberry and blackcurrant were characterized significantly by the highest content of anthocyanins (sum), while the lowest content was found in red currant fruits. Additionally, the content of individual anthocyanin compounds in particular species and cultivars was also different. Considering the high content of anthocyanins and their potential positive impact on human health and protection against disease, berries should be part of healthy nutrition.     

The Role of Vocalization in the Communication between Red Deer Hinds and Calves

Eleven hinds and their calves were observed on a deer farm in order to study their vocal communication. Red deer calves are ‘hiders’, and therefore it was believed that they would need to recognize their mother's voice, but the hinds would not need to recognize the voice of their offspring. The following predictions were tested. First, the calf can recognize its mother by voice. Second, the hind's ability to recognize her calf by voice is less pronounced. The study had two parts. Part I was focused on spontaneous behaviour of the animals. Part II involved experiments with audio recordings. It was observed that the hinds vocalized more frequently than the calves, and it appeared that the vocalization served primarily a contact function. The calves seemed to recognize their mothers' voice better than vice versa. For the hinds, vision was more important than hearing. Thus, the predictions were confirmed.     

The Function of Mounting Behaviour in Farmed Red Deer Calves

The function of mounting behaviour was studied in farmed red deer (Cervus elaphus) calves. On the basis of previous work, we tested two alternative hypotheses about the function of this behaviour. The first hypothesis deals with the proximate function of the behaviour. Three predictions were tested: (1a) mounting behaviour attracts the attention of the mother and/or consolidates the mother–calf bond; (1b) mounting is intended to obtain more milk; (1c) mounting is intended to prevent other calves from sucking from the mother. The second hypothesis deals with an ultimate function in practising for future life with two predictions tested: (2a) mounting behaviour is part of calves' play behaviour promoting development of the calf's locomotor and social skills; (2b) calves gain sexual experience through mounting behaviour. For the study, 50 hind‐calf pairs were observed. The maternity of individual hinds was confirmed by a genetical analysis. The hinds were classified as ‘maternal’ and ‘non‐maternal’ and the calves as ‘filial’ and ‘non‐filial’. We recorded 40 cases of mounting behaviour involving 25 hinds and 21 calves. Our results suggest that the mounting behaviour of red deer calves serves several different functions depending on the circumstances. In association with suckling, calves of both sexes mounted maternal hinds mainly to attract their attention and to achieve another suckling. In situations not associated with suckling, mounting by male calves might be considered part of their sexual training. On the other hand, mounting by female calves probably reflects the attempt to maintain contact with their mother in tense situations.     

Expression, purification and catalytic activity of Lupinus luteus asparagine beta-amidohydrolase and its Escherichia coli homolog.

We describe the expression, purification, and biochemical characterization of two homologous enzymes, with amidohydrolase activities, of plant (Lupinus luteus potassium-independent asparaginase, LlA) and bacterial (Escherichia coli, ybiK/spt/iaaA gene product, EcAIII) origin. Both enzymes were expressed in E. coli cells, with (LlA) or without (EcAIII) a His-tag sequence. The proteins were purified, yielding 6 or 30 mg.L(-1) of culture, respectively. The enzymes are heat-stable up to 60 degrees C and show both isoaspartyl dipeptidase and l-asparaginase activities. Kinetic parameters for both enzymatic reactions have been determined, showing that the isoaspartyl peptidase activity is the dominating one. Despite sequence similarity to aspartylglucosaminidases, no aspartylglucosaminidase activity could be detected. Phylogenetic analysis demonstrated the relationship of these proteins to other asparaginases and aspartylglucosaminidases and suggested their classification as N-terminal nucleophile hydrolases. This is consistent with the observed autocatalytic breakdown of the immature proteins into two subunits, with liberation of an N-terminal threonine as a potential catalytic residue.     

Sclerotia of the acellular (true) slime mould <Emphasis Type="Italic">Fuligo septica</Emphasis> as a model to study melanization and anabiosis

Acellular (true) slime moulds (Myxomycetes) are capable of a transition to the stage of sclerotium — a dormant form of plasmodium produced under unfavourable environmental conditions. In this study, sclerotia of Fuligo septica were analyzed by means of electron paramagnetic resonance (EPR) spectroscopy. The moulds were cultivated in vitro on filter paper, fed with oat flour, and kept until the plasmodia began to produce sclerotia. The obtained sclerotia differed in colour from yellow through orange to dark-brown. The EPR spectra revealed a free radical, melanin-like signal correlated with the depth of the colour; it was strongest in the dark sclerotia. Sclerotization only took place when the plasmodia were starved and very slowly dried. Only the yellow sclerotia were able to regenerate into viable plasmodia. This suggests that myxomycete cytoplasm dehydration is an active process regulated metabolically. Plasmodial sclerotization may therefore serve as a convenient model system to study the regulation of cytoplasmatic water balance, and sclerotia as a convenient material for EPR measurements, combining the quality of plasmodia with the technical simplicity of the measurements characteristic of dry spores. Darkening of the sclerotia is most probably a pathological phenomenon connected with the impairment of water balance during sclerotization. Paper authored by participants of the international conference: XXXIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University, Zakopane, March 7–11, 2007, “The Cell and Its Environment”. Publication costs were partially covered by the organisers of this meeting.