Host response to the presence of Helicobacter spp. DNA in the liver of patients with chronic liver diseases

Literature data indicate an association between the presence of Helicobacter spp. in the liver and the development of hepatocellular carcinoma (HCC). However, the role of H. pylori infections in chronic liver diseases (CLD) remains controversial. The aim of this study was to detect Helicobacter spp. DNA in patients with CLD, and to investigate the host response to the presence of the bacterium in the liver. Helicobacter spp. DNA was detected in 59% samples. H.pylori was the most prevalent species (94%). We estimated the expression level of IL-1 and IL-8 genes. The presence of Helicobacter spp. did not have a significant effect on the gene expression of IL-8 and IL-1.     

Cytokine production using membrane adsorbers: Human basic fibroblast growth factor produced by Escherichia coli

Basic fibroblast growth factor (FGF‐2) is a multifunctional cytokine that regulates various cellular processes both in vitro and in vivo. FGF‐2 is extensively used in embryonic stem cell cultures since it can maintain the cells in an undifferentiated state. However, the high price of FGF‐2 has limited its application in stem cell research. Here we present a fast and efficient process for the purification of FGF‐2 from recombinant Escherichia coli cultures using reusable membrane adsorbers. A high expression level of FGF‐2 (42 mg/g dry cell) was achieved by fed‐batch cultivation of E. coli BL21(DE3). A new combination of cation exchange membrane chromatography and heparin‐sepharose affinity chromatography was used for the purification of the protein. A novel anion exchange membrane chromatography was used in the polishing step to remove endotoxins and DNA. In this new process, about 200 mg soluble FGF‐2 was yielded from 1.9 L culture broth with a purity of 98%. The purified protein was identified to be endotoxin‐free and bioactive. It was successfully tested to keep primate embryonic stem cell and human‐induced pluripotent stem cell pluripotent. Our approach, in which a controlled cultivation process is combined with an optimized fast and versatile downstreaming process, is suitable for low‐cost preparation of bioactive FGF‐2 at bench‐scale and may be beneficial to the effective production of other cytokines.     

Response to: “Response to Different measures of 'genome-wide' DNA methylation exhibit unique properties in placental and somatic tissues

We are responding to a Letter to the Editor addressing the Method section of our paper “Different measures of ‘genome-wide’ DNA methylation exhibit unique properties in placental and somatic tissues.” The letter raised concerns that the protocol for Epigentek’s MethylFlash kit was followed incorrectly based on the wording of an online publication of our article. We admittedly made an error in the language used to describe the MethylFlash protocol in our initial submission and thus this was corrected as soon as it was brought to our attention. However, the error was only in language and not procedure. We are confident that the protocol was followed as stated in the insert provided with the MethylFlash^TM Methylated DNA Quantification kit (Colorimetric).We are responding to a Letter to the Editor addressing the Method section of our paper “Different measures of ‘genome-wide’ DNA methylation exhibit unique properties in placental and somatic tissues” (Price ME, Cotton AM, PeÒaherrera MS, McFadden DE, Kobor MS, Robinson WP. Different measures of “genome-wide” DNA methylation exhibit unique properties in placental and somatic tissues. Epigenetics 2012; 7: 652–63; PMID: 22531475; 10.4161/epi.20221). The letter raised concerns that the protocol for Epigentek’s MethylFlash kit was followed incorrectly based on the wording of an online publication of our article. We admittedly made an error in the language used to describe the MethylFlash protocol in our initial submission and thus this was corrected as soon as it was brought to our attention. However, the error was only in language and not procedure. We are confident that the protocol was followed as stated in the insert provided with the MethylFlash^TM Methylated DNA Quantification kit (Colorimetric).     

Nematodes from the Strait of Magellan and the Beagle Channel (Chile): the genera Cervonema and Laimella (Comesomatidae: Nematoda)

Five species of Cervonema and four species of Laimella are described from the Strait of Magellan and the Beagle Channel, Chile, six species of which are new to science. Cervonema chilensisn. sp. and Cervonema hermanin. sp. are separated from other known species of Cervonema by a short cervical region (less than one head diameter from the front end to the anterior border of the amphids). Cervonema chilensisn. sp. is characterised by a tail length of 5 anal diameters with posterior half filiform; Cervonema hermani n. sp. is characterised by a tail length of 6–9 anal diameter and posterior part (75%) cylindrical. Cervonema shiaen. sp. is characterised by the cephalic seta 4 m long, amphids 9–10 m in diameter; spicules 16 m long and 0.8–0.9 abd; tail 4.7–5.4 anal diameter and 50% posterior part filiform; 4–5 minute precloacal supplements. Laimella subterminatan. sp. is characterised by the subterminal position of the buccal cavity which separates it from the other species of the genus. Laimella annaen. sp. is characterised by the head diameter 9–11 m, cephalic setae and external labial setae 9 + 5 m long, respectively, amphids 7 m in diameter; spicules 28–30 m long; tail 14–17 anal diameter and posterior part (75%) filiform; 5 precloacal supplements. Laimella sandraen. sp. is very close to Laimella annaen.sp. in having similar cephalic sensilla, amphids and spicules. Laimella sandraen. sp., however, can be separated from L.annaen. sp. by the shape of head and the structure of sperm cells, the total body length and the cylindrical part of tail. Cervonema papillatum Jensen, 1988, C. tenuicauda (Stekhoven, 1950) and L. longicauda Cobb, 1920 are found in this area as well. The key of all known species of Cervonemaand Laimellais presented.     

Reversible two-step unfolding of heme–human serum albumin: a 1H-NMR relaxometric and circular dichroism study

Human serum albumin (HSA) participates in heme scavenging, the bound heme turning out to be a reactivity center and a powerful spectroscopic probe. Here, the reversible unfolding of heme–HSA has been investigated by ¹H-NMR relaxometry, circular dichroism, and absorption spectroscopy. In the presence of 6 equiv of myristate (thus fully saturating all available fatty acid binding sites in serum heme–albumin), 1.0 M guanidinium chloride induces some unfolding of heme–HSA, leading to the formation of a folding intermediate; this species is characterized by increased relaxivity and enhanced dichroism signal in the Soret region, suggesting a more compact heme pocket conformation. Heme binds to the folding intermediate with K _d = (1.2 ± 0.1) × 10⁻⁶ M. In the absence of myristate, the conformation of the folding intermediate state is destabilized and heme binding is weakened [K _d = (3.4 ± 0.1) × 10⁻⁵ M]. Further addition of guanidinium chloride (up to 5 M) brings about the usual denaturation process. In conclusion, myristate protects HSA from unfolding, stabilizing a folding intermediate state in equilibrium with the native and the fully unfolded protein, envisaging a two-step unfolding pathway for heme–HSA in the presence of myristate.     

Evaluation of Antioxidant,Antibacterial and Enhancement of Antibiotic Action by Punica granatum Leaves Crude Extract and Enriched Fraction against Multidrug-Resistant Bacteria

The aim of this study was to evaluate the phytochemical composition, antioxidant, and antimicrobial potential of crude extract and fractions of Punica granatum leaves. The extract was produced by turbo extraction, after which hexanic, ethyl acetate, and aqueous fractions were obtained by partitioning. The chemical analyses were performed by thin layer chromatography and high-performance liquid chromatography, and the antioxidant activities were assayed by DPPH^. and ABTS^.+. Minimal inhibitory and bactericidal concentrations (MIC/MBC) were applied to twenty-two bacteria. Most strains susceptible to extract/fractions and resistant to antibiotics were selected, and ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with the ethyl acetate fraction (EAF) against multidrug-resistant strains in modulatory and checkboard models. The data from chromatographic analyses showed flavonoids and tannins in the extract, as well as the enrichment of EAF in phenols, mainly flavonoids. The flavonoids were connected to the electron transfer activity demonstrated in the DPPH^. and ABTS^.+ assays. Gram-positive strains are more susceptible to EAF. The subinhibitory concentrations of P. granatum enhanced the antimicrobial activity of the agents and reduced the EAF individual MIC, and the combination of EAF and antibiotics demonstrated a synergistic effect. These results present a promising approach for developing a therapy in which antioxidant extracts and fractions can be used in combination with antibiotics.     

Effect of diets rich in N-3 polyunsatured fatty acids on muscle lipids and fatty acids in Belgian Blue double-muscled young bulls

The present study was aimed at investigating the effect of duration and time of feeding n-3 fatty acids on the fatty acid composition of intramuscular fat and adipose tissue of bulls at slaughter. Four groups of bulls were given during three periods different diets, mainly differing in the presence of linseed as the predominant n-3 fatty acid source in the concentrate either or not in combination with grass (silage) as the roughage. The results show that the fatty acid composition of the feed during the earlier periods of life of the animal were important and influenced the final intramuscular fatty acid composition. Feeding n-3 PUFA during the phases before the finishing diet increased the long chain n-3 PUFA (C20:5n-3, C22:5n-3 and C22:6n-3) compared to animals which were fed only a C 18:3n-3 rich concentrate in the finishing period. The cis-9,trans-11CLA content was increased by feeding linseed in the fattening period and was mainly deposited in the triacylglycerol fraction of the intramuscular fat.     

Intrinsic Motivations Drive Learning of Eye Movements: An Experiment with Human Adults

Intrinsic motivations drive the acquisition of knowledge and skills on the basis of novel or surprising stimuli or the pleasure to learn new skills. In so doing, they are different from extrinsic motivations that are mainly linked to drives that promote survival and reproduction. Intrinsic motivations have been implicitly exploited in several psychological experiments but, due to the lack of proper paradigms, they are rarely a direct subject of investigation. This article investigates how different intrinsic motivation mechanisms can support the learning of visual skills, such as “foveate a particular object in space”, using a gaze contingency paradigm. In the experiment participants could freely foveate objects shown in a computer screen. Foveating each of two “button” pictures caused different effects: one caused the appearance of a simple image (blue rectangle) in unexpected positions, while the other evoked the appearance of an always-novel picture (objects or animals). The experiment studied how two possible intrinsic motivation mechanisms might guide learning to foveate one or the other button picture. One mechanism is based on the sudden, surprising appearance of a familiar image at unpredicted locations, and a second one is based on the content novelty of the images. The results show the comparative effectiveness of the mechanism based on image novelty, whereas they do not support the operation of the mechanism based on the surprising location of the image appearance. Interestingly, these results were also obtained with participants that, according to a post experiment questionnaire, had not understood the functions of the different buttons suggesting that novelty-based intrinsic motivation mechanisms might operate even at an unconscious level.     

Identification of Novel Loci Associated with Gastrointestinal Parasite Resistance in a Red Maasai x Dorper Backcross Population

Gastrointestinal (GI) parasitic infection is the main health constraint for small ruminant production, causing loss of weight and/or death. Red Maasai sheep have adapted to a tropical environment where extreme parasite exposure is a constant, especially with highly pathogenic Haemonchus contortus. This breed has been reported to be resistant to gastrointestinal parasite infection, hence it is considered an invaluable resource to study associations between host genetics and resistance. The aim of this study was to identify polymorphisms strongly associated with host resistance in a double backcross population derived from Red Maasai and Dorper sheep using a SNP-based GWAS analysis. The animals that were genotyped represented the most resistant and susceptible individuals based on the tails of phenotypic distribution (10% each) for average faecal egg counts (AVFEC). AVFEC, packed cell volume (AVPCV), and live weight (AVLWT) were adjusted for fixed effects and co-variables, and an association analysis was run using EMMAX. Revised significance levels were calculated using 100,000 permutation tests. The top five significant SNP markers with - log10 p-values >3.794 were observed on five different chromosomes for AVFEC, and BLUPPf90/PostGSf90 results confirmed EMMAX significant regions for this trait. One of these regions included a cluster of significant SNP on chromosome (Chr) 6 not in linkage disequilibrium to each other. This genomic location contains annotated genes involved in cytokine signalling, haemostasis and mucus biosynthesis. Only one association detected on Chr 7 was significant for both AVPCV and AVLWT. The results generated here reveal candidate immune variants for genes involved in differential response to infection and provide additional SNP marker information that has potential to aid selection of resistance to gastrointestinal parasites in sheep of a similar genetic background to the double backcross population.