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Paracellular permeability is increased by basal lipopolysaccharide in a primary culture of colonic epithelial cells; an effect prevented by an activator of Toll-like receptor-2.

Innate Immun. 2011;17(3):269-82

Authors: Hanson PJ, Moran AP, Butler K

Abstract
Lipopolysaccharide (LPS), which generally activates Toll-like receptor 4 (TLR4), is expressed on commensal colonic bacteria. In a number of tissues, LPS can act directly on epithelial cells to increase paracellular permeability. Such an effect in the colon would have an important impact on the understanding of normal homeostasis and of pathology. Our aim was to use a novel primary culture of colonic epithelial cells grown on Transwells to investigate whether LPS, or Pam(3)CSK( 4), an activator of TLR2, affected paracellular permeability. Consequently, [(14)C]-mannitol transfer and transepithelial electrical resistance (TEER) were measured. The preparation consisted primarily of cytokeratin-18 positive epithelial cells that produced superoxide, stained for mucus with periodic acid-Schiff reagent, exhibited alkaline phosphatase activity and expressed TLR2 and TLR4. Tight junctions and desmosomes were visible by transmission electron microscopy. Basally, but not apically, applied LPS from Escherichia coli increased the permeability to mannitol and to a 10-kDa dextran, and reduced TEER. The LPS from Helicobacter pylori increased paracellular permeability of gastric cells when applied either apically or basally, in contrast to colon cells, where this LPS was active only from the basal aspect. A pan-caspase inhibitor prevented the increase in caspase activity caused by basal E. coli LPS, and reduced the effects of LPS on paracellular permeability. Synthetic Pam(3)CSK(4) in the basal compartment prevented all effects of basal E. coli LPS. In conclusion, LPS applied to the base of the colonic epithelial cells increased paracellular permeability by a mechanism involving caspase activation, suggesting a process by which perturbation of the gut barrier could be exacerbated. Moreover, activation of TLR2 ameliorated such effects.

PMID: 20472611 [PubMed - indexed for MEDLINE]

Intravaginal administration of isosorbide mononitrate and misoprostol for cervical ripening and induction of labour: a randomized controlled trial.

Arch Gynecol Obstet. 2011 Jul;284(1):25-30

Authors: Abdellah MS, Hussien M, Aboalhassan A

Abstract
BACKGROUND: Labor induction in the presence of unfavorable cervix is a common indication for the use of prostaglandins. However, in the last years, there has been a considerable interest in the use of nitrous oxide donors for cervical ripening and labor induction.
OBJECTIVE: To evaluate the efficacy and safety of intravaginal administration of NOD isosorbide mononitrate (IMN) plus misoprostol versus misoprostol alone for cervical ripening and induction of labor.
SETTING: Department of Obstetrics and Gynecology, Woman's Health Center, Assiut University.
METHODS: Two hundred and ninety women scheduled for labor induction were recruited and assigned randomly to IMN or placebo followed by misoprostol 50 ?g. The efficacy of the medication was evaluated by predetermined outcome variables for cervical ripening and induction of labor and delivery.
RESULTS: The two groups were comparable with respect to age, parity, gestational age, indication for induction, and Bishop's score. Women receiving IMN plus misoprostol showed significant changes in the Bishop score 6 h after administration as compared to misoprostol plus placebo (8.57 ± 1.46 vs. 7.6 ± 1.39 h, P = 0.001), significantly shorter intervals from the beginning of the induction to the beginning of the active phase of labor (10.97 ± 2.87 vs. 13.91 ± 2.16 h, P = 0.0004) and from the beginning of induction to the time of delivery (19.56 ± 3.96 vs. 23 ± 2.62 P ? 0.001). No significant differences in the incidence of uterine hypersystole, tachysystole and hyperstimulation. Regarding headache, much more women suffer headache in the IMN group (51) with significant difference to placebo group (11).
CONCLUSIONS: Using a combination of IMN and misoprostol is more efficient than misoprostol alone in terms of fast cervical ripening and shortening of induction-labour interval.

PMID: 20582425 [PubMed - indexed for MEDLINE]

Caloric restriction reduces IgA levels and modifies cytokine mRNA expression in mouse small intestine.

J Nutr Biochem. 2011 Jun;22(6):560-6

Authors: Lara-Padilla E, Campos-Rodríguez R, Jarillo-Luna A, Reyna-Garfias H, Rivera-Aguilar V, Miliar A, Berral de la Rosa FJ, Navas P, López-Lluch G

Abstract
The aim of this study was to determine the effect of caloric restriction (CR) in mouse small intestine on the production and secretion of immunoglobulin (Ig) A, the population of lymphocytes in the lamina propria, and the expression of cytokines that mediate and regulate innate and adaptive immunity. One group of young Balb/c mice was fed ad libitum, while the CR group was fed ad libitum and fasted on alternate days. When mice were six months old, IgA levels in the proximal small intestine were quantified by enzyme-linked immunosorbent assay, while the number of IgA containing cells, CD4(+) T cells and CD8(+) T cells in the duodenal mucosa was determined by immunohistochemistry. Furthermore, the expression of several intestinal cytokines, the genes for ?-chain IgA, and the polymeric Ig receptor (pIgR) were analyzed by real-time polymerase chain reaction. CR decreased the levels of IgA in the intestine, apparently a consequence of a reduced number of IgA(+) cells in the lamina propria that decrease the production and secretion of this Ig, and a reduced secretion of S-IgA into the bile, which in turn discharges into the proximal intestine. Contrarily, CR increased the expression of genes for ?-chain IgA, and the pIgR, indicating that transport of IgA was not a key factor in the decrease of this Ig. Additionally, CR modified the expression of genes for tumor necrosis factor-?, interferon-?, tumor growth factor-?, interleukin (IL)-2 and IL-10, all of which regulate the synthesis of IgA and pIgR, the inflammatory response, and the immune response in the intestine.

PMID: 20951020 [PubMed - indexed for MEDLINE]

Cloning and characterization of a wheat neutral ceramidase gene Ta-CDase.

Mol Biol Rep. 2011 Jun;38(5):3447-54

Authors: Yu X, Wang X, Huang X, Buchenauer H, Han Q, Guo J, Zhao J, Qu Z, Huang L, Kang Z

Abstract
Ceramidases are key enzymes in the regulation of the cellular levels of ceramide, sphingosine and sphingosine-1-phosphate. This study first reports on the molecular cloning, sequencing and expression profile of the gene encoding the wheat neutral ceramidase designated as Ta-CDase. A full length wheat Ta-CDase gene is obtained by rapid amplification of cDNA ends (RACE) based on the sequence of the WSRC36 fragment from an incompatible suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The open reading frame (ORF) of 2,839 nucleotides encodes a polypeptide of 785 amino acids with a predicted isoelectric point (pI) of 6.398. The protein conserved domain search indicates that the polypeptide contains the signature of ceramidase, signal peptide sequence and transmembrane region. A phylogenetic analysis reveals that a high degree of relatedness exists among wheat Ta-CDase and ceramidases from other plant species at the amino acid level, while its relationship to that of animals and pathogens is more distant. The expression profile of the Ta-CDase shows a very strong expression of transcripts only at 48 h post inoculation (hpi), while expression level is low at other time points. Southern blot analyses showed that Ta-CDase is a multi-copy gene and located on wheat chromosome 4D and 5A.

PMID: 21088901 [PubMed - indexed for MEDLINE]

Intramyocardial adrenergic activation in chagasic cardiomyopathy and coronary artery disease.

Arq Bras Cardiol. 2011 Feb;96(2):99-106

Authors: Nastari L, Ramires FJ, Salemi VM, Ianni BM, Fernandes F, Strunz CM, Arteaga E, Mady C

Abstract
BACKGROUND: Myocardial norepinephrine is altered in left ventricular impairment. In patients with Chagas' cardiomyopathy (CC), this issue has not been addressed.
OBJECTIVE: To determine the level of myocardial norepinephrine in patients with CC and compare it in patients with coronary artery disease, and to relate myocardial norepinephrine to left ventricular ejection fraction (LVEF).
METHODS: We studied 39 patients with CC, divided into group 1: 21 individuals with normal LVEF and group 2: 18 individuals with decreased LVEF. Seventeen patients with coronary artery disease were divided into group 3: 12 individuals with normal LVEF and group 4: 5 individuals with decreased LVEF. Two-dimensional echocardiography was used to measure LVEF. Myocardial norepinephrine was determined by high-performance liquid chromatography.
RESULTS: Myocardial norepinephrine in CC with and without ventricular dysfunction was 1.3±1.3 and 6.1±4.2 pg/?g noncollagen protein, respectively (p<0.0001); in coronary artery disease with and without ventricular dysfunction, it was 3.3±3.0 and 9.8±4.2 pg/?g noncollagen protein, respectively (p<0.0001). A positive correlation was found between LVEF and myocardial norepinephrine concentration in the patients with Chagas' cardiomyopathy (p<0.01, r = 0.57) and also in those with coronary artery disease (p<0.01, r=0.69). A significant difference was demonstrated between norepinephrine concentrations in patients with normal LVEF (groups 1 and 3; p = 0.0182), but no difference was found in patients with decreased LVEF (groups 2 and 4; p = 0.1467).
CONCLUSION: In patients with Chagas' cardiomyopathy and normal global ejection fraction there is an early cardiac denervation, when compared to coronary artery disease patients.

PMID: 21180891 [PubMed - indexed for MEDLINE]

Preparation of the monomers of gingerols and 6-shogaol by flash high speed counter-current chromatography.

J Chromatogr A. 2011 Sep 9;1218(36):6187-90

Authors: Qiao Q, Du Q

Abstract
The flash high speed counter-current chromatographic (FHSCCC) separation of gingerols and 6-shogaol was performed on a HSCCC instrument equipped with a 1200-ml column (5 mm tubing i.d.) at a flow rate of 25 ml/min. The performance met the FHSCCC feature that the flow rate of mobile phase (ml) is equal to or greater than the square of the diameter of the column tubing (mm). The separation employed the upper phase of stationary phase of the n-hexane-ethyl acetate-methanol-water (3:2:2:3, v/v) as the stationary phase. A stepwise elution was performed by eluting with the lower phase of n-hexane-ethyl acetate-methanol-water (3:2:2:3, v/v) for first 90 min and the lower phase of the n-hexane-ethyl acetate-methanol-water (3:2:6:5, v/v) for the second 90 min. In each separation 5 g of the ethyl acetate extract of rhizomes of ginger was loaded, yielding 1.96 g of 6-gingerol (98.3%), 0.33 g of 8-gingerol (97.8%), 0.64 g of 6-shogaol (98.8%) and 0.57 g of 10-gingerol (98.2%). The separation can be expected to scale up to industrial separation.

PMID: 21195411 [PubMed - indexed for MEDLINE]

Synthesis of 7,8?-dihydro-14?-deoxyecdysteroids.

Steroids. 2011 May;76(6):603-6

Authors: Savchenko RG, Urasaeva YR, Galyautdinov IV, Afonkina SR, Khalilov LM, Dolgushin FM, Odinokov VN

Abstract
A Pd-C-catalyzed hydrogenation in methanol and in the presence of sodium methylate is a simple, convenient and high yielding reduction method to convert the 7,14-dien-6-one ecdysteroids to their corresponding 7,8?-dihydro-14?-deoxyecdysteroids.

PMID: 21356225 [PubMed - indexed for MEDLINE]

The tramadol metabolite O-desmethyl tramadol inhibits substance P-receptor functions expressed in Xenopus oocytes.

J Pharmacol Sci. 2011;115(3):421-4

Authors: Minami K, Yokoyama T, Ogata J, Uezono Y

Abstract
Tramadol has been widely used as analgesic. O-Desmethyl tramadol (ODT) is one of the main metabolites of tramadol, having much greater analgesic potency than tramadol itself. Substance P receptors (SPR) are well known to modulate nociceptive transmission within the spinal cord. In this study, we investigated the effects of ODT on SPR expressed in Xenopus oocytes by examining SP-induced Ca(2+)-activated Cl(-) currents. ODT inhibited the SPR-induced Cl(-) currents at pharmacologically relevant concentrations. The protein kinase C (PKC) inhibitor bisindolylmaleimide I did not abolish the inhibitory effects of ODT on SP-induced Ca(2+)-activated Cl(-) currents. The results suggest that the tramadol metabolite ODT inhibits the SPR functions, which may be independent of activation of PKC-mediated pathways.

PMID: 21372504 [PubMed - indexed for MEDLINE]

Tonic inhibition of allergic itch signaling by the descending noradrenergic system in mice.

J Pharmacol Sci. 2011;115(3):417-20

Authors: Gotoh Y, Omori Y, Andoh T, Kuraishi Y

Abstract
We investigated whether the descending noradrenergic system regulates allergic itch. Mosquito allergy of the hind paw elicited biting, an itch-related response, in sensitized mice. The biting was inhibited by intrathecal clonidine and reversed by yohimbine, an ?(2)-adrenoceptor antagonist. The biting was increased by intrathecal pretreatment with the catecholaminergic neurotoxin 6-hydroxydopamine and the ?-adrenoceptor antagonist phentolamine but not the serotonergic neurotoxin 5,7-dihydroxytryptamine. We propose that ?(2)-adrenoceptors are involved in the inhibition of allergic itch in the spinal cord and that the descending noradrenergic system exerts a tonic inhibition on the itch signaling. The serotonergic system may not be involved.

PMID: 21372505 [PubMed - indexed for MEDLINE]

Site-specific modification of recombinant proteins: a novel platform for modifying glycoproteins expressed in E. coli.

Bioconjug Chem. 2011 May 18;22(5):903-12

Authors: Henderson GE, Isett KD, Gerngross TU

Abstract
The site-specific modification of proteins is expected to be an important capability for the synthesis of bioconjugates in the future. However, the traditional repertoire of reactions available for the direct modification of proteins suffers from lack of specificity, necessitating costly downstream processing to isolate the specific species of interest. (1) Here, we use a well-established, glycan-specific chemistry to PEGylate model glycoproteins, each containing a unique reactive GalNAc attached to a specifically engineered threonine residue. By engineering E. coli to execute the initial steps of human, mucin-type O-glycosylation, we were able to obtain homogeneous site-specifically modified glycoproteins with fully human glycan linkages. Two mucin-based reporters as well as several fusion proteins containing eight-amino-acid GalNAc-T recognition sequences were glycosylated in this engineered glycocompetent strain of E. coli. The use of one sequence in particular, PPPTSGPT, resulted in site-specific glycan occupancy of approximately 69% at the engineered threonine. The GalNAc present on the purified glycoprotein was oxidized by galactose oxidase and then coupled to hydroxylamine functionalized 20 kDa PEG in the presence of aniline. The glycoprotein could be converted to the PEGylated product at approximately 85% yield and >98% purity as determined by comparison to the products of control reactions.

PMID: 21395336 [PubMed - indexed for MEDLINE]

Synthesis and in vitro insulin-mimetic activities of zinc(ii) complexes of ethyl 2,5-dihydro-4-hydroxy-5-oxo-1H-pyrrole-3-carboxylates.

Metallomics. 2011 Jul;3(7):675-9

Authors: Kawarada H, Yoshikawa Y, Yasui H, Kuwahara S, Habata Y, Saito R

Abstract
Several Zn(II) complexes (2a-e) of 1-arylmethyl-2,5-dihydro-4-hydroxy-5-oxo-1H-pyrrole-3-carboxylates (1a-e), known drug candidates for diabetic complications, were synthesized and proved to have in vitro insulin-mimetic activities, suggesting that these complexes are potential chemotherapeutics that are effective against both diabetes and diabetic complications.

PMID: 21423934 [PubMed - indexed for MEDLINE]

Targeted albumin-based nanoparticles for delivery of amphipathic drugs.

Bioconjug Chem. 2011 May 18;22(5):870-8

Authors: Xu R, Fisher M, Juliano RL

Abstract
We report the preparation and physical and biological characterization of human serum albumin-based micelles of approximately 30 nm diameter for the delivery of amphipathic drugs, represented by doxorubicin. The micelles were surface conjugated with cyclic RGD peptides to guide selective delivery to cells expressing the ?(v)?(3) integrin. Multiple poly(ethylene glycol)s (PEGs) with molecular weight of 3400 Da were used to form a hydrophilic outer layer, with the inner core formed by albumin conjugated with doxorubicin via disulfide bonds. Additional doxorubicin was physically adsorbed into this core to attain a high drug loading capacity, where each albumin was associated with about 50 doxorubicin molecules. The formed micelles were stable in serum but continuously released doxorubicin when incubated with free thiols at concentrations mimicking the intracellular environment. When incubated with human melanoma cells (M21+) that express the ?(v)?(3) integrin, higher uptake and longer retention of doxorubicin was observed with the RGD-targeted micelles than in the case of untargeted control micelles or free doxorubicin. Consequently, the RGD-targeted micelles manifested cytotoxicity at lower doses of drug than control micelles or free drug.

PMID: 21452893 [PubMed - indexed for MEDLINE]

Surfactant blocks lipopolysaccharide signaling by inhibiting both NF?B and PARP activation in experimental ARDS.

Mol Cell Biochem. 2011 Aug;354(1-2):113-22

Authors: Mittal N, Sanyal SN

Abstract
Surfactant plays an important role in lung homeostasis and is also involved in maintaining innate immunity within the lung. Lipopolysaccharide (LPS) is known to elicit acute proinflammatory responses in lung diseases such as acute respiratory distress syndrome and is responsible for the expression of the inducible isoform of nitric oxide synthase (iNOS). Because cells are exposed to low pH within the microenvironment of inflammatory lesions, the potential role of low environmental pH on iNOS expression was investigated. Low environmental pH-induced iNOS gene transcription involved the activation of nuclear factor-?B (NF-?B) transcription factor and I?B kinases. Here, we find that exposure of cells to low environmental pH increased both nitrite accumulation and activation of NF-?B-signaling pathway by Western blot and immunohistochemistry. In addition, treatment with surfactant prevents NF-?B translocation to the nucleus by preventing phosphorylation of I?B?, and its subsequent degradation by IKK?, and canceling low pH-induced nitrite accumulation. Surfactant also inhibited the LPS-induced PARP activation. Therefore, surfactant may regulate lung homeostasis by neutralizing acidic microenvironment in inflammatory lesions that leads to the upregulation of iNOS activity through the activation of NF-?B pathway and by PARP activation.

PMID: 21461608 [PubMed - indexed for MEDLINE]

Enhanced brain stem 5HT?A receptor function under neonatal hypoxic insult: role of glucose, oxygen, and epinephrine resuscitation.

Mol Cell Biochem. 2011 Aug;354(1-2):151-60

Authors: Anju TR, Korah PK, Jayanarayanan S, Paulose CS

Abstract
Molecular processes regulating brain stem serotonergic receptors play an important role in the control of respiration. We evaluated 5-HT(2A) receptor alterations in the brain stem of neonatal rats exposed to hypoxic insult and the effect of glucose, oxygen, and epinephrine resuscitation in ameliorating these alterations. Hypoxic stress increased the total 5-HT and 5-HT(2A) receptor number along with an up regulation of 5-HT Transporter and 5-HT(2A) receptor gene in the brain stem of neonates. These serotonergic alterations were reversed by glucose supplementation alone and along with oxygen to hypoxic neonates. The enhanced brain stem 5-HT(2A) receptors act as a modulator of ventilatory response to hypoxia, which can in turn result in pulmonary vasoconstriction and cognitive dysfunction. The adverse effects of 100% oxygenation and epinephrine administration to hypoxic neonates were also reported. This has immense clinical significance in neonatal care.

PMID: 21484469 [PubMed - indexed for MEDLINE]

Damaged peroxisomes are subject to rapid autophagic degradation in the yeast Hansenula polymorpha.

Autophagy. 2011 Aug 1;7(8):863-72

Authors: van Zutphen T, Veenhuis M, van der Klei IJ

Abstract
Evidence is accumulating that damaged components of eukaryotic cells are removed by autophagic degradation (e.g., mitophagy). Here we show that peroxisomes that are damaged by the abrupt removal of the membrane protein Pex3 are massively and rapidly degraded even when the cells are placed at peroxisome-inducing conditions and hence need the organelles for growth. Pex3 degradation was induced by a temperature shift using Hansenula polymorpha pex3? cells producing a Pex3 fusion protein containing an N-terminal temperature sensitive degron sequence. The massive peroxisome degradation process, associated with Pex3 degradation, showed properties of both micro- and macropexophagy and was dependent on Atg1 and Ypt7. This mode of peroxisome degradation is of physiological significance as it was also observed at conditions that excessive ROS is formed from peroxisome metabolism, i.e., when methanol-grown wild-type cells are exposed to methanol excess conditions.

PMID: 21490428 [PubMed - indexed for MEDLINE]

Intermittent counter-current extraction--effect of the key operating parameters on selectivity and throughput.

J Chromatogr A. 2011 Sep 9;1218(36):6072-8

Authors: Hewitson P, Ignatova S, Sutherland I

Abstract
Intermittent counter-current extraction (ICcE) has proved itself as a method for splitting compounds into streams and/or concentrating compounds in the column. In this paper a model mixture sample based on a modified GUESSmix (containing salicin, caffeine, aspirin, coumarin, salicylic acid, carvone, ionone and biphenyl) was separated into two eluant streams across a range of HEMWat phase system polarities from the polar system 11 through to non-polar system 23. ICcE could provide throughput of over 1 kg/day with this model sample, at the preparative scale, Changing the time cycle to adjust where the sample mixture is split into two streams was demonstrated. It is established that for the continuous running of ICcE, on a conventional twin bobbin counter-current chromatograph instrument, it is necessary to adjust the dead volumes of the flying leads to maintain similar phase retention in each column so the instrument does not become hydrodynamically and mechanically unbalanced due to the difference in densities between the upper and lower phases.

PMID: 21571282 [PubMed - indexed for MEDLINE]

Low-dose adrenaline, promethazine, and hydrocortisone in the prevention of acute adverse reactions to antivenom following snakebite: a randomised, double-blind, placebo-controlled trial.

PLoS Med. 2011 May;8(5):e1000435

Authors: de Silva HA, Pathmeswaran A, Ranasinha CD, Jayamanne S, Samarakoon SB, Hittharage A, Kalupahana R, Ratnatilaka GA, Uluwatthage W, Aronson JK, Armitage JM, Lalloo DG, de Silva HJ

Abstract
BACKGROUND: Envenoming from snakebites is most effectively treated by antivenom. However, the antivenom available in South Asian countries commonly causes acute allergic reactions, anaphylactic reactions being particularly serious. We investigated whether adrenaline, promethazine, and hydrocortisone prevent such reactions in secondary referral hospitals in Sri Lanka by conducting a randomised, double-blind placebo-controlled trial.
METHODS AND FINDINGS: In total, 1,007 patients were randomized, using a 2 × 2 × 2 factorial design, in a double-blind, placebo-controlled trial of adrenaline (0.25 ml of a 1?1,000 solution subcutaneously), promethazine (25 mg intravenously), and hydrocortisone (200 mg intravenously), each alone and in all possible combinations. The interventions, or matching placebo, were given immediately before infusion of antivenom. Patients were monitored for mild, moderate, or severe adverse reactions for at least 96 h. The prespecified primary end point was the effect of the interventions on the incidence of severe reactions up to and including 48 h after antivenom administration. In total, 752 (75%) patients had acute reactions to antivenom: 9% mild, 48% moderate, and 43% severe; 89% of the reactions occurred within 1 h; and 40% of all patients were given rescue medication (adrenaline, promethazine, and hydrocortisone) during the first hour. Compared with placebo, adrenaline significantly reduced severe reactions to antivenom by 43% (95% CI 25-67) at 1 h and by 38% (95% CI 26-49) up to and including 48 h after antivenom administration; hydrocortisone and promethazine did not. Adding hydrocortisone negated the benefit of adrenaline.
CONCLUSIONS: Pretreatment with low-dose adrenaline was safe and reduced the risk of acute severe reactions to snake antivenom. This may be of particular importance in countries where adverse reactions to antivenom are common, although the need to improve the quality of available antivenom cannot be overemphasized.

PMID: 21572992 [PubMed - indexed for MEDLINE]

An effective technique for the storage of short lived radioactive gaseous waste.

Appl Radiat Isot. 2011 Sep;69(9):1185-8

Authors: Schweiger L

Abstract
An effective technique is described to deal with volatile, short lived radioactive waste generated as a result of the routinely produced positron emission tomography (PET) radiopharmaceutical 2-deoxy-2-[(18)F]fluoro-D-glucose (FDG). All radioactive gases and aerosols created during the synthesis are collected and stored safely in commercially available TEDLAR gas sampling bags. Once these collected PET by-products decay, the TEDLAR gas bags can be easily emptied and reused. This improved technique is effective, safe, reliable and economical.

PMID: 21592805 [PubMed - indexed for MEDLINE]

Real-time quantification of microscale bioadhesion events in situ using imaging surface plasmon resonance (iSPR).

ACS Appl Mater Interfaces. 2011 Jun;3(6):2085-91

Authors: Aldred N, Ekblad T, Andersson O, Liedberg B, Clare AS

Abstract
From macro- to nanoscales, adhesion phenomena are all-pervasive in nature yet remain poorly understood. In recent years, studies of biological adhesion mechanisms, terrestrial and marine, have provided inspiration for "biomimetic" adhesion strategies and important insights for the development of fouling-resistant materials. Although the focus of most contemporary bioadhesion research is on large organisms such as marine mussels, insects and geckos, adhesion events on the micro/nanoscale are critical to our understanding of important underlying mechanisms. Observing and quantifying adhesion at this scale is particularly relevant for the development of biomedical implants and in the prevention of marine biofouling. However, such characterization has so far been restricted by insufficient quantities of material for biochemical analysis and the limitations of contemporary imaging techniques. Here, we introduce a recently developed optical method that allows precise determination of adhesive deposition by microscale organisms in situ and in real time; a capability not before demonstrated. In this extended study we used the cypris larvae of barnacles and a combination of conventional and imaging surface plasmon resonance techniques to observe and quantify adhesive deposition onto a range of model surfaces (CH(3)-, COOH-, NH(3)-, and mPEG-terminated SAMs and a PEGMA/HEMA hydrogel). We then correlated this deposition to passive adsorption of a putatively adhesive protein from barnacles. In this way, we were able to rank surfaces in order of effectiveness for preventing barnacle cyprid exploration and demonstrate the importance of observing the natural process of adhesion, rather than predicting surface effects from a model system. As well as contributing fundamentally to the knowledge on the adhesion and adhesives of barnacle larvae, a potential target for future biomimetic glues, this method also provides a versatile technique for laboratory testing of fouling-resistant chemistries.

PMID: 21595456 [PubMed - indexed for MEDLINE]

Dysregulation of IFN system can lead to poor response to pegylated interferon and ribavirin therapy in chronic hepatitis C.

PLoS One. 2011;6(5):e19799

Authors: Onomoto K, Morimoto S, Kawaguchi T, Toyoda H, Tanaka M, Kuroda M, Uno K, Kumada T, Matsuda F, Shimotohno K, Fujita T, Murakami Y

Abstract
BACKGROUND: Despite being expensive, the standard combination of pegylated interferon (Peg-IFN)-? and ribavirin used to treat chronic hepatitis C (CH) results in a moderate clearance rate and a plethora of side effects. This makes it necessary to predict patient outcome so as to improve the accuracy of treatment. Although the antiviral mechanism of genetically altered IL28B is unknown, IL28B polymorphism is considered a good predictor of IFN combination treatment outcome.
METHODOLOGY: Using microarray, we quantified the expression profile of 237 IFN related genes in 87 CH liver biopsy specimens to clarify the relationship between IFN pathway and viral elimination, and to predict patients' clinical outcome. In 72 out of 87 patients we also analyzed IL28B polymorphism (rs8099917).
PRINCIPAL FINDINGS: Five IFN related-genes (IFI27, IFI 44, ISG15, MX1, and OAS1) had expression levels significantly higher in nonresponders (NR) than in normal liver (NL) and sustained virological responders (SVR); this high expression was also frequently seen in cases with the minor (TG or GG) IL28B genotype. The expression pattern of 31 IFN related-genes also differed significantly between NR and NL. We predicted drug response in NR with 86.1% accuracy by diagonal linear discriminant analysis (DLDA).
CONCLUSION: IFN system dysregulation before treatment was associated with poor IFN therapy response. Determining IFN related-gene expression pattern based on patients' response to combination therapy, allowed us to predict drug response with high accuracy. This method can be applied to establishing novel antiviral therapies and strategies for patients using a more individual approach.

PMID: 21603632 [PubMed - indexed for MEDLINE]

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